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Sample records for mutant pisum sativum

  1. Development of the alt Mutant of Pisum sativum L. 1

    PubMed Central

    Guo, Wei Wen; Proebsting, William M.; Potter, Sandra W.; Daley, Larry S.; Potter, John R.

    1987-01-01

    The alt (albina-terminalis) mutant of Pisum sativum L. germinates normally, produces several nodes, and then above a sharp transition produces 2 to 3 bleached nodes, ceases growth, and eventually dies. Green nodes have normal chlorophyll content, absorption spectra, photosynthetic rates, and ultrastructure. In bleaching tissues, the chloroplasts degenerate rapidly, followed by extensive disruption and loss of the remaining cytoplasm and organelles. Application of tissue extracts of normal genotypes of pea, corn, and bean stimulates apical development of alt. The resulting tissues have essentially normal structure and function. Application of thiamine, thiamine monophosphate, and thiamine pyrophosphate also stimulate normal apical development at concentrations of 1 micromolar and above. Partial characterization of the stimulus from pea seed extracts is consistent with thiamine as the active factor. Images Fig. 4 Fig. 6 Fig. 8 PMID:16665809

  2. Ethylene Inhibitors Restore Nodulation to sym 5 Mutants of Pisum sativum L. cv Sparkle 12

    PubMed Central

    Fearn, Jeffrey C.; LaRue, Thomas A.

    1991-01-01

    The sym 5 mutants of pea, Pisum sativum L. cv Sparkle, do not differ in growth habit from their normal parent and nodulate poorly at a root temperature of 20°C. If inhibitors of ethylene formation or action (Co2+, aminoethoxyvinylglycine, or Ag+) are added to the substrate, nodulation of the sym 5 mutants is increased. Similar treatments of four other mutant sym lines do not restore nodulation. When Ag+ is added to the substrate from 4 days before to 4 days after inoculation with rhizobia, nodulation of sym 5 mutants is increased. The roots of the mutant need only be exposed to Ag+ for 4 hours to significantly increase nodule numbers. The content of free 1-aminocyclopropane-1-carboxylic acid and the production of ethylene in the lateral roots of sym 5 mutants do not differ from Sparkle. PMID:16668158

  3. Stomatal responses of Argenteum - a mutant of Pisum sativum L. with readily detachable leaf epidermis.

    PubMed

    Jewer, P C; Incoll, L D; Shaw, J

    1982-07-01

    Epidermis is easily detached from both adaxial and abaxial surfaces of leaf four of the Argenteum mutant of Pisum sativum L. The isolated epidermis has stomata with large, easily-measured pores. Hairs and glands are absent. The density of stomata is high and contamination by mesophyll cells is low. In the light and in CO2-free air, stomata in isolated adaxial epidermis of Argenteum mutant opened maximally after 4 h incubation at 25°C. The response of stomata to light was dependent on the concentration of KCl in the incubation medium and was maximal at 50 mol m(-3) KCl. Stomata did not respond to exogenous kinetin, but apertures were reduced by incubation of epidermis on solutions containing between 10(-5) and 10(-1) mol m(-3) abscisic acid (ABA). The responses of stomata of Argenteum mutant to light, exogenous KCl, ABA and kinetin were comparable with those described previously for stomata in isolated epidermis of Commelina communis. A method for preparing viable protoplasts of guard cells from isolated epidermis of Argenteum mutant is described. The response of guard cell protoplasts to light, exogenous KCl, ABA and kinetin were similar to those of stomata in isolated epidermis except that the increase in volume of the protoplasts in response to light was maximal at a lower concentration of KCl (10 mol m(-3)) and that protoplasts responded more rapidly to light than stomata in isolated epidermis. The protoplasts did not respond to exogenous kinetin, but when incubated for 1 h in the light and in CO2-free air on a solution containing 10(-3) mol m(-3) ABA, they decreased in volume by 30%. The advantages of using epidermis from Argenteum mutant for experiments on stomatal movements are discussed.

  4. Growth, seed development and genetic analysis in wild type and Def mutant of Pisum sativum L

    PubMed Central

    2011-01-01

    Background The def mutant pea (Pisum sativum L) showed non-abscission of seeds from the funicule. Here we present data on seed development and growth pattern and their relationship in predicting this particular trait in wild type and mutant lines as well as the inheritance pattern of the def allele in F2 and F3 populations. Findings Pod length and seed fresh weight increase with fruit maturity and this may affect the abscission event in pea seeds. However, the seed position in either the distal and proximal ends of the pod did not show any difference. The growth factors of seed fresh weight (FW), width of funicles (WFN), seed width (SW) and seed height (SH) were highly correlated and their relationships were determined in both wild type and def mutant peas. The coefficient of determination R2 values for the relationship between WFN and FW, SW and SH and their various interactions were higher for the def dwarf type. Stepwise multiple regression analysis showed that variation of WFN was associated with SH and SW. Pearson's chi square analysis revealed that the inheritance and segregation of the Def locus in 3:1 ratio was significant in two F2 populations. Structural analysis of the F3 population was used to confirm the inheritance status of the Def locus in F2 heterozygote plants. Conclusions This study investigated the inheritance of the presence or absence of the Def allele, controlling the presence of an abscission zone (AZ) or an abscission-less zone (ALZ) forming in wild type and mutant lines respectively. The single major gene (Def) controlling this phenotype was monogenic and def mutants were characterized and controlled by the homozygous recessive def allele that showed no palisade layers in the hilum region of the seed coat. PMID:22078070

  5. Sink to Source Transition in Tendrils of a Semileafless Mutant, Pisum sativum cv Curly 1

    PubMed Central

    Côté, Richard; Gerrath, Jean M.; Peterson, Carol A.; Grodzinski, Bernard

    1992-01-01

    Sink to source transition parallels loss of thigmotropic capacity in tendrils of a semileafless mutant, Pisum sativum cv Curly. Macroscopic tendril development is subdivided based on thigmotropic capacity. Stage I is the elongation stage and, although the rate of photosynthesis is similar to that of stage II and III tendrils, dark respiration rates are higher in stage I. During stage II, tendrils are thigmotropic and act as a sink. Even though stage II tendrils have CO2 exchange characteristics similar to those of stage III tendrils, which are coiled, our fluorescein, 14C-partitioning, and 11C-translocation experiments suggest that stage I and II tendrils do not export carbon. Only stage III tendrils act as sources of newly fixed carbon. Export from them is blocked by cold, heat girdling of the petiole, or anoxia treatment of the tendrils. A late stage II tendril complex, in which coiling is occurring, may be exporting photoassimilates; however, this phenomenon can be attributed to the fact that the pea leaf is a compound structure and there may be one or more stage III tendrils, no longer thigmotropic, within the tendril complex. Photosynthetic maturity in pea tendrils occurs at stage III and is characterized by the ability of these tendrils to export photoassimilates. Images Figure 1 Figure 2 Figure 3 PMID:16653179

  6. The cadmium-tolerant pea (Pisum sativum L.) mutant SGECdt is more sensitive to mercury: assessing plant water relations

    PubMed Central

    Belimov, Andrey A.; Dodd, Ian C.; Safronova, Vera I.; Malkov, Nikita V.; Davies, William J.; Tikhonovich, Igor A.

    2015-01-01

    Heavy metals have multiple effects on plant growth and physiology, including perturbation of plant water status. These effects were assessed by exposing the unique Cd-tolerant and Cd-accumulating pea (Pisum sativum L.) mutant SGECdt and its wild-type (WT) line SGE to either cadmium (1, 4 μM CdCl2) or mercury (0.5, 1, 2 μM HgCl2) in hydroponic culture for 12 days. When exposed to Cd, SGECdt accumulated more Cd in roots, xylem sap, and shoot, and had considerably more biomass than WT plants. WT plants lost circa 0.2 MPa turgor when grown in 4 μM CdCl2, despite massive decreases in whole-plant transpiration rate and stomatal conductance. In contrast, root Hg accumulation was similar in both genotypes, but WT plants accumulated more Hg in leaves and had a higher stomatal conductance, and root and shoot biomass compared with SGECdt. Shoot excision resulted in greater root-pressure induced xylem exudation of SGECdt in the absence of Cd or Hg and following Cd exposure, whereas the opposite response or no genotypic differences occurred following Hg exposure. Exposing plants that had not been treated with metal to 50 μM CdCl2 for 1h increased root xylem exudation of WT, whereas 50 μM HgCl2 inhibited and eliminated genotypic differences in root xylem exudation, suggesting differences between WT and SGECdt plants in aquaporin function. Thus, root water transport might be involved in mechanisms of increased tolerance and accumulation of Cd in the SGECdt mutant. However, the lack of cross-tolerance to Cd and Hg stress in the mutant indicates metal-specific mechanisms related to plant adaptation. PMID:25694548

  7. Ultrastructural organization of chloroplast membranes in mutants of Pisum sativum L. with impaired activity in the photosystems.

    PubMed

    Popov, V I; Matorin, D N; Gostimsky, S V; Tageeva, S V; Allakhverdov, B L

    1981-05-01

    The ultrastructural organization and the photosynthesis reactions of chloroplast membranes were studied in three lethal mutants of Pisum sativum, Chl-1, Chl-19 and Chl-5, all lacking the capacity to evolve oxygen. The rates of 2,6-dichloroindophenol reduction, delayed fluorescence and electron-spin-resonance signal 1 indicate that Chl-1 and Chl-19 have an impaired activity in photosystem II (PS II), while in Chl-5 the electron transport is blocked between PS I and the reactions of CO2 fixation. Ultrathin sectioning demonstrates the presence of giant grana in the chloroplasts of Chl-1 and Chl-19, while the chloroplast structure of the Chl-5 is very similar to that of the wild-type. The grana of the Chl-19 mutant contain large multilamellar regions of tightly packed membranes. When the chloroplast membranes were studied by freeze-fracture, the exoplasmic and protoplasmic fracture faces (EF and PF, respectively) in both stacked and unstacked membranes were found to show large differences in particle concentrations and relative population area (per μm(2)), and also in particle size distribution, between all mutant chloroplast membranes and the wild-type. A close correlation between increasing kmt (ratio of particle concentrations on PF/EF) and PS II activity was observed. The differences in particle concentrations on both fracture faces in different regions of the intact chloroplast membranes of the wild-type are the consequence of a rearrangement of existing membrane components by lateral particle movements since quantitative measurements demonstrate almost complete conservation of intramembrane particles in number and size during the stacking of stroma thylakoid membranes. The results indicating particle movements strongly support the concept that the chloroplast membranes have a highly dynamic structure.

  8. Immunolocalization of pectic polysaccharides during abscission in pea seeds (Pisum sativum L.) and in abscission less def pea mutant seeds.

    PubMed

    Lee, YeonKyeong; Ayeh, Kwadwo Owusu; Ambrose, Mike; Hvoslef-Eide, Anne Kathrine

    2016-08-31

    In pea seeds (Pisum sativum L.), the presence of the Def locus determines abscission event between its funicle and the seed coat. Cell wall remodeling is a necessary condition for abscission of pea seed. The changes in cell wall components in wild type (WT) pea seed with Def loci showing seed abscission and in abscission less def mutant peas were studied to identify the factors determining abscission and non-abscission event. Changes in pectic polysaccharides components were investigated in WT and def mutant pea seeds using immunolabeling techniques. Pectic monoclonal antibodies (1 → 4)-β-D-galactan (LM5), (1 → 5)-α-L-arabinan(LM6), partially de-methyl esterified homogalacturonan (HG) (JIM5) and methyl esterified HG (JIM7) were used for this study. Prior to abscission zone (AZ) development, galactan and arabinan reduced in the predestined AZ of the pea seed and disappeared during the abscission process. The AZ cells had partially de-methyl esterified HG while other areas had highly methyl esterified HG. A strong JIM5 labeling in the def mutant may be related to cell wall rigidity in the mature def mutants. In addition, the appearance of pectic epitopes in two F3 populations resulting from cross between WT and def mutant parents was studied. As a result, we identified that homozygous dominant lines (Def/Def) showing abscission and homozygous recessive lines (def/def) showing non-abscission had similar immunolabeling pattern to their parents. However, the heterogeneous lines (Def/def) showed various immunolabeling pattern and the segregation pattern of the Def locus. Through the study of the complexity and variability of pectins in plant cell walls as well as understanding the segregation patterns of the Def locus using immunolabeling techniques, we conclude that cell wall remodeling occurs in the abscission process and de-methyl esterification may play a role in the non-abscission event in def mutant. Overall, this study contributes new insights into

  9. The cadmium-tolerant pea (Pisum sativum L.) mutant SGECdt is more sensitive to mercury: assessing plant water relations.

    PubMed

    Belimov, Andrey A; Dodd, Ian C; Safronova, Vera I; Malkov, Nikita V; Davies, William J; Tikhonovich, Igor A

    2015-04-01

    Heavy metals have multiple effects on plant growth and physiology, including perturbation of plant water status. These effects were assessed by exposing the unique Cd-tolerant and Cd-accumulating pea (Pisum sativum L.) mutant SGECd(t) and its wild-type (WT) line SGE to either cadmium (1, 4 μM CdCl2) or mercury (0.5, 1, 2 μM HgCl2) in hydroponic culture for 12 days. When exposed to Cd, SGECd(t) accumulated more Cd in roots, xylem sap, and shoot, and had considerably more biomass than WT plants. WT plants lost circa 0.2 MPa turgor when grown in 4 μM CdCl2, despite massive decreases in whole-plant transpiration rate and stomatal conductance. In contrast, root Hg accumulation was similar in both genotypes, but WT plants accumulated more Hg in leaves and had a higher stomatal conductance, and root and shoot biomass compared with SGECd(t). Shoot excision resulted in greater root-pressure induced xylem exudation of SGECd(t) in the absence of Cd or Hg and following Cd exposure, whereas the opposite response or no genotypic differences occurred following Hg exposure. Exposing plants that had not been treated with metal to 50 μM CdCl2 for 1h increased root xylem exudation of WT, whereas 50 μM HgCl2 inhibited and eliminated genotypic differences in root xylem exudation, suggesting differences between WT and SGECd(t) plants in aquaporin function. Thus, root water transport might be involved in mechanisms of increased tolerance and accumulation of Cd in the SGECd(t) mutant. However, the lack of cross-tolerance to Cd and Hg stress in the mutant indicates metal-specific mechanisms related to plant adaptation. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  10. Phenotypic characterization of the CRISPA (ARP gene) mutant of pea (Pisum sativum; Fabaceae): a reevaluation.

    PubMed

    DeMason, Darleen A; Chetty, Venkateswari

    2014-03-01

    Leaf form and development are controlled genetically. The ARP genes encode MYB transcription factors that interact with Class 1 KNOX genes in a regulatory module that controls meristem-leaf determinations and is highly conserved in plants. ARP loss of function alleles and subsequent KNOX1 overexpression cause many unusual leaf phenotypes including loss or partial loss of the ability to produce a lamina and production of "knots" on leaf blades. CRISPA (CRI) is the ARP gene in pea, and a number of its mutant alleles are known. We made morphological and anatomical evaluations of cri-1 mutant plants while controlling for genetic background and for heteroblastic effects, and we used aldehyde fixation and resin preparations for anatomical analysis. Further, we compared gene expression in WT and cri-1 shoot tips and HOP1/PsKN1 and CRI expression in other leaf mutants. The cri-1 plants had more extensive abnormalities in the proximal than in the distal regions of the leaf, including ectopic stipules, narrow leaflets, and shortened petioles with excessive adaxial expansion. "Knots" were morphologically and anatomically variable but consisted of vascularized out-pocketing of the adaxial leaflet surface. HOP1/PsKN1 and UNI mRNA levels were higher in cri-1 shoot tips, and some auxin-regulated genes were lower. Low LE expression suggests that the GA level is high in cri-1 shoot tips. The CRISPA gene of pea suppresses KNOX1 genes and UNI and functions to (1) maintain proximal-distal regions in their appropriate positions, (2) restrict excessive adaxial cell proliferation, and (3) promote laminar expansion.

  11. [Features of Expression of the PsSst] and PsIgn1 Genes in Nodules of Pea (Pisum sativum L.) Symbiotic Mutants].

    PubMed

    Zhukova, V A; Rychagova, T S; Fedorina, Ya V; Pinaeva, A G; Andronova, E E; Borisova, A Yu; Tikhonovich, I A

    2016-04-01

    The sequences of the PsSst1 and PsIgn1 genes of pea (Pisum sativum L.) homologous to the symbiotic LjSST1 and LjIGN1 genes of Lotusjaponicus (Regel.) K. Larsen are determined. The expression level of PsSst1 and PsIgn1 genes is determined by real-time PCR in nodules of several symbiotic mutants and original lines of pea. Lines with increased (Sprint-2Fix⁻ (Pssym31)) and decreased (P61 (Pssym25)) expression level of both genes are revealed along with the lines characterized by changes in the expression level of only one of these genes. The revealed features of the PsSst1 and PsIgn1 expression allow us to expand the phenotypic characterization of pea symbiotic mutants. In addition, PsSst1 and PsIgn1 cDNA is sequenced in selected mutant lines, characterized by a decreased expression level of these genes in nodules, but no mutations are found.

  12. Pea, Pisum sativum, and Its Anticancer Activity

    PubMed Central

    Rungruangmaitree, Runchana; Jiraungkoorskul, Wannee

    2017-01-01

    Pisum sativum (Family: Fabaceae), as known as green pea or garden pea, has long been important in diet due to its content of fiber, protein, starch, trace elements, and many phytochemical substances. It has been shown to possess antibacterial, antidiabetic, antifungal, anti-inflammatory, antihypercholesterolemia, and antioxidant activities and also shown anticancer property. Its nonnutritive biologically active components include alkaloids, flavonoids, glycosides, isoflavones, phenols, phytosterols, phytic acid, protease inhibitors, saponins, and tannins. This plant is rich in apigenin, hydroxybenzoic, hydroxycinnamic, luteolin, and quercetin, all of which have been reported to contribute to its remedial properties including anticarcinogenesis property. Based on established literature on the anticancer property of P. sativum and possible mode of action, this review article has focused to demonstrate that P. sativum could be further explored for the development of anticancer treatment. PMID:28503053

  13. Pea, Pisum sativum, and Its Anticancer Activity.

    PubMed

    Rungruangmaitree, Runchana; Jiraungkoorskul, Wannee

    2017-01-01

    Pisum sativum (Family: Fabaceae), as known as green pea or garden pea, has long been important in diet due to its content of fiber, protein, starch, trace elements, and many phytochemical substances. It has been shown to possess antibacterial, antidiabetic, antifungal, anti-inflammatory, antihypercholesterolemia, and antioxidant activities and also shown anticancer property. Its nonnutritive biologically active components include alkaloids, flavonoids, glycosides, isoflavones, phenols, phytosterols, phytic acid, protease inhibitors, saponins, and tannins. This plant is rich in apigenin, hydroxybenzoic, hydroxycinnamic, luteolin, and quercetin, all of which have been reported to contribute to its remedial properties including anticarcinogenesis property. Based on established literature on the anticancer property of P. sativum and possible mode of action, this review article has focused to demonstrate that P. sativum could be further explored for the development of anticancer treatment.

  14. E151 (sym15), a pleiotropic mutant of pea (Pisum sativum L.), displays low nodule number, enhanced mycorrhizae, delayed lateral root emergence, and high root cytokinin levels.

    PubMed

    Jones, James M C; Clairmont, Lindsey; Macdonald, Emily S; Weiner, Catherine A; Emery, R J Neil; Guinel, Frédérique C

    2015-07-01

    In legumes, the formation of rhizobial and mycorrhizal root symbioses is a highly regulated process which requires close communication between plant and microorganism. Plant mutants that have difficulties establishing symbioses are valuable tools for unravelling the mechanisms by which these symbioses are formed and regulated. Here E151, a mutant of Pisum sativum cv. Sparkle, was examined to characterize its root growth and symbiotic defects. The symbioses in terms of colonization intensity, functionality of micro-symbionts, and organ dominance were compared between the mutant and wild type. The endogenous cytokinin (CK) and abscisic acid (ABA) levels and the effect of the exogenous application of these two hormones were determined. E151 was found to be a low and delayed nodulator, exhibiting defects in both the epidermal and cortical programmes though a few mature and functional nodules develop. Mycorrhizal colonization of E151 was intensified, although the fungal functionality was impaired. Furthermore, E151 displayed an altered lateral root (LR) phenotype compared with that of the wild type whereby LR emergence is initially delayed but eventually overcome. No differences in ABA levels were found between the mutant and the wild type, but non-inoculated E151 exhibited significantly high CK levels. It is hypothesized that CK plays an essential role in differentially mediating the entry of the two micro-symbionts into the cortex; whereas it would inhibit the entry of the rhizobia in that tissue, it would promote that of the fungus. E151 is a developmental mutant which may prove to be a useful tool in further understanding the role of hormones in the regulation of beneficial root symbioses. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  15. E151 (sym15), a pleiotropic mutant of pea (Pisum sativum L.), displays low nodule number, enhanced mycorrhizae, delayed lateral root emergence, and high root cytokinin levels

    PubMed Central

    Jones, James M. C.; Clairmont, Lindsey; Macdonald, Emily S.; Weiner, Catherine A.; Emery, R. J. Neil; Guinel, Frédérique C.

    2015-01-01

    In legumes, the formation of rhizobial and mycorrhizal root symbioses is a highly regulated process which requires close communication between plant and microorganism. Plant mutants that have difficulties establishing symbioses are valuable tools for unravelling the mechanisms by which these symbioses are formed and regulated. Here E151, a mutant of Pisum sativum cv. Sparkle, was examined to characterize its root growth and symbiotic defects. The symbioses in terms of colonization intensity, functionality of micro-symbionts, and organ dominance were compared between the mutant and wild type. The endogenous cytokinin (CK) and abscisic acid (ABA) levels and the effect of the exogenous application of these two hormones were determined. E151 was found to be a low and delayed nodulator, exhibiting defects in both the epidermal and cortical programmes though a few mature and functional nodules develop. Mycorrhizal colonization of E151 was intensified, although the fungal functionality was impaired. Furthermore, E151 displayed an altered lateral root (LR) phenotype compared with that of the wild type whereby LR emergence is initially delayed but eventually overcome. No differences in ABA levels were found between the mutant and the wild type, but non-inoculated E151 exhibited significantly high CK levels. It is hypothesized that CK plays an essential role in differentially mediating the entry of the two micro-symbionts into the cortex; whereas it would inhibit the entry of the rhizobia in that tissue, it would promote that of the fungus. E151 is a developmental mutant which may prove to be a useful tool in further understanding the role of hormones in the regulation of beneficial root symbioses. PMID:25948707

  16. Characterization and structural analysis of wild type and a non-abscission mutant at the development funiculus (Def) locus in Pisum sativum L

    PubMed Central

    Ayeh, Kwadwo Owusu; Lee, YeonKyeong; Ambrose, Mike J; Hvoslef-Eide, Anne Kathrine

    2009-01-01

    Background In pea seeds (Pisum sativum L.), the Def locus defines an abscission event where the seed separates from the funicle through the intervening hilum region at maturity. A spontaneous mutation at this locus results in the seed failing to abscise from the funicle as occurs in wild type peas. In this work, structural differences between wild type peas that developed a distinct abscission zone (AZ) between the funicle and the seed coat and non-abscission def mutant were characterized. Results A clear abscission event was observed in wild type pea seeds that were associated with a distinct double palisade layers at the junction between the seed coat and funicle. Generally, mature seeds fully developed an AZ, which was not present in young wild type seeds. The AZ was formed exactly below the counter palisade layer. In contrast, the palisade layers at the junction of the seed coat and funicle were completely absent in the def mutant pea seeds and the cells in this region were seen to be extensions of surrounding parenchymatous cells. Conclusion The Def wild type developed a distinct AZ associated with palisade layer and counterpalisade layer at the junction of the seed coat and funicle while the def mutant pea seed showed non-abscission and an absence of the double palisade layers in the same region. We conclude that the presence of the double palisade layer in the hilum of the wild type pea seeds plays an important structural role in AZ formation by delimiting the specific region between the seed coat and the funicle and may play a structural role in the AZ formation and subsequent detachment of the seed from the funicle. PMID:19549315

  17. Seed coat import and unloading in pisum. [Pisum sativum

    SciTech Connect

    Grusak, M.A.; Minchin, P.E.H.

    1987-08-01

    Experiments were undertaken with empty, attached ovules of Pisum sativum to observe the effects of osmotic solution changes on seed coat import and unloading into the apoplast. Through the use of /sup 11/CO/sub 2/ pulse labelling along with collimated monitoring of plant sections, the authors were able to continuously and simultaneously measure total pod import, import into a single ovule, and washout from the ovule into a flow-through bathing solution. The authors results indicated that changes in bathing solution sucrose concentration had no immediate effect on tracer washout in Pisum, but did affect ovule import. Lowering the sucrose concentration decreased import and raising the concentration increased import. Furthermore, these import changes were only gradually reflected in the seed coat washout profile, suggesting a buffering capability of the non-phloem seed coat tissues. Additional results have also led them to propose that the terminal site of seed coat unloading in Pisum is the plasmalemma of an non-phloem seed coat cell type, that unloading from this site occurs via a passive membrane transport process, and that solutes move symplastically to this compartment from the phloem.

  18. On the shock response of pisum sativum and lepidium sativum

    NASA Astrophysics Data System (ADS)

    Leighs, James Allen; Hazell, Paul; Appleby-Thomas, Gareth James

    2012-03-01

    The high strain-rate response of biological and organic structures is of interest to numerous fields ranging from the food industry to astrobiology. Consequently, knowledge of the damage mechanisms within, and the viability of shocked organic material are of significant importance. In this study, a single-stage gasgun has been employed to subject samples of Pisum sativum (common pea) and Lepidium sativum (curled cress) to planar shock loading. Impact pressures of up to ~11.5 GPa and ~0.5 GPa for pea and cress seed samples respectively have been reached. The development of the experimental approach is discussed and presented alongside results from modelled gauge traces showing the sample loading history. Viability of the shock-loaded pea and cress seeds was investigated via attempts at germination, which were unsuccessful with pea seeds but successful in all tests performed on cress seeds. This work suggests that organic structures could survive shockwaves that may be encountered during asteroid collisions.

  19. Biosynthesis of the phytoalexin pisatin. [Pisum sativum

    SciTech Connect

    Preisig, C.L.; Bell, J.N.; Matthews, D.E.; VanEtten, H.D. ); Sun, Yuejin; Hrazdina, G. )

    1990-11-01

    NADPH-dependent reduction of 2{prime},7-dihydroxy-4{prime},5{prime}-methylenedioxyisoflavone to the isoflavanone sophorol, a proposed intermediate step in pisatin biosynthesis, was detected in extracts of Pisum sativum. This isoflavone reductase activity was inducible by treatment of pea seedlings with CuCl{sub 2}. The timing of induction coincided with that of the 6a-hydroxymaackiain 3-O-methyltransferase, which catalyzes the terminal biosynthetic step. Neither enzyme was light inducible. Further NADPH-dependent metabolism of sophorol by extracts of CuCl{sub 2}-treated seedlings was also observed; three products were radiolabeled when ({sup 3}H)sophorol was the substrate, one of which is tentatively identified as maackiain.

  20. [Pigment accumulation and functional activity of chloroplasts in common Pisum sativum L. mutants with low chlorophyll level (chlorotica)].

    PubMed

    Ladygin, V G

    2003-01-01

    Pea mutants chlorotica 2004 and 2014 with a low content of chlorophyll were studied. The mutant 2004 has light green leaves and stem, and the mutant 2014 has yellow green leaves and stem. They accumulate approximately 80 and 50% chlorophylls of the parent form of pea Torsdag cv. The content of carotene in carotenoids of the mutant 2004 was much lower, and the accumulation of lutein and violaxanthine was increased. The accumulation of all carotenoids in the mutant 2014 decreased almost proportionally to a decrease in the chlorophyll content. The rate of CO2 evolution in mutant chlorotica 2004 and 2014 was established to be lower. The quantum efficiency of photosynthesis in the mutants was 29-30% lower as compared to the control, and in hybrid plants it was 1.5-2-fold higher. It is assumed that the increase in the activity of the night-time respiration in gas exchange of chlorotica mutants and the drop of photosynthesis lead to a decrease in biomass increment. The results obtained allow us to conclude that the mutation of chlorotica 2004 and 2014 affects the genes controlling the formation and functioning of different components of the photosynthetic apparatus.

  1. Carbohydrate breakdown by chloroplasts of Pisum sativum.

    PubMed

    Stitt, M; Rees, T A

    1980-01-17

    1. The aims of this work were to discover the pathways of starch breakdown and carbohydrate metabolism in intact isolated chloroplasts from shoots of Pisum sativum. 2. 14C from starch, labelled by supplying [14C]glucose to chloroplasts, appeared, during starch breakdown, in CO2, maltose and the fraction of the acidic compounds that contained 3-phosphoglycerate and sugar phosphates. 3. When intact chloroplasts were incubated in the dark, 3-phosphoglycerate, triose phosphates and, to a lesser extent, hexose 6-phosphates accumulated in the medium at rates comparable to those of starch breakdown in leaves. This accumulation was dependent upon orthophosphate. 4. The patterns of 14CO2 production from specifically labelled [14C]glucose supplied to isolated chloroplasts were those expected of the oxidative pentose phosphate pathway with extensive recycling, and glycolysis. The respone of this pattern to lack of orthophosphate, addition of unlabelled intermediates, and 2-phosphoglycollate confirmed this view. 5. Starch breakdown in pea chloroplasts is held to be dominantly phosphorolytic with the products being metabolized via the oxidative pentose phosphate pathway and glycolysis to 3-phosphoglycerate, triose phosphates and CO2 that are exported to the cytoplasm.

  2. Protein methylation in pea chloroplasts. [Pisum sativum

    SciTech Connect

    Niemi, K.J.; Adler, J.; Selman, B.R. )

    1990-07-01

    The methylation of chloroplast proteins has been investigated by incubating intact pea (Pisum sativum) chloroplasts with ({sup 3}H-methyl)-S-adenosylmethionine. Incubation in the light increases the amount of methylation in both the thylakoid and stromal fractions. Numerous thylakoid proteins serve as substrates for the methyltransfer reactions. Three of these thylakoid proteins are methylated to a significantly greater extent in the light than in the dark. The primary stromal polypeptide methylated is the large subunit of ribulose bisphosphate carboxylase/oxygenase. One other stromal polypeptide is also methylated much more in the light than in the dark. Two distinct types of protein methylation occur. One methylinkage is stable to basic conditions whereas a second type is base labile. The base-stable linkage is indicative of N-methylation of amino acid residues while base-lability is suggestive of carboxymethylation of amino acid residues. Labeling in the light increases the percentage of methylation that is base labile in the thylakoid fraction while no difference is observed in the amount of base-labile methylations in light-labeled and dark-labeled stromal proteins. Also suggestive of carboxymethylation is the detection of volatile ({sup 3}H)methyl radioactivity which increases during the labeling period and is greater in chloroplasts labeled in the light as opposed to being labeled in the dark; this implies in vivo turnover of the ({sup 3}H)methyl group.

  3. Pea (Pisum sativum L.) in the genomics era

    USDA-ARS?s Scientific Manuscript database

    Pea (Pisum sativum L.) was the original model organism for Mendel´s discovery of the laws of inheritance, making it the foundation of modern plant genetics. However, subsequent progress in pea genomics has lagged behind many other plant species, largely as a consequence of its low multiplication rat...

  4. Gravitropic response and circumnutation in pea (Pisum sativum) seedling roots.

    PubMed

    Kim, Hye-jeong; Kobayashi, Akie; Fujii, Nobuharu; Miyazawa, Yutaka; Takahashi, Hideyuki

    2016-05-01

    Plant circumnutation is a helical movement of growing organs such as shoots and roots. Gravitropic response is hypothesized to act as an external oscillator in shoot circumnutation, although this is subject to debate. The relationship between circumnutational movement and gravitropic response in roots remains unknown. In this study, we analyzed circumnutation of agravitropic roots using the ageotropum pea (Pisum sativum) mutant, and compared it with that of wild-type (cv. Alaska) pea roots. We further examined the relationship of gravitropic response to circumnutation of Alaska seedling roots by removing the gravisensing tissue (the root cap) and by treating the roots with auxin transport inhibitors. Alaska roots displayed circumnutational movements with a period of approximately 150 min, whereas ageotropum roots did not exhibit distinct circumnutational movement. Removal of the root cap in Alaska roots reduced gravitropic response and circumnutational movements. Treatment of Alaska roots with auxin transport inhibitors, 2,3,5-triiodobenzoic acid (TIBA) and N-(1-naphthyl)phthalamic acid (NPA), dramatically reduced gravitropic response and circumnutational movements. These results suggest that a gravity-regulated auxin transport is involved in circumnutation of pea seedling roots. © 2015 Scandinavian Plant Physiology Society.

  5. Gravity-controlled asymmetrical transport of auxin regulates a gravitropic response in the early growth stage of etiolated pea (Pisum sativum) epicotyls: studies using simulated microgravity conditions on a three-dimensional clinostat and using an agravitropic mutant, ageotropum.

    PubMed

    Hoshino, Tomoki; Miyamoto, Kensuke; Ueda, Junichi

    2007-09-01

    Increased expression of the auxin-inducible gene PsIAA4/5 was observed in the elongated side of epicotyls in early growth stages of etiolated pea (Pisum sativum L. cv. Alaska) seedlings grown in a horizontal or an inclined position under 1 g conditions. Under simulated microgravity conditions on a 3D clinostat, accumulation of PsIAA4/5 mRNA was found throughout epicotyls showing automorphosis. Polar auxin transport in the proximal side of epicotyls changed when the seedlings were grown in a horizontal or an inclined position under 1 g conditions, but that under clinorotation did not, regardless of the direction of seed setting. Accumulation of PsPIN1 and PsPIN2 mRNAs in epicotyls was affected by gravistimulation, but not by clinorotation. Under 1 g conditions, auxin-transport inhibitors made epicotyls of seedlings grown in a horizontal or inclined position grow toward the proximal direction to cotyledons. These inhibitors led to epicotyl bending toward the cotyledons in seedlings grown in an inclined position under clinorotation. Polar auxin transport, as well as growth direction, of epicotyls of the agravitropic mutant ageotropum did not respond to various gravistimulation. These results suggest that alteration of polar auxin transport in the proximal side of epicotyls regulates the graviresponse of pea epicotyls.

  6. Plastid inheritance in Pisum sativum L.

    PubMed

    Polans, N O; Corriveau, J L; Coleman, A W

    1990-12-01

    Cultivar variability for levels of plastid DNA (cpDNA) in the germ cell line of germinated pea pollen has suggested the possibility of biparental plastid transmission. In order to examine this possibility further, RFLP markers were used to follow the transmission of cpDNA from parents to their F1 offspring. Results from these inheritance studies clearly indicate the presence of only maternal plastid markers in the F1 progeny of each cross examined, irrespective of the pollen cpDNA levels of the paternal parent. The same result is obtained for F1 progeny produced from crosses using pollen characterized by comparatively high cpDNA content, even when offspring are sampled at early developmental stages. Thus, there appears to be little correspondence between pollen cytological data indicating potential paternal plastid transmission and data from molecular marker studies confirming that P. sativum generally follows a uniparental-maternal mode of plastid inheritance. Insufficient F1 progeny were examined to exclude instances of trace biparentalism.

  7. Stomatal responses to carbon dioxide of isolated epidermis from a C3 plant, the Argenteum mutant of Pisum sativum L., and a crassulacean-acid-metabolism plant Kalanchoë daigremontiana Hamet et Perr.

    PubMed

    Jewer, P C; Neales, T F; Incoll, L D

    1985-07-01

    The response of stomata in isolated epidermis to the concentration of CO2 in the gaseous phase was examined in a C3 species, the Argenteum mutant of Pisum sativum, and a crassulacean-acid-metabolism (CAM) species, Kalanchoë daigremontiana. Epidermis from leaves of both species was incubated on buffer solutions in the presence of air containing various volume fractions of CO2 (0 to 10000·10(-6)). In both species and in the light and in darkness, the effect of CO2 was to inhibit stomatal opening, the maximum inhibition of opening occurring in the range 0 to 360·10(-6). The inhibition of opening per unit change in concentration was greatest between volume fractions of 0 and 240·10(-6). There was little further closure above the volume fraction of 360·10(-6), i.e. approximately ambient concentration of CO2. Thus, although leaves of CAM species may experience much higher internal concentrations of CO2 in the light than those of C3 plants, this does not affect the sensitivity of their stomata to CO2 concentration or the range over which they respond. Stomatal responses to CO2 were similar in both the light and the dark, indicating that effects of CO2 on stomata occur via mechanisms which are independent of light. The responses of stomata to CO2 in the gaseous phase took place without the treatments changing the pH of the buffered solutions. Thus it is unlikely that CO2 elicited stomatal movement by changing either the pH or the HCO 3 (-) /CO 3 (2-) equilibria. It is suggested that the concentration of dissolved unhydrated CO2 may be the effector of stomatal movement and that its activity is related to its reactivity with amines.

  8. Auxin transport inhibitor induced low complexity petiolated leaves and sessile leaf-like stipules and architectures of heritable leaf and stipule mutants in Pisum sativum suggest that its simple lobed stipules and compound leaf represent ancestral forms in angiosperms.

    PubMed

    Kumar, Arvind; Sharma, Vishakha; Khan, Moinuddin; Hindala, Mali Ram; Kumar, Sushil

    2013-04-01

    In angiosperms, leaf and stipule architectures are inherited species-specific traits. Variation in leaf and stipule sizes, and forms result from the interaction between abiotic and biotic stimuli, and gene regulatory network(s) that underlie the leaf and stipule developmental programme(s). Here, correspondence between variation in leaf and stipule architectures described for extant angiosperms and that induced mutationally and by imposition of stress in model angiosperm species, especially in Pisum sativum, was detected. Following inferences were drawn from the observations. (i) Several leaf forms in P. sativum have origin in fusion of stipule and leaf primordia. Perfoliate (and amplexicaul and connate) simple sessile leaves and sessile adnate leaves are the result of such primordial fusions. Reversal of changes in the gene regulatory network responsible for fusion products are thought to restore original stipule and leaf conditions. (ii) Compound leaf formation in several different model plants, is a result of promotion of pathways for such condition by gene regulatory networks directed by KNOx1 and LEAFY transcription factors or intercalation of the gene networks directed by them. (iii) Gene regulatory network for compound leaves in P. sativum when mutated generates highly complex compound leaves on one hand and simple leaves on other hand. These altered conditions are mutationally reversible. (vi) Simple leaves in model plants such as Arabidopsis thaliana despite overexpression of KNOx1 orthologues do not become compound. (v) All forms of leaves, including simple leaf, probably have origins in a gene regulatory network of the kind present in P. sativum.

  9. Pea weevil, Bruchus pisorum (L.) (Coleoptera: Bruchidae), resistance in Pisum sativum x P. fulvum interspecific crosses

    USDA-ARS?s Scientific Manuscript database

    The pea weevil, Bruchus pisorum (L.), is one of the most intractable pest problems of cultivated pea, Pisum sativum L., in the world. This study investigated the transfer of pea weevil resistance from two accessions (PI 595946, PI 343955) of wild pea, Pisum fulvum Sibth. & Sm., to interspecific pop...

  10. Chemical composition and pharmacological activities of Pisum sativum.

    PubMed

    Zilani, Md Nazmul Hasan; Sultana, Tamanna; Asabur Rahman, S M; Anisuzzman, Md; Islam, Md Amirul; Shilpi, Jamil A; Hossain, Md Golam

    2017-03-27

    Consumption of vegetables has been proven to be effective in the prevention of different diseases. Traditionally edible aerial part of Pisum sativum L. subsp. sativum (Fabaceae) is used to treat diabetes, heart diseases and as blood purifier. Present study was aimed to explore the traditional use of aerial parts of P. sativum as a source of antidiabetic agent. In addition, antioxidant activity and chemical composition was carried out. Total polyphenol content was spectrophotometrically determined using Folin Chiocalteu's reagent while the flavonoids by aluminum chloride colorimetric assay. Identification of compounds of the extract was made through HPLC and LCMS. Antihyperglycemic activity was assessed by oral glucose tolerance test in mice. Antioxidant activity was determined by DPPH free radical scavenging and reducing power assay. Total polyphenol and total flavonoids content were found to be 51.23 mg gallic acid equivalent and 30.88 mg quercetin equivalent per gram of dried plant extract respectively. Ellagic acid and p-coumeric acid were detected through HPLC. A total of eight compounds including naringenin, β-sitosterol were indentified through LCMS. In OGTT, extract (200 mg/kg bw) showed a 30.24% decrease (P< 0.05) in blood glucose levels at 30 min compared to the normal control. The extract showed IC50 value of 158.52 μg/mL in DPPH scavenging assay and also showed comparable reducing power. Along with other compounds ellagic acid and β-sitosterol present in the extract may be responsible for its antioxidant as well as antihyperglycemic activities. Altogether these results rationalize the use of this vegetable in traditional medicine.

  11. Enhancing phytoremediative ability of Pisum sativum by EDTA application.

    PubMed

    Piechalak, Aneta; Tomaszewska, Barbara; Barałkiewicz, Danuta

    2003-12-01

    The aim of our research was to demonstrate how the presence of EDTA affects resistance of pea plants to Pb and Pb-EDTA presence, and to show the effectivity of lead ions accumulation and translocation. It was determined that EDTA not only increased the amount of Pb taken up by plants but also Pb ion transport through the xylem and metal translocation from roots to stems and leaves. It can be seen in the presented research results that addition of the chelator with Pb limited metal phytotoxicity. We also demonstrated a significant effect of EDTA not only on Pb accumulation and metal transport to the aboveground parts but also on the profile and amount of thiol compounds: glutathione (GSH), homoglutathione (hGSH) or phytochelatins (PCs), synthesized by the plants. We observed a significant effect of the synthetic chelator on increasing the level of Pb accumulation in roots of plants treated with Pb including EDTA (0.5 and 1 mM). Pisum sativum plants treated only with 1 mM Pb(NO3)2 accumulated over 50 mg Pb x g(-1) dry wt during 4 days of cultivation. Whereas in roots of pea plants exposed to Pb+0.5 mM EDTA 35% more Pb was observed. When 1 mM EDTA was applied roots of pea accumulated over 67% more metal. The presence of EDTA also increased metal uptake and transport to the aboveground parts. In pea plants treated only with 1 mM lead nitrate less than 3 mg Pb x g(-1) dry wt was transported, whereas in P. sativum treated with Pb-EDTA doubled amount of Pb was observed in stems and leaves.

  12. Identification of the 64 kilodalton chloroplast stromal phosphoprotein as phosphoglucomutase. [Pisum sativum

    SciTech Connect

    Salvucci, M.E.; Drake, R.R.; Broadbent, K.P.; Haley, B.E. ); Hanson, K.R.; McHale, N.A. )

    1990-05-01

    Phosphorylation of the 64 kilodalton stromal phosphoprotein by incubation of pea (Pisum sativum) chloroplast extracts with ({gamma}-{sup 32}P)ATP decreased in the presence of Glc-6-P and Glc-1,6-P{sub 2}, but was stimulated by glucose. Two-dimensional gel electrophoresis following incubation of intact chloroplasts and stromal extracts with ({gamma}-{sup 32}P)ATP, or incubation of stromal extracts and partially purified phosphoglucomutase (EC 2.7.5.1) with ({sup 32}P)Glc-1-P showed that the identical 64 kilodalton polypeptide was labeled. A 62 kilodalton polypeptide was phosphorylated by incubation of tobacco (Nicotiana sylvestris) stromal extracts with either ({gamma}-{sup 32}P)ATP or ({sup 32}P)Glc-1-P. In contrast, an analogous polypeptide was not phosphorylated in extracts from a tobacco mutant deficient in plastid phosphoglucomutase activity. The results indicate that the 64 (or 62) kilodalton chloroplast stromal phosphoprotein is phosphoglucomutase.

  13. Interdependence of Nitrogen Nutrition and Photosynthesis in Pisum sativum L

    PubMed Central

    Bethlenfalvay, Gabor J.; Abu-Shakra, Salah S.; Phillips, Donald A.

    1978-01-01

    Physiological responses to infection by strains of Rhizobium leguminosarum which differed in their capacity to reduce N2 were determined in 26-day-old pea plants (Pisum sativum L. cv. Alaska) grown under uniform environmental conditions in the absence of combined N. The highest N2 reduction rates, calculated from H2 evolution and C2H2-dependent C2H4 production measurements, were approximately 6-fold greater than the lowest. Higher N2 fixation rates were associated with greater CO2 exchange rates (R2 = 0.92) and carboxylation efficiency (R2 = 0.99). Increases in the apparent relative efficiency of N2 fixation [1-(H2 evolved in air/C2H2 reduced)] (acteroid efficiency) were associated with increases in whole-plant N2 fixation efficiency (N2/CO2 reduction ratio) (R2 = 0.95). Whole-plant dry weight and total N content were related by regression analysis (R2 = 0.98); both parameters were enhanced by increased N2 fixation in a manner analogous to previously reported increases caused by greater external applications of NH4+. These data reveal that photosynthetic parameters in genetically uniform host plants grown under identical environmental conditions are affected by N2 fixation characteristics of the rhizobial symbiont. The measured efficiencies of micro- and macrosymbiont are directly related under uniform environments. PMID:16660451

  14. Further characterization of ribosome binding to thylakoid membranes. [Pisum sativum

    SciTech Connect

    Hurewitz, J.; Jagendorf, A.T.

    1987-05-01

    Previous work indicated more polysomes bound to pea (Pisum sativum cv Progress No. 9) thylakoids in light than in the dark, in vivo. With isolated intact chloroplasts incubated in darkness, addition of MgATP had no effect but 24 to 74% more RNA was thylakoid-bound at pH 8.3 than at pH 7. Thus, the major effect of light on ribosome-binding in vivo may be due to higher stroma pH. In isolated pea chloroplasts, initiation inhibitors (pactamycin and kanamycin) decreased the extent of RNA binding, and elongation inhibitors (lincomycin and streptomycin) increased it. Thus, cycling of ribosomes is controlled by translation, initiation, and termination. Bound RNA accounted for 19 to 24% of the total chloroplast RNA and the incorporation of (/sup 3/H)leucine into thylakoids was proportional to the amount of this bound RNA. These data support the concept that stroma ribosomes are recruited into thylakoid polysomes, which are active in synthesizing thylakoid proteins.

  15. Burdock fructooligosaccharide induces stomatal closure in Pisum sativum.

    PubMed

    Guo, Yanling; Guo, Moran; Zhao, Wenlu; Chen, Kaoshan; Zhang, Pengying

    2013-09-12

    Burdock fructooligosaccharide (BFO) isolated from the root tissue of Arctium lappa is a reserve carbohydrate that can induce resistance against a number of plant diseases. Stomatal closure is a part of plant innate immune response to restrict bacterial invasion. In this study, the effects of BFO on stomata movement in Pisum sativum and the possible mechanisms were studied with abscisic acid (ABA) as a positive control. The results showed that BFO could induce stomatal closure accompanied by ROS and NO production, as is the case with ABA. BFO-induced stomatal closure was inhibited by pre-treatment with L-NAME (N(G)-nitro-L-arginine methyl ester, hydrochloride; nitric oxide synthase inhibitor) and catalase (hydrogen peroxide scavenger). Exogenous catalase completely restricted BFO-induced production of ROS and NO in guard cells. In contrast, L-NAME prevented the rise in NO levels but only partially restricted the ROS production. These results indicate that BFO-induced stomatal closure is mediated by ROS and ROS-dependent NO production.

  16. Characterization of pea (Pisum sativum) seed protein fractions.

    PubMed

    Rubio, Luis A; Pérez, Alicia; Ruiz, Raquel; Guzmán, M Ángeles; Aranda-Olmedo, Isabel; Clemente, Alfonso

    2014-01-30

    Legume seed proteins have to be chemically characterized in order to properly link their nutritional effects with their chemical structure. Vicilin and albumin fractions devoid of cross-contamination, as assessed by mass peptide fingerprinting analysis, were obtained from defatted pea (Pisum sativum cv. Bilbo) meal. The extracted protein fractions contained 56.7-67.7 g non-starch polysaccharides kg⁻¹. The vicilin fraction was higher than legumins in arginine, isoleucine, leucine, phenylalanine and lysine. The most abundant amino acids in the albumin fraction were aspartic acid, glutamic acid, lysine and arginine, and the amounts of methionine were more than double than those in legumins and vicilins. The pea albumin fraction showed a clear enrichment of protease inhibitory activity when compared with the seed meal. In vitro digestibility values for pea proteins were 0.63 ±  0.04, 0.88 ±  0.04 and 0.41 ±  0.23 for legumins, vicilins and albumins respectively. Vicilin and albumin fractions devoid of cross-contamination with other proteins were obtained from pea seed meal. The vicilin fraction also contained low amounts of soluble non-starch polysaccharides and was enriched in isoleucine, leucine, phenylalanine and lysine. In vitro digestibility values for pea proteins were similar or even numerically higher than those for control proteins. © 2013 Society of Chemical Industry.

  17. A polygalacturonase localized in the Golgi apparatus in Pisum sativum.

    PubMed

    Ohashi, Takao; Jinno, Jun; Inoue, Yoshiyuki; Ito, Shoko; Fujiyama, Kazuhito; Ishimizu, Takeshi

    2017-02-23

    Pectin is a plant cell wall constituent that is mainly composed of polygalacturonic acid (PGA), a linear α1,4-D-galacturonic acid (GalUA) backbone. Polygalacturonase (PG) hydrolyzes the α1,4-linkages in PGA. Nearly all plant PGs identified thus far are secreted as soluble proteins. Here we describe the microsomal PG activity in pea (Pisum sativum) epicotyls and present biochemical evidence that it was localized to the Golgi apparatus, where pectins are biosynthesized. The microsomal PG was purified, and it was enzymatically characterized. The purified enzyme showed maximum activity towards pyridylaminated oligogalacturonic acids with six degrees of polymerization (PA-GalUA6), with a Km value of 11 μM for PA-GalUA6. The substrate preference of the enzyme was complementary to that of PGA synthase. The main PG activity in microsomes was detected in the Golgi fraction by sucrose density gradient ultracentrifugation. The activity of the microsomal PG was lower in rapidly growing epicotyls, in contrast to the high expression of PGA synthase. The role of this PG in the regulation of pectin biosynthesis or plant growth is discussed. © The Authors 2017. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  18. Review of the health benefits of peas (Pisum sativum L.).

    PubMed

    Dahl, Wendy J; Foster, Lauren M; Tyler, Robert T

    2012-08-01

    Pulses, including peas, have long been important components of the human diet due to their content of starch, protein and other nutrients. More recently, the health benefits other than nutrition associated with pulse consumption have attracted much interest. The focus of the present review paper is the demonstrated and potential health benefits associated with the consumption of peas, Pisum sativum L., specifically green and yellow cotyledon dry peas, also known as smooth peas or field peas. These health benefits derive mainly from the concentration and properties of starch, protein, fibre, vitamins, minerals and phytochemicals in peas. Fibre from the seed coat and the cell walls of the cotyledon contributes to gastrointestinal function and health, and reduces the digestibility of starch in peas. The intermediate amylose content of pea starch also contributes to its lower glycaemic index and reduced starch digestibility. Pea protein, when hydrolysed, may yield peptides with bioactivities, including angiotensin I-converting enzyme inhibitor activity and antioxidant activity. The vitamin and mineral contents of peas may play important roles in the prevention of deficiency-related diseases, specifically those related to deficiencies of Se or folate. Peas contain a variety of phytochemicals once thought of only as antinutritive factors. These include polyphenolics, in coloured seed coat types in particular, which may have antioxidant and anticarcinogenic activity, saponins which may exhibit hypocholesterolaemic and anticarcinogenic activity, and galactose oligosaccharides which may exert beneficial prebiotic effects in the large intestine.

  19. Transient protein expression in three Pisum sativum (green pea) varieties.

    PubMed

    Green, Brian J; Fujiki, Masaaki; Mett, Valentina; Kaczmarczyk, Jon; Shamloul, Moneim; Musiychuk, Konstantin; Underkoffler, Susan; Yusibov, Vidadi; Mett, Vadim

    2009-02-01

    The expression of proteins in plants both transiently and via permanently transformed lines has been demonstrated by a number of groups. Transient plant expression systems, due to high expression levels and speed of production, show greater promise for the manufacturing of biopharmaceuticals when compared to permanent transformants. Expression vectors based on a tobacco mosaic virus (TMV) are the most commonly utilized and the primary plant used, Nicotiana benthamiana, has demonstrated the ability to express a wide range of proteins at levels amenable to purification. N. benthamiana has two limitations for its use; one is its relatively slow growth, and the other is its low biomass. To address these limitations we screened a number of legumes for transient protein expression. Using the alfalfa mosaic virus (AMV) and the cucumber mosaic virus (CMV) vectors, delivered via Agrobacterium, we were able to identify three Pisum sativum varieties that demonstrated protein expression transiently. Expression levels of 420 +/- 26.24 mg GFP/kgFW in the green pea variety speckled pea were achieved. We were also able to express three therapeutic proteins indicating promise for this system in the production of biopharmaceuticals.

  20. Valorization of the peel of pea: Pisum sativum by evaluation of its antioxidant and antimicrobial activities.

    PubMed

    Hadrich, Fatma; Arbi, Mahdi El; Boukhris, Maher; Sayadi, Sami; Cherif, Slim

    2014-01-01

    This study deals with evaluating antioxidant and antimicrobial activities of the peel of pea (Pisum sativum), with particular attention to the content of some bioactive compounds. Total content of polyphenols and flavonoids of Pisum sativum peel extracts, including a crude aqueous extract, a methanolic extract and an ethyl acetate extract was carried out according to the standard methods to assess their corresponding antioxidant activities. The organic solvents extracts antioxidant activities, determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, ferric reducing (FRAP) assay and 2,2 azinobis 3-ethylbenzo-thiozoline-6-sulfonic acid (ABTS) assay, were relatively high. The highest activity was found in ethyl acetate extract. The antimicrobial activities of extracts were also assessed. The highest MIC value was occurred with E.Coli (850 µg/ml) when using ethyl acetate extract. From the results obtained, Pisum sativum peel can be considered as a very good source of health promoting compounds.

  1. Photosynthetic Pod Wall of Pea (Pisum sativum L.)

    PubMed Central

    Atkins, Craig A.; Kuo, John; Pate, John S.; Flinn, Alastair M.; Steele, Trevor W.

    1977-01-01

    The pod wall of pea (Pisum sativum L.) was shown to contain two distinct photosynthetic layers. The outer, comprising chlorenchyma of the mesocarp, captured CO2 from the outside atmosphere; the inner, a chloroplast-containing epidermis lining the pod gas cavity, was involved in photoassimilation of the CO2 released from respiring seeds. Structural features of the pod included the thick cuticle and stomata of the outer epidermis, the inward projecting veinlets of the vascular network in the mesocarp, the sparsity of air spaces, the fiber and parenchyma layers of the endocarp, and the abundant chloroplasts, thin cuticle, and rounded outer contours of cells of the inner epidermis. The inner epidermis showed high specific activities of ribulose 1,5-diphosphate (RuDP) carboxylase (EC 4.1.1.39) and phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31), contained up to 20% of the pod's chlorophyll, and was capable of fixing 66% of the CO2 released during the photoperiod to the pod gas space by the seeds of a fully grown fruit. The in vitro carboxylation capacity of the pod exceeded the estimated gross photosynthesis of the fruit for all but the last few days of development. Chlorophyll content and carboxylation activity declined more markedly in the outer photosynthetic layers than in the inner epidermis. The ratio of activities of RuDP carboxylase to PEP carboxylase in pod extracts varied from 2.4:1 to 12:1 as against 48:1 to 156:1 in extracts of leaves. Structural and physiological properties of the pod were related to its capacity to conserve respired CO2 and provide photosynthate to developing seeds. Images PMID:16660184

  2. Flowering time adaption in Swedish landrace pea (Pisum sativum L.).

    PubMed

    Vanhala, Tytti; Normann, Kjersti R; Lundström, Maria; Weller, James L; Leino, Matti W; Hagenblad, Jenny

    2016-08-12

    Cultivated crops have repeatedly faced new climatic conditions while spreading from their site of origin. In Sweden, at the northernmost fringe of Europe, extreme conditions with temperature-limited growth seasons and long days require specific adaptation. Pea (Pisum sativum L.) has been cultivated in Sweden for millennia, allowing for adaptation to the local environmental conditions to develop. To study such adaptation, 15 Swedish pea landraces were chosen alongside nine European landraces, seven cultivars and three wild accessions. Number of days to flowering (DTF) and other traits were measured and the diversity of the flowering time genes HIGH RESPONSE TO PHOTOPERIOD (HR), LATE FLOWERING (LF) and STERILE NODES (SN) was assessed. Furthermore, the expression profiles of LF and SN were obtained. DTF was positively correlated with the length of growing season at the site of origin (GSO) of the Swedish landraces. Alleles at the HR locus were significantly associated with DTF with an average difference of 15.43 days between the two detected haplotypes. LF expression was found to have a significant effect on DTF when analysed on its own, but not when HR haplotype was added to the model. HR haplotype and GSO together explained the most of the detected variation in DTF (49.6 %). We show local adaptation of DTF, primarily in the northernmost accessions, and links between genetic diversity and diversity in DTF. The links between GSO and genetic diversity of the genes are less clear-cut and flowering time adaptation seems to have a complex genetic background.

  3. Isolation and characterization of novel EST-derived genic markers in Pisum sativum (Fabaceae).

    PubMed

    Jain, Shalu; McPhee, Kevin E

    2013-11-01

    Novel markers were developed for pea (Pisum sativum) from pea expressed sequence tags (ESTs) having significant homology to Medicago truncatula gene sequences to investigate genetic diversity, linkage mapping, and cross-species transferability. • Seventy-seven EST-derived genic markers were developed through comparative mapping between M. truncatula and P. sativum in which 75 markers produced PCR products and 33 were polymorphic among 16 pea genotypes. • The novel markers described here will be useful for future genetic studies of P. sativum; their amplification in lentil (Lens culinaris) demonstrates their potential for use in closely related species.

  4. Isolation and characterization of novel EST-derived genic markers in Pisum sativum (Fabaceae)1

    PubMed Central

    Jain, Shalu; McPhee, Kevin E.

    2013-01-01

    • Premise of the study: Novel markers were developed for pea (Pisum sativum) from pea expressed sequence tags (ESTs) having significant homology to Medicago truncatula gene sequences to investigate genetic diversity, linkage mapping, and cross-species transferability. • Methods and Results: Seventy-seven EST-derived genic markers were developed through comparative mapping between M. truncatula and P. sativum in which 75 markers produced PCR products and 33 were polymorphic among 16 pea genotypes. • Conclusions: The novel markers described here will be useful for future genetic studies of P. sativum; their amplification in lentil (Lens culinaris) demonstrates their potential for use in closely related species. PMID:25202494

  5. Pea (Pisum sativum) Seed Production as an Assay for Reproductive Effects Due to Herbicides.

    EPA Science Inventory

    Even though herbicide drift can affect plant reproduction, current plant testing protocols emphasize effects on vegetative growth. In this study, we determined whether a short–growing season plant can indicate potential effects of herbicides on seed production. Pea (Pisum sativum...

  6. Pea (Pisum sativum) Seed Production as an Assay for Reproductive Effects Due to Herbicides.

    EPA Science Inventory

    Even though herbicide drift can affect plant reproduction, current plant testing protocols emphasize effects on vegetative growth. In this study, we determined whether a short–growing season plant can indicate potential effects of herbicides on seed production. Pea (Pisum sativum...

  7. Genetic Diversity within the USDA Pisum sativum Collection for Seed Sugar Composition and Concentration

    USDA-ARS?s Scientific Manuscript database

    The USDA Pisum sativum refined core collection was evaluated for concentrationa dn composition of low molceular weight carbohydrates. Six plants each from the 120 wrinkle-seeded accessions were grown ina commercial soilless mix in a greenhouse under 16hr/8hr day/night 20/15 C°. Pods were harvested...

  8. Genetic Diversity of Chinese and Global Pea (Pisum sativum L.) Collections.

    USDA-ARS?s Scientific Manuscript database

    Pea (Pisum sativum L.) is an important food and feed legume grown across many temperate regions of the world, especially from Asia to Europe and North America. The goal of this study was to use 30 informative pea microsatellite markers to compare genetic diversity in a global core from the USDA and ...

  9. Characterization of proanthocyanidin metabolism in pea (Pisum sativum) seeds.

    PubMed

    Ferraro, Kiva; Jin, Alena L; Nguyen, Trinh-Don; Reinecke, Dennis M; Ozga, Jocelyn A; Ro, Dae-Kyun

    2014-09-16

    Proanthocyanidins (PAs) accumulate in the seeds, fruits and leaves of various plant species including the seed coats of pea (Pisum sativum), an important food crop. PAs have been implicated in human health, but molecular and biochemical characterization of pea PA biosynthesis has not been established to date, and detailed pea PA chemical composition has not been extensively studied. PAs were localized to the ground parenchyma and epidermal cells of pea seed coats. Chemical analyses of PAs from seeds of three pea cultivars demonstrated cultivar variation in PA composition. 'Courier' and 'Solido' PAs were primarily prodelphinidin-types, whereas the PAs from 'LAN3017' were mainly the procyanidin-type. The mean degree of polymerization of 'LAN3017' PAs was also higher than those from 'Courier' and 'Solido'. Next-generation sequencing of 'Courier' seed coat cDNA produced a seed coat-specific transcriptome. Three cDNAs encoding anthocyanidin reductase (PsANR), leucoanthocyanidin reductase (PsLAR), and dihydroflavonol reductase (PsDFR) were isolated. PsANR and PsLAR transcripts were most abundant earlier in seed coat development. This was followed by maximum PA accumulation in the seed coat. Recombinant PsANR enzyme efficiently synthesized all three cis-flavan-3-ols (gallocatechin, catechin, and afzalechin) with satisfactory kinetic properties. The synthesis rate of trans-flavan-3-ol by co-incubation of PsLAR and PsDFR was comparable to cis-flavan-3-ol synthesis rate by PsANR. Despite the competent PsLAR activity in vitro, expression of PsLAR driven by the Arabidopsis ANR promoter in wild-type and anr knock-out Arabidopsis backgrounds did not result in PA synthesis. Significant variation in seed coat PA composition was found within the pea cultivars, making pea an ideal system to explore PA biosynthesis. PsANR and PsLAR transcript profiles, PA localization, and PA accumulation patterns suggest that a pool of PA subunits are produced in specific seed coat cells early in

  10. Kinetic features of gravicurvature of pea (Pisum sativum) and cress (Lepidium sativum) roots

    NASA Astrophysics Data System (ADS)

    Polishchuk, O. V.

    The upper sides of roots oriented horizontally grow more rapidly than the lower sides, causing the root ultimately to grow downward; this phenomenon is known as positive gravitropism. This ability is based on implicit mechanism which is being extensively investigated. Elaborate analysis of kinetic features of gravicurvature may complement the investigation. Pea and cress roots have positive gravitropism as roots of majority of higher plants. Mainly we investigated dependence of gravicurvature angle on time of gravistimulation. Two-day-old seedlings of cress (Lepidium sativum L. cv. P896) and four-day-old pea ones (Pisum sativum L. cv. Damir-2) were placed on 1% agar medium in Petri dishes and turned on angle of gravistimulation. Then they were photographed at the same position each hour of gravistimulation. Photographs were analyzed with the help of Image Tool software program. Both pea and cress roots showed two phases of gravitropic response during gravistimulation for 6 hours when the initial angle of gravistimulation was 135 degrees. Two peaks of the rate of bending were observed. In cress roots, the first peak was much lower and the distance between the two peaks was greater than in pea roots. Curves of gravitropic bending of cress roots grown in agar had one or two inflections while in the case of roots grown on filter paper curves had no inflections. These data are in agreement with the effect of the external medium on the gravitropic curvature of rice roots reported by Staves et al. (1997). Our results may reflect the fact that at least two systems that contribute to gravicurvature may exist in roots. These systems may be ligand-receptor complexes that may be formed with different kinetics in two different regions of the root. The most probable ligand is auxin and the regions appear to be central elongation zone (CEZ) and distal elongation zone (DEZ), that were reported to be centers of tropic bending in roots. Thus, dependence of rate of root bending on

  11. Growth stimulation of dwarf peas (Pisum sativum L.) through homeopathic potencies of plant growth substances.

    PubMed

    Baumgartner, S; Thurneysen, A; Heusser, P

    2004-10-01

    Efficacy of higher homeopathic potencies is controversial. Universally accepted specific detection assays for homeopathic dilutions do not exist. Basic research has to develop a spectrum of standardized tools to investigate the mode of action and nature of homeopathic potencies. Can the shoot growth reaction of dwarf peas (gibberellin- deficient mutants) be regarded as evidence of treatment with homeopathic potencies of plant growth substances? Pea seed (Pisum sativum L. cv. Fruher Zwerg) is immersed for 24 hours in homeopathic potency or control solutions for soaking. Plants germinate and grow in a standard cultivation substrate under controlled environmental conditions. Shoot length is measured 14 days after planting. A screening of homeopathic potencies (12x-30x) of four different plant growth substances revealed biological activity of certain potency levels of gibberellin and kinetin (p < 0.05). Growth stimulation through gibberellin 17x (5 x 10(-18 M)) was assessed in six independent replications; results confirmed those of the screening (p < 0.05). The effect of gibberellin 17x seemed to weaken during the course of the experiments. The results back the hypothesis that homeopathic potencies of plant growth substances affect pea shoot growth. Dwarf peas might thus be an interesting system model for studying the action of homeopathic potencies. Further work is required to identify all boundary conditions modulating the reactivity of this system.

  12. Immunocytochemical localization of Pisum sativum TRXs f and m in non-photosynthetic tissues.

    PubMed

    Traverso, José A; Vignols, Florence; Cazalis, Roland; Serrato, Antonio J; Pulido, Pablo; Sahrawy, Mariam; Meyer, Yves; Cejudo, Francisco Javier; Chueca, Ana

    2008-01-01

    Plants are the organisms containing the most complex multigenic family for thioredoxins (TRX). Several types of TRXs are targeted to chloroplasts, which have been classified into four subgroups: m, f, x, and y. Among them, TRXs f and m were the first plastidial TRXs characterized, and their function as redox modulators of enzymes involved in carbon assimilation in the chloroplast has been well-established. Both TRXs, f and m, were named according to their ability to reduce plastidial fructose-1,6-bisphosphatase (FBPase) and malate dehydrogenase (MDH), respectively. Evidence is presented here based on the immunocytochemistry of the localization of f and m-type TRXs from Pisum sativum in non-photosynthetic tissues. Both TRXs showed a different spatial pattern. Whilst PsTRXm was localized to vascular tissues of all the organs analysed (leaves, stems, and roots), PsTRXf was localized to more specific cells next to xylem vessels and vascular cambium. Heterologous complementation analysis of the yeast mutant EMY63, deficient in both yeast TRXs, by the pea plastidial TRXs suggests that PsTRXm, but not PsTRXf, is involved in the mechanism of reactive oxygen species (ROS) detoxification. In agreement with this function, the PsTRXm gene was induced in roots of pea plants in response to hydrogen peroxide.

  13. Roots of Pisum sativum L. exhibit hydrotropism in response to a water potential gradient in vermiculite.

    PubMed

    Tsuda, Shogo; Miyamoto, Naoko; Takahashi, Hideyuki; Ishihara, Kuni; Hirasawa, Tadashi

    2003-12-01

    In the present study, root hydrotropism in an agravitropic mutant of Pisum sativum L. grown in vermiculite with a steep water potential gradient was examined. When wet and dry vermiculite were placed side by side, water diffused from the wet (-0.04 MPa) to the dry (-1.2 MPa) and a steep water potential gradient became apparent in the dry vermiculite close to the boundary between the two. The extent and location of the gradient remained stable between the fourth and sixth day after filling a box with vermiculite, and the steepest gradient (approx. 0.02 MPa mm-1) was found in the initially dry vermiculite between 60 and 80 mm from the boundary. When seedlings with 25-35 mm long roots were planted in the initially dry vermiculite near where the gradient had been established, each of the main roots elongated toward the wet vermiculite, i.e. toward the high water potential. Control roots elongated without curvature in both the wet and the dry vermiculite, in which no water potential gradient was detectable. These results show that pea roots respond to the water potential gradient around them and elongate towards the higher water potential. Therefore, positive hydrotropism occurs in vermiculite just as it does in air. Hydrotropism in soil may be significant when a steep water potential gradient is apparent, such as when drip irrigation is applied.

  14. Analysis of the state of posttranslational calmodulin methylation in developing pea plants. [Pisum sativum

    SciTech Connect

    Oh, Sukheung; Roberts, D.M. )

    1990-07-01

    A specific calmodulin-N-methyltransferase was used in a radiometric assay to analyze the degree of methylation of lysine-115 in pea (Pisum sativum) plants. Calmodulin was isolated from dissected segments of developing roots of young etiolated and green pea plants and was tested for its ability to be methylated by incubation with the calmodulin methyltransferase in the presence of ({sup 3}H)methyl-S-adenosylmethionine. By this approach, the presence of unmethylated calmodulins were demonstrated in pea tissues, and the levels of methylation varied depending on the developmental state of the tissue tested. Calmodulin methylation levels were lower in apical root segments of both etiolated and green plants, and in the young lateral roots compared with the mature, differentiated root tissues. The incorporation of methyl groups into these calmodulin samples appears to be specific for position 115 since site-directed mutants of calmodulin with substitutions at this position competitively inhibited methyl group incorporation. The present findings, combined with previous data showing differences in the ability of methylated and unmethylated calmodulins to activate pea NAD kinase raise the possibility that posttranslational methylation of calmodulin could be another mechanism for regulating calmodulin activity.

  15. Trehalose 6-phosphate is involved in triggering axillary bud outgrowth in garden pea (Pisum sativum L.).

    PubMed

    Fichtner, Franziska; Barbier, Francois F; Feil, Regina; Watanabe, Mutsumi; Annunziata, Maria Grazia; Chabikwa, Tinashe G; Höfgen, Rainer; Stitt, Mark; Beveridge, Christine A; Lunn, John E

    2017-09-04

    Trehalose 6-phosphate (Tre6P) is a signal of sucrose availability in plants, and has been implicated in regulation of shoot branching by the abnormal branching phenotypes of Arabidopsis (Arabidopsis thaliana) and maize (Zea mays) mutants with altered Tre6P metabolism. Decapitation of garden pea (Pisum sativum) plants has been proposed to release the dormancy of axillary buds lower down the stem due to changes in sucrose supply, and we hypothesized that this response is mediated by Tre6P. Decapitation led to a rapid and sustained rise in Tre6P levels in axillary buds, coinciding with the onset of bud outgrowth. This response was suppressed by simultaneous defoliation that restricts the supply of sucrose to axillary buds in decapitated plants. Decapitation also led to a rise in amino acid levels in buds, but a fall in phosphoenolpyruvate and 2-oxoglutarate. Supplying sucrose to stem node explants in vitro triggered a concentration-dependent increase in the Tre6P content of the buds that was highly correlated with their rate of outgrowth. These data show that changes in bud Tre6P levels are correlated with initiation of bud outgrowth following decapitation, suggesting that Tre6P is involved in the release of bud dormancy by sucrose. Tre6P might also be linked to a reconfiguration of carbon and nitrogen metabolism to support the subsequent growth of the bud into a new shoot. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  16. Regeneration of Pea (’Pisum sativum L’.) Plants from Shoot Apical Meristems,

    DTIC Science & Technology

    A procedure has been developed to obtain complete plants from meristems of three cultivars of Pisum sativum L. Benzyladenine (BA) alone or in...combination with naphthaleneacetic acid (NAA) at molar concentrations of 5 x .0000005 and .000001 respectively, induced shoot differentiation in meristems ...complete plant formation. Root formation, on the shoots produced by culturing meristems was induced by reculturing the shoots, 2 cm long, on half strength B5 medium supplemented with NAA at a concentration of .000001 M.

  17. Subcellular Localization of Asparaginase and Asparagine Aminotransferase in Pisum sativum Leaves 1

    PubMed Central

    Ireland, Robert J.; Joy, Kenneth W.

    1983-01-01

    Protoplasts isolated from young and mature pea leaves (Pisum sativum L.) were broken and their contents fractionated by differential centrifugation or on sucrose-density gradients. Asparaginase was found only in the cytosol of young leaves. Asparagine aminotransferase was found in both young and mature leaves and was localized exclusively in the peroxisome. This corroborates the observation that asparagine transamination is catalyzed by the serine:glyoxylate aminotransferase. PMID:16663132

  18. Changes in Leaf Proteins of Peas, Pisum sativum L., during Development on Deflorated Plants

    PubMed Central

    Malik, N. S. A.; Berrie, Alex M. M.

    1977-01-01

    The soluble (sap) proteins of leaves of pea, Pisum sativum L. cvs. Alaska and Greenfeast, allowed to develop normally or deflowered, to prevent senescence, were separated by isoelectric focusing. There was a decline in certain proteins, with increases in others as the leaves aged but preventing senescence of the whole plant did not alter the pattern of change in leaf proteins. We concluded that whole plant senescence proceeds independently of leaf senescence. PMID:16659844

  19. Arabidopsis thaliana and Pisum sativum models demonstrate that root colonization is an intrinsic trait of Burkholderia cepacia complex bacteria.

    PubMed

    Vidal-Quist, J Cristian; O'Sullivan, Louise A; Desert, Annaëlle; Fivian-Hughes, Amanda S; Millet, Coralie; Jones, T Hefin; Weightman, Andrew J; Rogers, Hilary J; Berry, Colin; Mahenthiralingam, Eshwar

    2014-02-01

    Burkholderia cepacia complex (Bcc) bacteria possess biotechnologically useful properties that contrast with their opportunistic pathogenicity. The rhizosphere fitness of Bcc bacteria is central to their biocontrol and bioremediation activities. However, it is not known whether this differs between species or between environmental and clinical strains. We investigated the ability of 26 Bcc strains representing nine different species to colonize the roots of Arabidopsis thaliana and Pisum sativum (pea). Viable counts, scanning electron microscopy and bioluminescence imaging were used to assess root colonization, with Bcc bacteria achieving mean (±sem) levels of 2.49±0.23×10(6) and 5.16±1.87×10(6) c.f.u. per centimetre of root on the A. thaliana and P. sativum models, respectively. The A. thaliana rhizocompetence model was able to reveal loss of colonization phenotypes in Burkholderia vietnamiensis G4 transposon mutants that had only previously been observed in competition experiments on the P. sativum model. Different Bcc species colonized each plant model at different rates, and no statistical difference in root colonization was observed between isolates of clinical or environmental origin. Loss of the virulence-associated third chromosomal replicon (>1 Mb DNA) did not alter Bcc root colonization on A. thaliana. In summary, Bcc bacteria possess intrinsic root colonization abilities irrespective of their species or source. As Bcc rhizocompetence does not require their third chromosomal replicon, the possibility of using synthetic biology approaches to engineer virulence-attenuated biotechnological strains is tractable.

  20. The tropic response of plant roots to oxygen: oxytropism in Pisum sativum L.

    PubMed

    Porterfield, D M; Musgrave, M E

    1998-09-01

    Plant roots are known to orient growth through the soil by gravitropism, hydrotropism, and thigmotropism. Recent observations of plant roots that developed in a microgravity environment in space suggested that plant roots may also orient their growth toward oxygen (oxytropism). Using garden pea (Pisum sativum L. cv. Weibul's Apollo) and an agravitropic mutant (cv. Ageotropum), root oxytropism was studied in the controlled environment of a microrhizotron. A series of channels in the microrhizotron allowed establishment of an oxygen gradient of 0.8 mmol mol-1 mm-1. Curvature of seedling roots was determined prior to freezing the roots for subsequent spectrophotometric determinations of alcohol dehydrogenase activity. Oxytropic curvature was observed all along the gradient in both cultivars of pea. The normal gravitropic cultivar showed a maximal curvature of 45 degrees after 48 h, while the agravitropic mutant curved to 90 degrees. In each cultivar, the amount of curvature declined as the oxygen concentration decreased, and was linearly related to the root elongation rate. Since oxytropic curvature occurred in roots exposed to oxygen concentrations that were not low enough to induce the hypoxically responsive protein alcohol dehydrogenase, we suspect that the oxygen sensor associated with oxytropism does not control the induction of hypoxic metabolism. Our results indicate that oxygen can play a critical role in determining root orientation as well as impacting root metabolic status. Oxytropism allows roots to avoid oxygen-deprived soil strata and may also be the basis of an auto-avoidance mechanism, decreasing the competition between roots for water and nutrients as well as oxygen.

  1. The tropic response of plant roots to oxygen: oxytropism in Pisum sativum L

    NASA Technical Reports Server (NTRS)

    Porterfield, D. M.; Musgrave, M. E.

    1998-01-01

    Plant roots are known to orient growth through the soil by gravitropism, hydrotropism, and thigmotropism. Recent observations of plant roots that developed in a microgravity environment in space suggested that plant roots may also orient their growth toward oxygen (oxytropism). Using garden pea (Pisum sativum L. cv. Weibul's Apollo) and an agravitropic mutant (cv. Ageotropum), root oxytropism was studied in the controlled environment of a microrhizotron. A series of channels in the microrhizotron allowed establishment of an oxygen gradient of 0.8 mmol mol-1 mm-1. Curvature of seedling roots was determined prior to freezing the roots for subsequent spectrophotometric determinations of alcohol dehydrogenase activity. Oxytropic curvature was observed all along the gradient in both cultivars of pea. The normal gravitropic cultivar showed a maximal curvature of 45 degrees after 48 h, while the agravitropic mutant curved to 90 degrees. In each cultivar, the amount of curvature declined as the oxygen concentration decreased, and was linearly related to the root elongation rate. Since oxytropic curvature occurred in roots exposed to oxygen concentrations that were not low enough to induce the hypoxically responsive protein alcohol dehydrogenase, we suspect that the oxygen sensor associated with oxytropism does not control the induction of hypoxic metabolism. Our results indicate that oxygen can play a critical role in determining root orientation as well as impacting root metabolic status. Oxytropism allows roots to avoid oxygen-deprived soil strata and may also be the basis of an auto-avoidance mechanism, decreasing the competition between roots for water and nutrients as well as oxygen.

  2. The tropic response of plant roots to oxygen: oxytropism in Pisum sativum L

    NASA Technical Reports Server (NTRS)

    Porterfield, D. M.; Musgrave, M. E.

    1998-01-01

    Plant roots are known to orient growth through the soil by gravitropism, hydrotropism, and thigmotropism. Recent observations of plant roots that developed in a microgravity environment in space suggested that plant roots may also orient their growth toward oxygen (oxytropism). Using garden pea (Pisum sativum L. cv. Weibul's Apollo) and an agravitropic mutant (cv. Ageotropum), root oxytropism was studied in the controlled environment of a microrhizotron. A series of channels in the microrhizotron allowed establishment of an oxygen gradient of 0.8 mmol mol-1 mm-1. Curvature of seedling roots was determined prior to freezing the roots for subsequent spectrophotometric determinations of alcohol dehydrogenase activity. Oxytropic curvature was observed all along the gradient in both cultivars of pea. The normal gravitropic cultivar showed a maximal curvature of 45 degrees after 48 h, while the agravitropic mutant curved to 90 degrees. In each cultivar, the amount of curvature declined as the oxygen concentration decreased, and was linearly related to the root elongation rate. Since oxytropic curvature occurred in roots exposed to oxygen concentrations that were not low enough to induce the hypoxically responsive protein alcohol dehydrogenase, we suspect that the oxygen sensor associated with oxytropism does not control the induction of hypoxic metabolism. Our results indicate that oxygen can play a critical role in determining root orientation as well as impacting root metabolic status. Oxytropism allows roots to avoid oxygen-deprived soil strata and may also be the basis of an auto-avoidance mechanism, decreasing the competition between roots for water and nutrients as well as oxygen.

  3. Callose deposition during gravitropism of Zea mays and Pisum sativum and its inhibition by 2-deoxy-D-glucose

    NASA Technical Reports Server (NTRS)

    Jaffe, M. J.; Leopold, A. C.

    1984-01-01

    In etiolated corn (Zea mays L.) and etiolated pea (Pisum sativum L.) seedlings, a gravitropic stimulation induces the deposition of callose. In the corn coleoptiles this occurs within 5 min of gravity stimulation, and prior to the beginning of curvature. Both gravitropic curvature and callose deposition reach their maxima by 12 h. Within the first 2 h more callose is deposited on the upper (concave) side, but after 2-3 h, this deposition pattern is reversed. An inhibitor of protein glycosylation, 2-deoxy-D-glucose (DDG), inhibits callose production and considerably retards gravitropic bending in both species of plants. Mannose can relieve the inhibition of gravitropic bending by DDG. The pea mutant "Ageotropum", which does not respond to gravity when etiolated, also fails to produce callose in response to a gravitic stimulus. These correlations indicate that callose deposition may be a biochemical component of gravitropism in plant shoots.

  4. Callose deposition during gravitropism of Zea mays and Pisum sativum and its inhibition by 2-deoxy-D-glucose

    NASA Technical Reports Server (NTRS)

    Jaffe, M. J.; Leopold, A. C.

    1984-01-01

    In etiolated corn (Zea mays L.) and etiolated pea (Pisum sativum L.) seedlings, a gravitropic stimulation induces the deposition of callose. In the corn coleoptiles this occurs within 5 min of gravity stimulation, and prior to the beginning of curvature. Both gravitropic curvature and callose deposition reach their maxima by 12 h. Within the first 2 h more callose is deposited on the upper (concave) side, but after 2-3 h, this deposition pattern is reversed. An inhibitor of protein glycosylation, 2-deoxy-D-glucose (DDG), inhibits callose production and considerably retards gravitropic bending in both species of plants. Mannose can relieve the inhibition of gravitropic bending by DDG. The pea mutant "Ageotropum", which does not respond to gravity when etiolated, also fails to produce callose in response to a gravitic stimulus. These correlations indicate that callose deposition may be a biochemical component of gravitropism in plant shoots.

  5. Stimulation of nodulation in field peas (Pisum sativum) by low concentrations of ammonium in hydroponic culture

    NASA Technical Reports Server (NTRS)

    Waterer, J. G.; Vessey, J. K.; Raper, C. D. Jr; Raper CD, J. r. (Principal Investigator)

    1992-01-01

    Although the inhibitory effects of high concentrations of mineral N (> 1.0 mM) on nodule development and function have often been studied, the effects of low, static concentrations of NH4+ (< 1.0 mM) on nodulation are unknown. In the present experiments we examine the effects of static concentrations of NH4+ at 0, 0.1 and 0.5 mM in flowing, hydroponic culture on nodule establishment and nitrogenase activity in field peas [Pisum sativum L. cv. Express (Svalof AB)] for the initial 28 days after planting (DAP). Peas grown in the presence of low concentrations of NH4+ had significantly greater nodule numbers (up to 4-fold) than plants grown without NH4+. Nodule dry weight per plant was significantly higher at 14, 21 and 28 DAP in plants grown in the presence of NH4+, but individual nodule mass was lower than in plants grown without NH4+. The nodulation pattern of the plants supplied with NH4+ was similar to that often reported for supernodulating mutants, however the plants did not express other growth habits associated with supernodulation. Estimates of N2 fixation indicate that the plus-NH4+ peas fixed as much or more N2 than the plants supplied with minus-NH4+ nutrient solution. There were no significant differences in nodule numbers, nodule mass or NH4+ uptake between the plants grown at the two concentrations of NH4+. Nodulation appeared to autoregulate by 14 DAP in the minus-NH4+ treatment. Plant growth and N accumulation in the minus-NH4+ plants lagged behind those of the plus-NH4+ treatments prior to N2 fixation becoming well established in the final week of the experiment. The plus-NH4+ treatments appeared not to elicit autoregulation and plants continued to initiate nodules throughout the experiment.

  6. Stimulation of nodulation in field peas (Pisum sativum) by low concentrations of ammonium in hydroponic culture

    NASA Technical Reports Server (NTRS)

    Waterer, J. G.; Vessey, J. K.; Raper, C. D. Jr; Raper CD, J. r. (Principal Investigator)

    1992-01-01

    Although the inhibitory effects of high concentrations of mineral N (> 1.0 mM) on nodule development and function have often been studied, the effects of low, static concentrations of NH4+ (< 1.0 mM) on nodulation are unknown. In the present experiments we examine the effects of static concentrations of NH4+ at 0, 0.1 and 0.5 mM in flowing, hydroponic culture on nodule establishment and nitrogenase activity in field peas [Pisum sativum L. cv. Express (Svalof AB)] for the initial 28 days after planting (DAP). Peas grown in the presence of low concentrations of NH4+ had significantly greater nodule numbers (up to 4-fold) than plants grown without NH4+. Nodule dry weight per plant was significantly higher at 14, 21 and 28 DAP in plants grown in the presence of NH4+, but individual nodule mass was lower than in plants grown without NH4+. The nodulation pattern of the plants supplied with NH4+ was similar to that often reported for supernodulating mutants, however the plants did not express other growth habits associated with supernodulation. Estimates of N2 fixation indicate that the plus-NH4+ peas fixed as much or more N2 than the plants supplied with minus-NH4+ nutrient solution. There were no significant differences in nodule numbers, nodule mass or NH4+ uptake between the plants grown at the two concentrations of NH4+. Nodulation appeared to autoregulate by 14 DAP in the minus-NH4+ treatment. Plant growth and N accumulation in the minus-NH4+ plants lagged behind those of the plus-NH4+ treatments prior to N2 fixation becoming well established in the final week of the experiment. The plus-NH4+ treatments appeared not to elicit autoregulation and plants continued to initiate nodules throughout the experiment.

  7. Efficient intergeneric fusion of pea (Pisum sativum L.) and grass pea (Lathyrus sativus L.) protoplasts.

    PubMed

    Durieu, P; Ochatt, S J

    2000-07-01

    Large numbers of viable protoplasts of pea (Pisum sativum) and grass pea (Lathyrus sativus) were efficiently and reproducibly obtained and, for the first time, fused. Different procedures for fusion were compared, based either on electrofusion (750, 1000, 1250 or 1500 V cm(-1)), or on the use of macro or micromethods with a polyethylene glycol (PEG 6000 or PEG 1540), or a glycine/high pH solution. Over 10% of viable heterokaryons were obtained, with PEG as the most efficient and reproducible agent for protoplast fusion (>20% of viable heterokaryons). Both the division of heterokaryons and the formation of small calluses were observed.

  8. Phosphatidylinositol(4,5)bisphosphate and phosphatidylinositol(4)phosphate in plant tissues. [Pisum sativum

    SciTech Connect

    Irvine, R.F.; Letcher, A.J.; Lander, D.J. ); Dawson, A.P. ); Musgrave, A. ); Drobak, B.K. )

    1989-03-01

    Pea (Pisum sativum) leaf discs or swimming suspensions of Chlamydomonas eugametos were radiolabeled with ({sup 3}H)myo-inositol or ({sup 32}P)Pi and the lipids were extracted, deacylated, and their glycerol moieties removed. The resulting inositol trisphosphate and bisphosphate fractions were examined by periodate degradation, reduction and dephosphorylation, or by incubation with human red cell membranes. Their likely structures were identified as D-myo-inositol(1,4,5)trisphosphate and D-myo-inositol(1,4,)-bisphosphate. It is concluded that plants contain phosphatidylinositol(4)phosphate and phosphatidylinositol(4,5)bisphosphate; no other polyphosphoinositides were detected.

  9. Exposure of Vicia faba and Pisum sativum to copper-induced genotoxicity.

    PubMed

    Souguir, D; Ferjani, E; Ledoigt, G; Goupil, P

    2008-11-01

    The potential genotoxicity of Cu(2+) was investigated in Vicia faba and Pisum sativum seedlings in hydroponic culture conditions. Cu(2+) caused a dose-dependent increase in micronuclei frequencies in both plant models. Cytological analysis of root tips cells showed clastogenic and aneugenic effects of this heavy metal on V. faba root meristems. Cu(2+) induced chromosomal alterations at the lowest concentration used (2.5 mM) when incubated for 42 h, indicating the potent mutagenic effect of this ion. A spectrum of chromosomal abnormalities was observed in V. faba root meristems, illustrating the genotoxic events leading to micronuclei formation.

  10. Essentiality of Boron for Symbiotic Dinitrogen Fixation in Pea (Pisum sativum) Rhizobium Nodules.

    PubMed Central

    Bolanos, L.; Esteban, E.; De Lorenzo, C.; Fernandez-Pascual, M.; De Felipe, M. R.; Garate, A.; Bonilla, I.

    1994-01-01

    The effect of boron deficiency on symbiotic nitrogen fixation in pea (Pisum sativum) was examined. The absence of boron in the culture medium resulted in a decrease of the number of nodules and an alteration of nodule development leading to an inhibition of nitrogenase activity. Examination of boron-deficient nodules showed dramatic changes in cell walls and in both peribacteroid and infection thread membranes, suggesting a role for boron in the stability of these structures. These results indicate that boron is a requirement for normal nodule development and functionality. PMID:12232064

  11. Fucosylation of xyloglucan: localization of the transferase in dictyosomes of pea stem cells. [Pisum sativum

    SciTech Connect

    Camirand, A.; Brummell, D.; MacLachlan, G.

    1987-07-01

    Microsomal membranes from elongating regions of etiolated Pisum sativum stems were separated by rate-zonal centrifugation on Renografin gradients. The transfer of labeled fucose and xylose from GDP-(/sup 14/C) fucose and UDP-(/sup 14/C)xylose to xyloglucan occurred mainly in dictyosome-enriched fractions. No transferase activity was detected in secretory vesicle fractions. Pulse-chase experiments using pea stem slices incubated with (/sup 3/H)fucose suggest that xyloglucan chains are fucosylated and their structure completed within the dictyosomes, before being transported to the cell wall by secretory vesicles.

  12. Extracellular superoxide production associated with secondary root growth following desiccation of Pisum sativum seedlings.

    PubMed

    Roach, Thomas; Kranner, Ilse

    2011-10-15

    The seedling stage is arguably the most vulnerable phase in the plant life cycle, where the young establishing plant is extremely sensitive to environmental stresses such as drought. Here, the production of superoxide (O(2)(-)), a molecule involved in stress signaling, was measured in response to desiccation of Pisum sativum L. seedlings. Following desiccation that was sufficient to kill the radicle meristem, viability could be retained by seedlings that grew secondary roots. Upon rehydration, secondary roots formed in a region that had displayed intense extracellular O(2)(-)production on desiccation. Treating partially desiccated seedlings with hydrogen peroxide (H(2)O(2)) prevented viability loss. In summary, reactive oxygen species (ROS) appear to participate in the signaling required for secondary root formation following desiccation stress of P. sativum seedlings.

  13. The same allele of translation initiation factor 4E mediates resistance against two Potyvirus spp. in Pisum sativum.

    PubMed

    Bruun-Rasmussen, M; Møller, I S; Tulinius, G; Hansen, J K R; Lund, O S; Johansen, I E

    2007-09-01

    Pathogenicity of two sequenced isolates of Bean yellow mosaic virus (BYMV) was established on genotypes of Pisum sativum L. reported to carry resistance genes to BYMV and other potyviruses. Resistance to the white lupin strain of BYMV (BYMV-W) is inherited as a recessive gene named wlv that maps to linkage group VI together with other Potyvirus resistances. One of these, sbm1, confers resistance to strains of Pea seedborne mosaic virus and previously has been identified as a mutant allele of the eukaryotic translation initiation factor 4E gene (eIF4E). Sequence comparison of eIF4E from BYMV-W-susceptible and -resistant P. sativum genotypes revealed a polymorphism correlating with the resistance profile. Expression of eIF4E from susceptible plants in resistant plants facilitated BYMV-W infection in inoculated leaves. When cDNA of BYMV-W was agroinoculated, resistance mediated by the wlv gene frequently was overcome, and virus from these plants had a codon change causing an Arg to His change at position 116 of the predicted viral genome-linked protein (VPg). Accordingly, plants carrying the wlv resistance gene were infected upon inoculation with BYMV-W derived from cDNA with a His codon at position 116 of the VPg coding region. These results suggested that VPg determined pathogenicity on plants carrying the wlv resistance gene and that wlv corresponded to the sbm1 allele of eIF4E.

  14. Growth and some physiological attributes of pea (Pisum sativum L.) as affected by salinity.

    PubMed

    Najafi, F; Khavari-Nejad, R A; Rastgar-Jazii, F; Sticklen, M

    2007-08-15

    The effects of salt stress were studied on growth and physiology of pea (Pisum sativum L. cv. Green Arrow) in a pot study. Pea plants were treated with NaCl at 0, 10, 30, 50 and 70 mM in Hoagland solution. Plants were harvested after 21 days for measurements of physiological parameters. The highest NAR and RGR were found in 10 mM NaCl. However, in 70 mM NaCl, RGR and RLGR were significantly decreased in respect of other concentrations of NaCl. In 50 and 70 mM NaCl, chlorophylls contents and photosynthetic rate, were significantly decreased and CO2 compensation concentration and respiration rate increased in comparison with control. In 10 and 30 mM NaCl gas exchanges and chlorophyll contents were not significantly decrease in respect of control. Results indicated that Pisum sativum L. cv. Green Arrow can tolerate to 70 mM NaCl, also growth of plants in 10 and 30 mM NaCl was better than that of those in 0 mM NaCl.

  15. Rhizobium anhuiense sp. nov., isolated from effective nodules of Vicia faba and Pisum sativum.

    PubMed

    Zhang, Yu Jing; Zheng, Wen Tao; Everall, Isobel; Young, J Peter W; Zhang, Xiao Xia; Tian, Chang Fu; Sui, Xin Hua; Wang, En Tao; Chen, Wen Xin

    2015-09-01

    Four rhizobia-like strains, isolated from root nodules of Pisum sativum and Vicia faba grown in Anhui and Jiangxi Provinces of China, were grouped into the genus Rhizobium but were distinct from all recognized species of the genus Rhizobium by phylogenetic analysis of 16S rRNA and housekeeping genes. The combined sequences of the housekeeping genes atpD, recA and glnII for strain CCBAU 23252(T) showed 86.9 to 95% similarity to those of known species of the genus Rhizobium. All four strains had nodC and nifH genes and could form effective nodules with Pisum sativum and Vicia faba, and ineffective nodules with Phaseolus vulgaris, but did not nodulate Glycine max, Arachis hypogaea, Medicago sativa, Trifolium repens or Lablab purpureus in cross-nodulation tests. Fatty acid composition, DNA-DNA relatedness and a series of phenotypic tests also separated these strains from members of closely related species. Based on all the evidence, we propose a novel species, Rhizobium anhuiense sp. nov., and designate CCBAU 23252(T) ( = CGMCC 1.12621(T) = LMG 27729(T)) as the type strain. This strain was isolated from a root nodule of Vicia faba and has a DNA G+C content of 61.1 mol% (Tm).

  16. Association mapping of yield candidate gene homologs in a diverse collection of pea (Pisum sativum L.) lines

    USDA-ARS?s Scientific Manuscript database

    Association mapping, based on linkage disequilibrium (LD), is used increasingly to describe associations between allelic variation and phenotype. Yield is a key economic trait for most field crops, including pea (Pisum sativum L.). Recent reports in plant systems have identified candidate genes fo...

  17. Induction of Glutamine Synthetase Activity in Nonnodulated Roots of Glycine max, Phaseolus vulgaris, and Pisum sativum1

    PubMed Central

    Hoelzle, Inger; Finer, John J.; McMullen, Michael D.; Streeter, John G.

    1992-01-01

    Nitrate or ammonium fertilization significantly increased glutamine synthetase (GS) activity in nonnodulated roots of French bean (Phaseolus vulgaris), soybean (Glycine max), and pea (Pisum sativum). Western analysis revealed substantial GS antibody-positive protein in root extracts that had minimal GS activity, indicating that an inactive form of GS may be present in nonfertilized plants. Images Figure 1 PMID:16652993

  18. Effect of clinorotation on the leaf mesophyll structure and pigment content in Arabidopsis thaliana L. and Pisum sativum L.

    PubMed

    Adamchuk, N I

    2004-07-01

    Properties of mesophyll cells and photosynthetic membranes of Arabidopsis thaliana (L.) Heynh. and Pisum sativum (L.) plants grown in a horizontal clinostat and in control conditions were compared. Obtained data have show that under clinorotation conditions seedlings have experienced the following cell morphology changes structural chloroplast rearrangement in palisade cells, pigment content alteration, and cell aging acceleration.

  19. Products of dark CO sub 2 fixation in pea root nodules support bacteroid metabolism. [Pisum sativum L

    SciTech Connect

    Rosendahl, L.; Pedersen, W.B. ); Vance, C.P. )

    1990-05-01

    Products of the nodule cytosol in vivo dark ({sup 14}C)CO{sub 2} fixation were detected in the plant cytosol as well as in the bacteroids of pea (Pisum sativum L. cv Bodil) nodules. The distribution of the metabolites of the dark CO{sub 2} fixation products was compared in effective (fix{sup +}) nodules infected by a wild-type Rhizobium leguminosarum (MNF 300), and ineffective (fix{sup {minus}}) nodules of the R. leguminosarum mutant MNF 3080. The latter has a defect in the dicarboxylic acid transport system of the bacterial membrane. The {sup 14}C incorporation from ({sup 14}C)CO{sub 2} was about threefold greater in the wild-type nodules than in the mutant nodules. Similarly, in wild-type nodules the in vitro phosphoenolpyruvate carboxylase activity was substantially greater than that of the mutant. Almost 90% of the {sup 14}C label in the cytosol was found in organic acids in both symbioses. The results indicate a central role for nodule cytosol dark CO{sub 2} fixation in the supply of the bacteroids with dicarboxylic acids.

  20. Differential induction of Pisum sativum defense signaling molecules in response to pea aphid infestation.

    PubMed

    Mai, Van Chung; Drzewiecka, Kinga; Jeleń, Henryk; Narożna, Dorota; Rucińska-Sobkowiak, Renata; Kęsy, Jacek; Floryszak-Wieczorek, Jolanta; Gabryś, Beata; Morkunas, Iwona

    2014-05-01

    This study demonstrates the sequence of enhanced generation of signal molecules such as phytohormones, i.e. jasmonic acid (JA), ethylene (ET), salicylic acid (SA), and a relatively stable free radical, nitric oxide (NO), in response of Pisum sativum L. cv. Cysterski seedling leaves to the infestation of pea aphid Acyrthosiphon pisum (Harris) at a varied population size. In time from 0 to 96h after A. pisum infestation these signal molecules accumulated transiently. Moreover, the convergence of these signaling pathways occurred. JA and its methyl derivative MeJA reached the first maximum of generation at 24th hour of infestation. An increase in ET and NO generation was observed at 48th hour of infestation. The increase in SA, JA/MeJA and ET concentrations in aphid-infested leaves occurred from the 72nd to 96th hour. In parallel, an increase was demonstrated for the activities of enzymes engaged in the biosynthesis of SA, such as phenylalanine ammonia-lyase (PAL) and benzoic acid 2-hydroxylase (BA2H). Additionally, a considerable post-infestation accumulation of transcripts for PAL was observed. An increase in the activity of lipoxygenase (LOX), an important enzyme in the biosynthesis of JA was noted. This complex signaling network may contribute to the coordinated regulation of gene expression leading to specific defence responses. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  1. Assessment of genetic diversity in Ethiopian field pea (Pisum sativum L.) accessions with newly developed EST-SSR markers.

    PubMed

    Teshome, Abel; Bryngelsson, Tomas; Dagne, Kifle; Geleta, Mulatu

    2015-08-19

    Field pea (Pisum sativum L.) is among the prominent crops in the world as food and feed. There are relatively few simple sequence repeat (SSR) markers developed from expressed sequence tags (ESTs) in P. sativum. In the present study, 15 new EST-SSR markers were developed from publicly available ESTs. These markers have successfully amplified their target loci across seven Pisum sativum subsp. sativum accessions. Eleven (73%) of these SSRs were trinucleotide repeats, two (13%) dinucleotide and two (13%) were hexanucleotide repeats. Across-taxa transferability of these new markers was also tested on other subspecies of Pisum as well as on P. fulvum, Vicia faba and Lens culinaris. In Pisum sativum subsp. sativum, 13 of the 15 markers were polymorphic and 12 of them subsequently used for genetic diversity analysis. Forty six accessions, of which 43 were from Ethiopia, were subjected to genetic diversity analysis using these newly developed markers. All accessions were represented by 12 individuals except two (NGB103816 and 237508) that were represented by 9 and 11 individuals, respectively. A total of 37 alleles were detected across all accessions. PS10 was the most polymorphic locus with six alleles, and the average number of alleles per locus over the 12 polymorphic loci was 3.1. Several rare and private alleles were also revealed. The most distinct accession (32048) had private alleles at three loci with 100% frequency. These newly developed EST-SSR primer-pairs successfully amplified expected loci in P. sativum subsp. sativum as well as in other subspecies of the genus Pisum and related genera. High levels of genetic variation were detected in field pea accessions from Ethiopia using these markers. This result implies the potential of the Ethiopian field pea gene pool for improvement of field peas in various desirable traits. In addition, these markers could be a valuable asset in resolving the inconsistency in the taxonomic status of the different subspecies of

  2. Imperative roles of salicylic acid and nitric oxide in improving salinity tolerance in Pisum sativum L.

    PubMed

    Yadu, Shrishti; Dewangan, Teman Lal; Chandrakar, Vibhuti; Keshavkant, S

    2017-01-01

    This study was undertaken to scrutinize efficacy of salicylic acid (SA) and/or sodium nitroprusside [SNP, source of nitric oxide (NO)] to mitigate injury symptoms of saline stress in Pisum sativum L. Exposure to sodium chloride (NaCl) was found to be injurious to germinating P. sativum L. (var. Shubhra IM-9101) and a direct correlation between severity of toxicity and NaCl-concentrations could be discernible. Both SA and NO serves as signal molecules in plant stress responses, and play crucial roles in key regulatory pathways of growth, development and metabolism. The limiting effects of salinity on radicle length and biomass accumulation were considerably released by SA and/or SNP and among which their combined application was found to be the most promising. Supplemented SA and/or SNP, particularly their cocktail, resulted in a substantial decline in reactive oxygen species accumulation, which later caused reduced accumulations of malondialdehyde, 4-hydroxy-2-nonenal and protein carbonyl, in NaCl subjected germinating P. sativum L. seeds. SA and/or SNP had significant inducing effects on activities of superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase. Additionally, exogenous SA and/or SNP led to the higher proline, sugar and glycinebetaine contents, than that of the control. On the basis of accumulated results, it could be concluded that the cocktail of SA and SNP may be efficiently used to overcome the adverse signatures of salinity stress.

  3. SGRL can regulate chlorophyll metabolism and contributes to normal plant growth and development in Pisum sativum L.

    PubMed

    Bell, Andrew; Moreau, Carol; Chinoy, Catherine; Spanner, Rebecca; Dalmais, Marion; Le Signor, Christine; Bendahmane, Abdel; Klenell, Markus; Domoney, Claire

    2015-12-01

    Among a set of genes in pea (Pisum sativum L.) that were induced under drought-stress growth conditions, one encoded a protein with significant similarity to a regulator of chlorophyll catabolism, SGR. This gene, SGRL, is distinct from SGR in genomic location, encoded carboxy-terminal motif, and expression through plant and seed development. Divergence of the two encoded proteins is associated with a loss of similarity in intron/exon gene structure. Transient expression of SGRL in leaves of Nicotiana benthamiana promoted the degradation of chlorophyll, in a manner that was distinct from that shown by SGR. Removal of a predicted transmembrane domain from SGRL reduced its activity in transient expression assays, although variants with and without this domain reduced SGR-induced chlorophyll degradation, indicating that the effects of the two proteins are not additive. The combined data suggest that the function of SGRL during growth and development is in chlorophyll re-cycling, and its mode of action is distinct from that of SGR. Studies of pea sgrL mutants revealed that plants had significantly lower stature and yield, a likely consequence of reduced photosynthetic efficiencies in mutant compared with control plants under conditions of high light intensity.

  4. Malate Dehydrogenases of Pisum sativum: Tissue Distribution and Properties of the Particulate Forms.

    PubMed

    Zschoche, W C; Ting, I P

    1973-06-01

    Mitochondria and leaf microbodies isolated from leaves of pea (Pisum sativum) by sucrose density gradient centrifugation were each shown to have a unique form (isoenzyme) of malate dehydrogenase (EC 1.1.1.37) based on chromatographic and kinetic properties. Root organelle preparations were shown to contain only a mitochondrial malate dehydrogenase with physical and kinetic properties similar to the leaf form. The absence of a detectable root microbody malate dehydrogenase similar to the leaf enzyme, which is intermediate in electrophoretic and chromatographic properties between the mitochondrial and soluble isoenzymes, was confirmed by diethylaminoethyl cellulose column chromatography and starch-gel electrophoresis of total homogenates from leaf and root tissue. These findings tend to support the role of the leaf microbody isoenzyme in a pathway unique to photosynthetic tissue.

  5. A glucuronyltransferase involved in glucuronoxylan synthesis in pea (Pisum sativum) epicotyls.

    PubMed Central

    Waldron, K W; Brett, C T

    1983-01-01

    A particulate enzyme preparation made from epicotyls of 1-week-old etiolated pea (Pisum sativum) seedlings was shown to incorporate glucuronic acid from UDP-D-[U-14C]glucuronic acid into a hemicellulosic polysaccharide. Optimum conditions for the incorporation include the presence of Mn2+ ions at between 4 and 10 mmol/litre and a pH between 5 and 6. UDP-D-xylose at 1 mmol/litre allows incorporation to continue for at least 8 h. In its absence, the reaction stops within 30 min. Analysis of the product by partial and total acid hydrolysis, followed by paper chromatography or electrophoresis, indicates that the polysaccharide produced is a glucuronoxylan. PMID:6412678

  6. Effect of Ethylene on Cell Division and Deoxyribonucleic Acid Synthesis in Pisum sativum1

    PubMed Central

    Apelbaum, Akiva; Burg, Stanley P.

    1972-01-01

    Ethylene and supraoptimal levels of 2,4-dichlorophenoxyacetic acid inhibit the growth of the apical hook region of etiolated Pisum sativum (var. Alaska) seedlings by stopping almost all cell divisions. Cells are prevented from entering prophase. The hormones also retard cell division in intact root tips and completely stop the process in lateral buds. The latter inhibition is reversed partially by benzyl adenine. In root tips and the stem plumular and subhook regions, ethylene inhibits DNA synthesis. The magnitude of this inhibition is correlated with the degree of repression of cell division in meristematic tissue, suggesting that the effect on cell division results from a lack of DNA synthesis. Ethylene inhibits cell division within a few hours with a dose-response curve similar to that for most other actions of the gas. Experiments with seedlings grown under hypobaric conditions suggest that the gas naturally controls plumular expansion and cell division in the apical region. Images PMID:16658105

  7. Rapidly Induced Wound Ethylene from Excised Segments of Etiolated Pisum sativum L., cv. Alaska

    PubMed Central

    Saltveit, Mikal E.; Dilley, David R.

    1978-01-01

    Increased ethylene synthesis was rapidly induced throughout the apical meristematic region of etiolated seedlings of Pisum sativum L., cv. Alaska by cuts made 1 centimeter from the apical hook. The wound signal was transmitted at about 2 millimeters per minute. Accumulation of substance(s) at the cut surfaces of excised sections, as the result of interrupted translocation, did not initiate or significantly contribute to wound-induced ethylene synthesis, nor was the cut surface the site of enhanced ethylene synthesis. Cutting subapical sections into shorter pieces showed that cells less than 2 millimeters from a cut surface produced about 30% less ethylene than cells greater than 2 millimeters from a cut surface. PMID:16660590

  8. [Meiotic abnormalities as expression of nuclear-cytoplasmic incompatibility in crosses of Pisum sativum subspecies].

    PubMed

    Bogdanova, V S; Galieva, E R

    2009-05-01

    Meiosis in anthers and mitosis in somatic cells were studied in reciprocal F1 hybrids of the accession VIR320, which belonged to wild Pisum sativum ssp. elatius (Bieb.) Schmal., and the laboratory line Sprint-1. When VIR320 was used as a maternal form, the hybrids displayed nuclear-cytoplasmic conflict, which caused chlorophyll defects and meiotic abnormalities. One or two chromosomes lagged in the equatorial region during chromosome segregation to the poles, distorting cytokinesis and yielding abnormal microspores. Chlorophyll defects were not observed, and meiotic abnormalities were far less frequent in reciprocal hybrids and in the case of an abnormal paternal inheritance of plastids from Sprint-1. Mitosis lacked overt abnormalities in all of the hybrids.

  9. Photosynthesis and growth responses of pea Pisum sativum L. under heavy metals stress.

    PubMed

    Hattab, Sabrine; Dridi, Boutheina; Chouba, Lassad; Ben, Kheder Mohamed; Bousetta, Hamadi

    2009-01-01

    The present work aimed to study the physiological effects of cadmium (Cd) and copper (Cu) in pea (Pisum sativum). Pea plants were exposed to increasing doses of cadmium chloride (CdCl2) and copper chloride (CuCl2) for 20 d. The examined parameters, namely root and shoot lengths, the concentration of photosynthetic pigments and the rate of photosynthesis were affected by the treatments especially with high metals concentrations. The analysis of heavy metals accumulation shows that leaves significantly accumulate cadmium for all the tested concentrations. However, copper was significantly accumulated only with the highest tested dose. This may explain the higher inhibitory effects of cadmium on photosynthesis and growth in pea plants. These results are valuable for understanding the biological consequences of heavy metals contamination particularly in soils devoted to organic agriculture.

  10. Nitrogen deficiency hinders etioplast development in stems of dark-grown pea (Pisum sativum) shoot cultures.

    PubMed

    Kósa, Annamária; Preininger, Éva; Böddi, Béla

    2015-11-01

    The effects of nitrogen (N) deprivation were studied in etiolated pea plants (Pisum sativum cv. Zsuzsi) grown in shoot cultures. The average shoot lengths decreased and the stems significantly altered considering their pigment contents, 77 K fluorescence spectra and ultrastructural properties. The protochlorophyllide (Pchlide) content and the relative contribution of the 654-655 nm emitting flash-photoactive Pchlide form significantly decreased. The etioplast inner membrane structure characteristically changed: N deprivation correlated with a decrease in the size and number of prolamellar bodies (PLBs). These results show that N deficiency directly hinders the pigment production, as well as the synthesis of other etioplast inner membrane components in etiolated pea stems. © 2015 Scandinavian Plant Physiology Society.

  11. Development of an indirect enzyme linked immunoassay for abscisic acid. [Pisum sativum

    SciTech Connect

    Ross, G.S.; Elder, P.A.; McWha, J.A.; Pearce, D.; Pharis, R.P.

    1987-09-01

    AN INDIRECT METHOD OF ENZYME-LINKED-IMMUNOSORBENT-ASSAY (ELISA) IS REPORTED FOR ABSCISIC ACID (ABA), UTILIZING A THYROGLOBULIN-ABA CONJUGATE FOR COATING WELLS. THE ASSAY CAN USE COMMERCIALLY AVAILABLE MONOCLONAL ANTIBODIES, IS SENSITIVE TO AS LITTLE AS 20 PICOGRAMS ABA PER WELL, AND IS MUCH MORE CONSERVATIVE OF ANTIBODY THAN DIRECT METHODS. THE MOST DILUTE ABA STANDARDS DID NOT RETAIN THEIR ANTIGENICITY DURING STORAGE, SO ABA STANDARD SETS WERE DILUTED IMMEDIATELY PRIOR TO USE. THE INDIRECT ELISA WAS USED SUCCESSFULLY TO ESTIMATE ABA CONCENTRATIONS IN DEVELOPING COTYLEDONS OF PISUM SATIVUM L., AFTER ONLY LITTLE PRELIMINARY PURIFICATION. IT WAS VALIDATED FOR THIS TISSUE THROUGH THE USE OF GAS CHROMATOGRAPHY-ELECTRON CAPTURE DETECTION (GC-EC), AND CAPILLARY GC-SELECTED ION MONITORING (GC-MS-SIM) USING LABELLED ABA AS AN INTERNAL STANDARD. FULL SPECTRUM GC-MASS SPECTROMETRY WAS ALSO USED TO VERIFY THAT ABA WAS PRESENT IN A SAMPLE ASSAYED QUANTITATIVELY BY BOTH ELISA AND GC-MS-SIM.

  12. Development and characterization of 37 novel EST-SSR markers in Pisum sativum (Fabaceae).

    PubMed

    Zhuang, Xiaofeng; McPhee, Kevin E; Coram, Tristan E; Peever, Tobin L; Chilvers, Martin I

    2013-01-01

    Simple sequence repeat markers were developed based on expressed sequence tags (EST-SSR) and screened for polymorphism among 23 Pisum sativum individuals to assist development and refinement of pea linkage maps. In particular, the SSR markers were developed to assist in mapping of white mold disease resistance quantitative trait loci. • Primer pairs were designed for 46 SSRs identified in EST contiguous sequences assembled from a 454 pyrosequenced transcriptome of the pea cultivar, 'LIFTER'. Thirty-seven SSR markers amplified PCR products, of which 11 (30%) SSR markers produced polymorphism in 23 individuals, including parents of recombinant inbred lines, with two to four alleles. The observed and expected heterozygosities ranged from 0 to 0.43 and from 0.31 to 0.83, respectively. • These EST-SSR markers for pea will be useful for refinement of pea linkage maps, and will likely be useful for comparative mapping of pea and as tools for marker-based pea breeding.

  13. Root and foliar uptake, translocation, and distribution of [14C] fluoranthene in pea plants (Pisum sativum).

    PubMed

    Zezulka, Stěpán; Klemš, Marek; Kummerová, Marie

    2014-10-01

    Uptake of (14)C-labeled fluoranthene ([(14)C]FLT) via both roots and leaves of Pisum sativum seedlings and distribution of [(14) C] in plants by both acropetal and basipetal transport was evaluated. The highest [(14)C] level was found in the root base (≈270 × 10(4) dpm/g dry wt) and the lowest level in the stem apex (<2 × 10(4) dpm/g dry wt) after just 2 h of root exposure. For foliar uptake, the highest level of [(14)C] was found in the stem and root apex (both ≈2 × 10(4) dpm/g dry wt) (except for treated leaves), while the lowest level was found in the root base (<0.6 × 10(4) dpm/g dry wt). © 2014 SETAC.

  14. On the shock response of Pisum Sativum (a.k.a the Common Pea)

    NASA Astrophysics Data System (ADS)

    Leighs, James; Hazell, Paul; Appleby-Thomas, Gareth

    2011-06-01

    The high strain-rate response of biological and organic structures is of interest to numerous fields ranging from the food industry (dynamic pasteurisation) to astrobiology (e.g. the theory of panspermia, which suggests that planets could be `seeded' with life `piggy-backing' of interplanetary bodies). Consequently, knowledge of the damage mechanisms and viability of shocked organic material is of paramount importance. In this study a single-stage gas-gun has been employed to subject samples of Pisum Sativum (the Common Pea) to semi-planar shock loading, corresponding to impact pressures of up to c.3 GPa. The experimental approach adopted is discussed along with results from Manganin gauges embedded in the target capsule which show the loading history. Further, the viability of the shock-loaded peas was investigated via attempts at germination. Finally, microscopic examination of the impacted specimens allowed a qualitative assessment of damage mechanisms to be made.

  15. Comprehensive transcriptome analysis of the highly complex Pisum sativum genome using next generation sequencing

    PubMed Central

    2011-01-01

    Background The garden pea, Pisum sativum, is among the best-investigated legume plants and of significant agro-commercial relevance. Pisum sativum has a large and complex genome and accordingly few comprehensive genomic resources exist. Results We analyzed the pea transcriptome at the highest possible amount of accuracy by current technology. We used next generation sequencing with the Roche/454 platform and evaluated and compared a variety of approaches, including diverse tissue libraries, normalization, alternative sequencing technologies, saturation estimation and diverse assembly strategies. We generated libraries from flowers, leaves, cotyledons, epi- and hypocotyl, and etiolated and light treated etiolated seedlings, comprising a total of 450 megabases. Libraries were assembled into 324,428 unigenes in a first pass assembly. A second pass assembly reduced the amount to 81,449 unigenes but caused a significant number of chimeras. Analyses of the assemblies identified the assembly step as a major possibility for improvement. By recording frequencies of Arabidopsis orthologs hit by randomly drawn reads and fitting parameters of the saturation curve we concluded that sequencing was exhaustive. For leaf libraries we found normalization allows partial recovery of expression strength aside the desired effect of increased coverage. Based on theoretical and biological considerations we concluded that the sequence reads in the database tagged the vast majority of transcripts in the aerial tissues. A pathway representation analysis showed the merits of sampling multiple aerial tissues to increase the number of tagged genes. All results have been made available as a fully annotated database in fasta format. Conclusions We conclude that the approach taken resulted in a high quality - dataset which serves well as a first comprehensive reference set for the model legume pea. We suggest future deep sequencing transcriptome projects of species lacking a genomics backbone will

  16. Rubisco activity in guard cells compared with the solute requirement for stomatal opening. [Pisum sativum

    SciTech Connect

    Reckmann, U.; Scheibe, R.; Raschke, K. )

    1990-01-01

    We investigated whether the reductive pentose phosphate path in guard cells of Pisum sativum had the capacity to contribute significantly to the production of osmotica during stomatal opening in the light. Amounts of ribulose 1,5-bisphophate carboxylase/oxygenase (Rubisco) were determined by the ({sup 14}C) carboxyarabinitol bisphosphate assay. A guard cell contained about 1.2 and a mesophyll cell about 324 picograms of the enzyme; the ratio was 1:270. The specific activities of Rubisco in guard cells and in mesophyll cells were equal; there was no indication of a specific inhibitor of Rubisco in guard cells. Rubisco activity was 115 femtomol per guard-cell protoplast and hour. This value was different from zero with a probability of 0.99. After exposure of guard-cell protoplasts to {sup 14}CO{sub 2} for 2 seconds in the light, about one-half of the radioactivity was in phosphorylated compounds and <10% in malate. Guard cells in epidermal strips produced a different labelling pattern; in the light, <10% of the label was in phosphorylated compounds and about 60% in malate. The rate of solute accumulation in intact guard cells was estimated to have been 900 femto-osmol per cell and hour. If Rubisco operated at full capacity in guard cells, and hexoses were produced as osmotica, solutes could be supplied at a rate of 19femto-osmol per cell and hour, which would constitute 2% of the estimated requirement. The capacity of guard-cell Rubisco to meet the solute requirement for stomatal opening in leaves of Pisum sativum is insignificant.

  17. Extraction, purification, kinetic and thermodynamic properties of urease from germinating Pisum Sativum L. seeds.

    PubMed

    El-Hefnawy, Mohamed E; Sakran, Mohamed; Ismail, Ali I; Aboelfetoh, Eman Fahmy

    2014-07-28

    Urease, one of the highly efficient known enzymes, catalyzes the hydrolysis of urea into ammonia and carbon dioxide. The present study aimed to extract urease from pea seeds (Pisum Sativum L). The enzyme was then purified in three consequence steps: acetone precipitation, DEAE-cellulose ion-exchange chromatography, and gel filtration chromatography (Sephacryl S-200 column). The purification fold was 12.85 with a yield of 40%. The molecular weight of the isolated urease was estimated by chromatography to be 269,000 Daltons. Maximum urease activity (190 U/g) was achieved at the optimum conditions of 40°C and pH of 7.5 after 5 min of incubation. The kinetic parameters, Km and Vmax, were estimated by Lineweaver-Burk fits and found to be 500 mM and 333.3 U/g, respectively. The thermodynamic constants of activation, ΔH, Ea, and ΔS, were determined using Arrhenius plot and found to be 21.20 kJ/mol, 23.7 kJ/mol, and 1.18 kJ/mol/K, respectively. Urease was purified from germinating Pisum Sativum L. seeds. The purification fold, yield, and molecular weight were determined. The effects of pH, concentration of enzyme, temperature, concentration of substrate, and storage period on urease activity were examined. This may provide an insight on the various aspects of the property of the enzyme. The significance of extracting urease from different sources could play a good role in understanding the metabolism of urea in plants.

  18. Extraction, purification, kinetic and thermodynamic properties of urease from germinating Pisum Sativum L. seeds

    PubMed Central

    2014-01-01

    Background Urease, one of the highly efficient known enzymes, catalyzes the hydrolysis of urea into ammonia and carbon dioxide. The present study aimed to extract urease from pea seeds (Pisum Sativum L). The enzyme was then purified in three consequence steps: acetone precipitation, DEAE-cellulose ion-exchange chromatography, and gel filtration chromatography (Sephacryl S-200 column). Results The purification fold was 12.85 with a yield of 40%. The molecular weight of the isolated urease was estimated by chromatography to be 269,000 Daltons. Maximum urease activity (190 U/g) was achieved at the optimum conditions of 40°C and pH of 7.5 after 5 min of incubation. The kinetic parameters, K m and V max , were estimated by Lineweaver-Burk fits and found to be 500 mM and 333.3 U/g, respectively. The thermodynamic constants of activation, ΔH, E a , and ΔS, were determined using Arrhenius plot and found to be 21.20 kJ/mol, 23.7 kJ/mol, and 1.18 kJ/mol/K, respectively. Conclusions Urease was purified from germinating Pisum Sativum L. seeds. The purification fold, yield, and molecular weight were determined. The effects of pH, concentration of enzyme, temperature, concentration of substrate, and storage period on urease activity were examined. This may provide an insight on the various aspects of the property of the enzyme. The significance of extracting urease from different sources could play a good role in understanding the metabolism of urea in plants. PMID:25065975

  19. Comparative analysis of the tubulin cytoskeleton organization in nodules of Medicago truncatula and Pisum sativum: bacterial release and bacteroid positioning correlate with characteristic microtubule rearrangements.

    PubMed

    Kitaeva, Anna B; Demchenko, Kirill N; Tikhonovich, Igor A; Timmers, Antonius C J; Tsyganov, Viktor E

    2016-04-01

    In this study we analyzed and compared the organization of the tubulin cytoskeleton in nodules of Medicago truncatula and Pisum sativum. We combined antibody labeling and green fluorescent protein tagging with laser confocal microscopy to observe microtubules (MTs) in nodules of both wild-type (WT) plants and symbiotic plant mutants blocked at different steps of nodule development. The 3D MT organization of each histological nodule zone in both M. truncatula and P. sativum is correlated to specific developmental processes. Endoplasmic MTs appear to support infection thread growth, infection droplet formation and bacterial release into the host cytoplasm in nodules of both species. No differences in the organization of the MT cytoskeleton between WT and bacterial release mutants were apparent, suggesting both that the phenotype is not linked to a defect in MT organization and that the growth of hypertrophied infection threads is supported by MTs. Strikingly, bacterial release coincides with a change in the organization of cortical MTs from parallel arrays into an irregular, crisscross arrangement. After release, the organization of endoplasmic MTs is linked to the distribution of symbiosomes. The 3D MT organization of each nodule histological zone in M. truncatula and P. sativum was analyzed and linked to specific developmental processes. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

  20. Microbial symbionts affect Pisum sativum proteome and metabolome under Didymella pinodes infection.

    PubMed

    Desalegn, G; Turetschek, R; Kaul, H-P; Wienkoop, S

    2016-06-30

    The long cultivation of field pea led to an enormous diversity which, however, seems to hold just little resistance against the ascochyta blight disease complex. The potential of below ground microbial symbiosis to prime the immune system of Pisum for an upcoming pathogen attack has hitherto received little attention. This study investigates the effect of beneficial microbes on the leaf proteome and metabolome as well as phenotype characteristics of plants in various symbiont interactions (mycorrhiza, rhizobia, co-inoculation, non-symbiotic) after infestation by Didymella pinodes. In healthy plants, mycorrhiza and rhizobia induced changes in RNA metabolism and protein synthesis. Furthermore, metal handling and ROS dampening was affected in all mycorrhiza treatments. The co-inoculation caused the synthesis of stress related proteins with concomitant adjustment of proteins involved in lipid biosynthesis. The plant's disease infection response included hormonal adjustment, ROS scavenging as well as synthesis of proteins related to secondary metabolism. The regulation of the TCA, amino acid and secondary metabolism including the pisatin pathway, was most pronounced in rhizobia associated plants which had the lowest infection rate and the slowest disease progression. A most comprehensive study of the Pisum sativum proteome and metabolome infection response to Didymella pinodes is provided. Several distinct patterns of microbial symbioses on the plant metabolism are presented for the first time. Upon D. pinodes infection, rhizobial symbiosis revealed induced systemic resistance e.g. by an enhanced level of proteins involved in pisatin biosynthesis. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  1. Structure of Pisum sativum Rubisco with bound ribulose 1,5-bisphosphate

    PubMed Central

    Loewen, Peter C.; Didychuk, Allison L.; Switala, Jacek; Perez-Luque, Rosa; Fita, Ignacio; Loewen, Michele C.

    2013-01-01

    The first structure of a ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from a pulse crop is reported. Rubisco was purified from Pisum sativum (garden pea) and diffraction-quality crystals were obtained by hanging-drop vapour diffusion in the presence of the substrate ribulose 1,5-bisphosphate. X-ray diffraction data were recorded to 2.20 Å resolution from a single crystal at the Canadian Light Source. The overall quaternary structure of non-activated P. sativum Rubisco highlights the conservation of the form I Rubisco hexadecameric complex. The electron density places the substrate in the active site at the interface of the large-subunit dimers. Lys201 in the active site is not carbamylated as expected for this non-activated structure. Some heterogeneity in the small-subunit sequence is noted, as well as possible variations in the conformation and contacts of ribulose 1,5-bisphosphate in the large-subunit active sites. Overall, the active-site conformation most closely correlates with the ‘closed’ conformation observed in other substrate/inhibitor-bound Rubisco structures. PMID:23295478

  2. Dark-induced and organ-specific expression of two asparagine synthetase genes in Pisum sativum.

    PubMed Central

    Tsai, F Y; Coruzzi, G M

    1990-01-01

    Nucleotide sequence analysis of cDNAs for asparagine synthetase (AS) of Pisum sativum has uncovered two distinct AS mRNAs (AS1 and AS2) encoding polypeptides that are highly homologous to the human AS enzyme. The amino-terminal residues of both AS1 and AS2 polypeptides are identical to the glutamine-binding domain of the human AS enzyme, indicating that the full-length AS1 and AS2 cDNAs encode glutamine-dependent AS enzymes. Analysis of nuclear DNA shows that AS1 and AS2 are each encoded by single genes in P.sativum. Gene-specific Northern blot analysis reveals that dark treatment induces high-level accumulation of AS1 mRNA in leaves, while light treatment represses this effect as much as 30-fold. Moreover, the dark-induced accumulation of AS1 mRNA was shown to be a phytochrome-mediated response. Both AS1 and AS2 mRNAs also accumulate to high levels in cotyledons of germinating seedlings and in nitrogen-fixing root nodules. These patterns of AS gene expression correlate well with the physiological role of asparagine as a nitrogen transport amino acid during plant development. Images Fig. 4. Fig. 5. Fig. 6. Fig. 7. PMID:1968003

  3. Does low uranium concentration generates phytotoxic symptoms in Pisum sativum L. in nutrient medium?

    PubMed

    Tawussi, Frank; Walther, Clemens; Gupta, Dharmendra K

    2017-09-06

    Due to excessive mining and use of radionuclide especially uranium (U) and its fission products, numerous health hazards as well as environmental contamination worldwide have been created. The present study focused on demonstrating whether low concentration of U treatment in liquid nutient medium may translocate traces of U in plants and in fruits of Pisum sativum after 30 and 60 days of exposure for the safe use as a food supplement for human/animals. Hydroponically grown plants (in amended Hoagland medium) were treated with two different concentrations of uranium ([U] = 100 and 500 nM, respectively). Plants showed a decrease in total chlorophyll after 60 days of treatment. On the other hand, Eh of the nutrient medium was not affected from the initial days till 60 days of treatment, but pH of nutrient medium was increased upon durations, highest at 60 days of treatment. In seeds, micro/macro elements were under limit as well as U concentration was also under detection limit. We did not observe any U in the above ground parts (shoots/seeds) of the plant, i.e., under detection limit. Our observation suggests that P. sativum plants may be useful to grow at low radionuclide [U]-contaminated areas for safe human/animal use, but for other fission products, we have to investigate further for the safe human/animal use.

  4. Cloning, overexpression, purification and preliminary crystallographic studies of a mitochondrial type II peroxiredoxin from Pisum sativum

    SciTech Connect

    Barranco-Medina, Sergio; López-Jaramillo, Francisco Javier; Bernier-Villamor, Laura; Sevilla, Francisca; Lázaro, Juan-José

    2006-07-01

    The isolation, purification, crystallization and molecular-replacement solution of mitochondrial type II peroxiredoxin from P. sativum is reported. A cDNA encoding an open reading frame of 199 amino acids corresponding to a type II peroxiredoxin from Pisum sativum with its transit peptide was isolated by RT-PCR. The 171-amino-acid mature protein (estimated molecular weight 18.6 kDa) was cloned into the pET3d vector and overexpressed in Escherichia coli. The recombinant protein was purified and crystallized by the hanging-drop vapour-diffusion technique. A full data set (98.2% completeness) was collected using a rotating-anode generator to a resolution of 2.8 Å from a single crystal flash-cooled at 100 K. X-ray data revealed that the protein crystallizes in space group P1, with unit-cell parameters a = 61.88, b = 66.40, c = 77.23 Å, α = 102.90, β = 104.40, γ = 99.07°, and molecular replacement using a theoretical model predicted from the primary structure as a search model confirmed the presence of six molecules in the unit cell as expected from the Matthews coefficient. Refinement of the structure is in progress.

  5. Rates of sugar uptake by guard cell protoplasts of pisum sativum L. Related To the solute requirement for stomatal opening

    PubMed

    Ritte; Rosenfeld; Rohrig; Raschke

    1999-10-01

    We wished to determine whether the capacity of the sugar uptake mechanisms of guard cells of the Argenteum mutant of pea (Pisum sativum L.) sufficed to support a concurrent stomatal opening movement. Sugar uptake by guard cell protoplasts was determined by silicone-oil-filtering centrifugation. The protoplasts took up [(14)C]glucose, [(14)C]fructose, and [(14)C]sucrose (Suc), apparently in symport with protons. Mannose, galactose, and fructose competed with Glc for transport by a presumed hexose carrier. The uptake of Glc saturated with a K(m) of 0.12 mM and a V(max) of 19 fmol cell(-1) h(-1). At external concentrations <1 mM, the uptake of Suc was slower than that of Glc. It exhibited a saturating component with a K(m) varying between 0.25 and 0.8 mM and a V(max) between 1 and 10 fmol cell(-1) h(-1), and at external concentrations >1 mM, a non-saturating component. At apoplastic sugar concentrations below 4 mM, sugar import was estimated to be mainly in the form of hexoses and too slow to support a simultaneous stomatal opening movement. If, however, during times of high photosynthesis and transpiration, the apoplastic Suc concentration rose and entered the range of non-saturating import, absorbed Suc could replace potassium malate as the osmoticum for the maintenance of stomatal opening.

  6. Characterization of Low-Strigolactone Germplasm in Pea (Pisum sativum L.) Resistant to Crenate Broomrape (Orobanche crenata Forsk.).

    PubMed

    Pavan, Stefano; Schiavulli, Adalgisa; Marcotrigiano, Angelo Raffaele; Bardaro, Nicoletta; Bracuto, Valentina; Ricciardi, Francesca; Charnikhova, Tatsiana; Lotti, Concetta; Bouwmeester, Harro; Ricciardi, Luigi

    2016-10-01

    Crenate broomrape (Orobanche crenata Forsk.) is a devastating parasitic weed threatening the cultivation of legumes around the Mediterranean and in the Middle East. So far, only moderate levels of resistance were reported to occur in pea (Pisum sativum L.) natural germplasm, and most commercial cultivars are prone to severe infestation. Here, we describe the selection of a pea line highly resistant to O. crenata, following the screening of local genetic resources. Time series observations show that delayed emergence of the parasite is an important parameter associated with broomrape resistance. High performance liquid chromatography connected to tandem mass spectrometry analysis and in vitro broomrape germination bioassays suggest that the resistance mechanism might involve the reduced secretion of strigolactones, plant hormones exuded by roots and acting as signaling molecules for the germination of parasitic weeds. Two years of replicated trials in noninfested fields indicate that the resistance is devoid of pleiotropic effects on yield, in contrast to pea experimental mutants impaired in strigolactone biosynthesis and, thus, is suitable for use in breeding programs.

  7. Palladium uptake by Pisum sativum: partitioning and effects on growth and reproduction.

    PubMed

    Ronchini, Matteo; Cherchi, Laura; Cantamessa, Simone; Lanfranchi, Marco; Vianelli, Alberto; Gerola, Paolo; Berta, Graziella; Fumagalli, Alessandro

    2015-05-01

    Environmental palladium levels are increasing because of anthropogenic activities. The considerable mobility of the metal, due to solubilisation phenomena, and its known bioavailability may indicate interactions with higher organisms. The aim of the study was to determine the Pd uptake and distribution in the various organs of the higher plant Pisum sativum and the metal-induced effects on its growth and reproduction. P. sativum was grown in vermiculite with a modified Hoagland's solution of nutrients in the presence of Pd at concentrations ranging 0.10-25 mg/L. After 8-10 weeks in a controlled environment room, plants were harvested and dissected to isolate the roots, stems, leaves, pods and peas. The samples were analysed for Pd content using AAS and SEM-EDX. P. sativum absorbed Pd, supplied as K₂PdCl₄, beginning at seed germination and continuing throughout its life. Minimal doses (0.10-1.0 mg Pd/L) severely inhibited pea reproductive processes while showing a peculiar hormetic effect on root development. Pd concentrations ≥1 mg/L induced developmental delay, with late growth resumption, increased leaf biomass (up to 25%) and a 15-20% reduction of root mass. Unsuccessful repeated blossoming efforts led to misshapen pods and no seed production. Photosynthesis was also disrupted. The absorbed Pd (ca. 0.5 % of the supplied metal) was primarily fixed in the root, specifically in the cortex, reaching concentrations up to 200 μg/g. The metal moved through the stem (up to 1 μg/g) to the leaves (2 μg/g) and pods (0.3 μg/g). The presence of Pd in the pea fruits, together with established evidence of environmental Pd accumulation and bioavailability, suggests possible contamination of food plants and propagation in the food chain and must be the cause for concern.

  8. The CRC orthologue from Pisum sativum shows conserved functions in carpel morphogenesis and vascular development.

    PubMed

    Fourquin, Chloé; Primo, Amparo; Martínez-Fernández, Irene; Huet-Trujillo, Estefanía; Ferrándiz, Cristina

    2014-11-01

    CRABS CLAW (CRC) is a member of the YABBY family of transcription factors involved in carpel morphogenesis, floral determinacy and nectary specification in arabidopsis. CRC orthologues have been functionally characterized across angiosperms, revealing additional roles in leaf vascular development and carpel identity specification in Poaceae. These studies support an ancestral role of CRC orthologues in carpel development, while roles in vascular development and nectary specification appear to be derived. This study aimed to expand research on CRC functional conservation to the legume family in order to better understand the evolutionary history of CRC orthologues in angiosperms. CRC orthologues from Pisum sativum and Medicago truncatula were identified. RNA in situ hybridization experiments determined the corresponding expression patterns throughout flower development. The phenotypic effects of reduced CRC activity were investigated in P. sativum using virus-induced gene silencing. CRC orthologues from P. sativum and M. truncatula showed similar expression patterns, mainly restricted to carpels and nectaries. However, these expression patterns differed from those of other core eudicots, most importantly in a lack of abaxial expression in the carpel and in atypical expression associated with the medial vein of the ovary. CRC downregulation in pea caused defects in carpel fusion and style/stigma development, both typically associated with CRC function in eudicots, but also affected vascular development in the carpel. The data support the conserved roles of CRC orthologues in carpel fusion, style/stigma development and nectary development. In addition, an intriguing new aspect of CRC function in legumes was the unexpected role in vascular development, which could be shared by other species from widely diverged clades within the angiosperms, suggesting that this role could be ancestral rather than derived, as so far generally accepted. © The Author 2014. Published by

  9. The CRC orthologue from Pisum sativum shows conserved functions in carpel morphogenesis and vascular development

    PubMed Central

    Fourquin, Chloé; Primo, Amparo; Martínez-Fernández, Irene; Huet-Trujillo, Estefanía; Ferrándiz, Cristina

    2014-01-01

    Background and Aims CRABS CLAW (CRC) is a member of the YABBY family of transcription factors involved in carpel morphogenesis, floral determinacy and nectary specification in arabidopsis. CRC orthologues have been functionally characterized across angiosperms, revealing additional roles in leaf vascular development and carpel identity specification in Poaceae. These studies support an ancestral role of CRC orthologues in carpel development, while roles in vascular development and nectary specification appear to be derived. This study aimed to expand research on CRC functional conservation to the legume family in order to better understand the evolutionary history of CRC orthologues in angiosperms. Methods CRC orthologues from Pisum sativum and Medicago truncatula were identified. RNA in situ hybridization experiments determined the corresponding expression patterns throughout flower development. The phenotypic effects of reduced CRC activity were investigated in P. sativum using virus-induced gene silencing. Key Results CRC orthologues from P. sativum and M. truncatula showed similar expression patterns, mainly restricted to carpels and nectaries. However, these expression patterns differed from those of other core eudicots, most importantly in a lack of abaxial expression in the carpel and in atypical expression associated with the medial vein of the ovary. CRC downregulation in pea caused defects in carpel fusion and style/stigma development, both typically associated with CRC function in eudicots, but also affected vascular development in the carpel. Conclusions The data support the conserved roles of CRC orthologues in carpel fusion, style/stigma development and nectary development. In addition, an intriguing new aspect of CRC function in legumes was the unexpected role in vascular development, which could be shared by other species from widely diverged clades within the angiosperms, suggesting that this role could be ancestral rather than derived, as so far

  10. Primary and Secondary Abscission in Pisum sativum and Euphorbia pulcherrima-How Do They Compare and How Do They Differ?

    PubMed

    Hvoslef-Eide, Anne K; Munster, Cristel M; Mathiesen, Cecilie A; Ayeh, Kwadwo O; Melby, Tone I; Rasolomanana, Paoly; Lee, YeonKyeong

    2015-01-01

    Abscission is a highly regulated and coordinated developmental process in plants. It is important to understand the processes leading up to the event, in order to better control abscission in crop plants. This has the potential to reduce yield losses in the field and increase the ornamental value of flowers and potted plants. A reliable method of abscission induction in poinsettia (Euphorbia pulcherrima) flowers has been established to study the process in a comprehensive manner. By correctly decapitating buds of the third order, abscission can be induced in 1 week. AFLP differential display (DD) was used to search for genes regulating abscission. Through validation using qRT-PCR, more information of the genes involved during induced secondary abscission have been obtained. A study using two pea (Pisum sativum) mutants in the def (Developmental funiculus) gene, which was compared with wild type peas (tall and dwarf in both cases) was performed. The def mutant results in a deformed, abscission-less zone instead of normal primary abscission at the funiculus. RNA in situ hybridization studies using gene sequences from the poinsettia differential display, resulted in six genes differentially expressed for abscission specific genes in both poinsettia and pea. Two of these genes are associated with gene up- or down-regulation during the first 2 days after decapitation in poinsettia. Present and previous results in poinsettia (biochemically and gene expressions), enables a more detailed division of the secondary abscission phases in poinsettia than what has previously been described from primary abscission in Arabidopsis. This study compares the inducible secondary abscission in poinsettia and the non-abscising mutants/wild types in pea demonstrating primary abscission zones. The results may have wide implications on the understanding of abscission, since pea and poinsettia have been separated for 94-98 million years in evolution, hence any genes or processes in common

  11. Primary and Secondary Abscission in Pisum sativum and Euphorbia pulcherrima—How Do They Compare and How Do They Differ?

    PubMed Central

    Hvoslef-Eide, Anne K.; Munster, Cristel M.; Mathiesen, Cecilie A.; Ayeh, Kwadwo O.; Melby, Tone I.; Rasolomanana, Paoly; Lee, YeonKyeong

    2016-01-01

    Abscission is a highly regulated and coordinated developmental process in plants. It is important to understand the processes leading up to the event, in order to better control abscission in crop plants. This has the potential to reduce yield losses in the field and increase the ornamental value of flowers and potted plants. A reliable method of abscission induction in poinsettia (Euphorbia pulcherrima) flowers has been established to study the process in a comprehensive manner. By correctly decapitating buds of the third order, abscission can be induced in 1 week. AFLP differential display (DD) was used to search for genes regulating abscission. Through validation using qRT-PCR, more information of the genes involved during induced secondary abscission have been obtained. A study using two pea (Pisum sativum) mutants in the def (Developmental funiculus) gene, which was compared with wild type peas (tall and dwarf in both cases) was performed. The def mutant results in a deformed, abscission-less zone instead of normal primary abscission at the funiculus. RNA in situ hybridization studies using gene sequences from the poinsettia differential display, resulted in six genes differentially expressed for abscission specific genes in both poinsettia and pea. Two of these genes are associated with gene up- or down-regulation during the first 2 days after decapitation in poinsettia. Present and previous results in poinsettia (biochemically and gene expressions), enables a more detailed division of the secondary abscission phases in poinsettia than what has previously been described from primary abscission in Arabidopsis. This study compares the inducible secondary abscission in poinsettia and the non-abscising mutants/wild types in pea demonstrating primary abscission zones. The results may have wide implications on the understanding of abscission, since pea and poinsettia have been separated for 94–98 million years in evolution, hence any genes or processes in common

  12. Analysis of nodule senescence in pea (Pisum sativum L.) using laser microdissection, real-time PCR, and ACC immunolocalization.

    PubMed

    Serova, Tatiana A; Tikhonovich, Igor A; Tsyganov, Viktor E

    2017-05-01

    A delay in the senescence of symbiotic nodules could prolong active nitrogen fixation, resulting in improved crop yield and a reduced need for chemical fertilizers. The molecular genetic mechanisms underlying nodule senescence have not been extensively studied with a view to breeding varieties with delayed nodule senescence. In such studies, plant mutants with the phenotype of premature degradation of symbiotic structures are useful models to elucidate the genetic basis of nodule senescence. Using a dataset from transcriptome analysis of Medicago truncatula Gaertn. nodules and previous studies on pea (Pisum sativum L.) nodules, we developed a set of molecular markers based on genes that are known to be activated during nodule senescence. These genes encode cysteine proteases, a thiol protease, a bZIP transcription factor, enzymes involved in the biosynthesis of ethylene (ACS2 for ACC synthase and ACO1 for ACC oxidase) and ABA (AO3 for aldehyde oxidase), and an enzyme involved in catabolism of gibberellins (GA 2-oxidase). We analyzed the transcript levels of these genes in the nodules of two pea wild-types (cv. Sparkle and line Sprint-2) and two mutant lines, one showing premature nodule senescence (E135F (sym13)) and one showing no morphological signs of symbiotic structure degradation (Sprint-2Fix(-) (sym31)). Real-time PCR analyses revealed that all of the selected genes showed increased transcript levels during nodule aging in all phenotypes. Remarkably, at 4 weeks after inoculation (WAI), the transcript levels of all analyzed genes were significantly higher in the early senescent nodules of the mutant line E135F (sym13) and in nodules of the mutant Sprint-2Fix(-) (sym31) than in the active nitrogen-fixing nodules of wild-types. In contrast, the transcript levels of the same genes of both wild-types were significantly increased only at 6 WAI. We evaluated the expression of selected markers in the different histological nodule zones of pea cv. Sparkle and its

  13. Internode length in Pisum. Gene na may block gibberellin synthesis between ent-7. cap alpha. -hydroxykaurenoic acid and biggerellin A/sub 12/-aldehyde. [Pisum sativum

    SciTech Connect

    Ingram, T.J.; Reid, J.B.

    1987-04-01

    The elongation response of the gibberellin (GA) deficient genotypes na, ls, and lh of peas (Pisum sativum L.) to a range of GA-precursors was examined. Plants possessing gene na did not respond to precursors in the GA biosynthetic pathway prior to GA/sub 12/-aldehyde. In contrast, plants possessing lh and ls responded as well as wild-type plants (dwarfed with AMO-1618) to these compounds. The results suggest that GA biosynthesis is blocked prior to ent-kaurene in the lh and ls mutants and between ent-7..cap alpha..-hydroxykaurenoic acid and GA/sub 12/-aldehyde in the na mutant. Feeds of ent(/sup 3/H)kaurenoic acid and (/sup 2/H)GA/sub 12/-aldehyde to a range of genotypes supported the above conclusions. The na line WL1766 was shown by gas chromatography-mass spectrometry (GC-MS) to metabolize(/sup 2/H)GA/sub 12/-aldehyde to a number of (/sup 2/H)C/sub 19/-GAs including GA/sub 1/. However, there was no indication in na genotypes for the metabolism of ent-(/sup 3/H)kaurenoic acid to these GAs. In contrast, the expanding shoot tissue of all Na genotypes examined metabolized ent-(/sup 3/H)kaurenoic acid to radioactive compounds that co-chromatographed with GA/sub 1/, GA/sub 8/, GA/sub 20/, and GA/sub 29/. However, insufficient material was present for unequivocal identification of the metabolites. The radioactive profiles from HPLC of extracts of the node treated with ent-(/sup 3/H)kaurenoic acid were similar for both Na and na plants and contained ent-16..cap alpha..,17-dihydroxykaurenoic acid and ent-6..cap alpha..,7..cap alpha..,16..beta..,17-tetrahydroxykaurenoic acid (both characterized by GC-MS), suggesting that the metabolites arose from side branches of the main GA-biosynthetic pathway. Thus, both Na and na plants appear capable of ent-7..cap alpha..-hydroxylation.

  14. Auxin effects on in vitro and in vivo protein phosphorylation in pea. [Pisum sativum

    SciTech Connect

    Gallagher, S.R.; Ray, P.M.

    1987-04-01

    Terminal 8mm sections from the third internode of dark grown 7 day old Pisum sativum cv Alaska seedlings were separated into membrane and soluble fractions. SDS gradient PAGE identified approximately 50 in vivo phosphorylated proteins and proved superior to 2-D SDS PAGE in terms of resolution and repeatability. Addition of indoleacetic acid (IAA), fusicoccin, or 2,4 dichlorophenoxyacetic acid to membranes resulted in no detectable change in the number or phosphorylation level of the labeled proteins during in vitro phosphorylation in the presence of submicromolar concentrations of calcium. Similar results were obtained with soluble proteins. In the absence of calcium, the level of in vitro protein phosphorylation was much less, but not auxin effects could be identified. Furthermore, treatment of the sections with IAA in vivo followed by cell fractionation and in vitro phosphorylation failed to identify auxin responsive proteins. Lastly, when sections were labeled with /sup 32/P inorganic phosphate in the presence of 17 uM IAA, no auxin specific changes were found in the level of phosphorylation or in the number of phosphorylated proteins. Auxin effects on phosphorylation are thus slight or below their detection limit.

  15. [Biochemical changes associated with cadmium and copper stress in germinating pea seeds (Pisum sativum L.)].

    PubMed

    Mihoub, Asma; Chaoui, Abdelilah; El Ferjani, Ezzedine

    2005-01-01

    Seeds of pea (Pisum sativum L.) were germinated for four days over two sheets of filter paper moistened with H2O (control) and 5 mM Cd(NO3)2 or CuSO4 (treated). The relationship between heavy-metal stress and breakdown of storage compounds was studied. Germination rate and growth of radicle decreased, while the water content in stressed seeds remained around the control values. Cotyledons changed their biochemical constituents: disorders in the contents of micronutrients (Fe, Mn, Zn), free amino acids and soluble sugars were found. Decline of alpha-amylase activity as well as acid phosphatase were also observed, whereas beta-amylase and alkaline phosphatase ones were not modified by heavy-metal treatments. These results suggest that the inhibition of seed germinations after exposure to cadmium or copper is not the consequence of starvation in water uptake by seed tissues, but may be due to a failure in the reserve mobilization process from cotyledons.

  16. A proteomic approach to decipher chilling response from cold acclimation in pea (Pisum sativum L.).

    PubMed

    Dumont, Estelle; Bahrman, Nasser; Goulas, Estelle; Valot, Benoît; Sellier, Hélène; Hilbert, Jean-Louis; Vuylsteker, Christophe; Lejeune-Hénaut, Isabelle; Delbreil, Bruno

    2011-01-01

    Two pea lines (Pisum sativum L.) with contrasted behaviours towards chilling and subsequent frost were studied by a proteomic approach to better understand cold acclimation. Following a chilling period, the Champagne line becomes tolerant to frost whereas Terese remains sensitive. Variance analysis allowed to select 260 statistically variable spots with 68 identified proteins (35 in leaves, 18 in stems, and 15 in roots). These proteins were shared out in proteins related to chilling response or cold acclimation. The better adaptation of Champagne to chilling might be related to a higher content in proteins involved in photosynthesis and in defence mechanisms. Moreover Champagne might prevent freezing damage particularly thanks to a higher constitutive expression of housekeeping proteins related to Terese. After three days of subsequent frost, proteomes of previously chilled plants also showed significant differences compared to unchilled plants. Out of 112 statistically variable spots (44 in leaves, 38 in stems, and 30 in roots), 32 proteins were identified. These proteins were related to frost response or frost resistance. It seems that Champagne could resist to frost with the reorientation of the energy metabolism. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

  17. Agronomical factors influencing the legumin/vicilin ratio in pea (Pisum sativum L.) seeds.

    PubMed

    Mertens, Cécile; Dehon, Lydie; Bourgeois, Audrey; Verhaeghe-Cartrysse, Christine; Blecker, Christophe

    2012-06-01

    Many research studies have investigated the impact of agronomical factors on the protein content of pea (Pisum sativum). This study aimed to establish if a correlation exists between protein content and legumin/vicilin (L/V) ratio in pea seeds and to identify agronomical factors that have an impact on this ratio. The L/V ratio was positively correlated with protein content (r = 0.58, P ≤ 0.01), but no linear regression was applicable (adjusted R(2) = 0.31). Both variety and cultivation year had a highly significant effect on the ratio (P < 0.001). The interaction between these two factors was also highly significant (P < 0.001), some varieties being less sensitive to climatic conditions than others. Cultivation location had a highly significant effect (P < 0.01). There was no interaction between variety and location. Seeding density had a highly significant effect on the ratio (P < 0.01), with a saturation effect above 60 seeds m(-2). Further studies should establish if a linear regression model can be applied to pea varieties independently. Varieties with a stable L/V ratio can prove to be useful in the food industry. Other agronomical factors (soil type and seeding density) should be considered for the production of pea seeds with a specific L/V ratio. Copyright © 2011 Society of Chemical Industry.

  18. Profile and Functional Properties of Seed Proteins from Six Pea (Pisum sativum) Genotypes

    PubMed Central

    Barac, Miroljub; Cabrilo, Slavica; Pesic, Mirjana; Stanojevic, Sladjana; Zilic, Sladjana; Macej, Ognjen; Ristic, Nikola

    2010-01-01

    Extractability, extractable protein compositions, technological-functional properties of pea (Pisum sativum) proteins from six genotypes grown in Serbia were investigated. Also, the relationship between these characteristics was presented. Investigated genotypes showed significant differences in storage protein content, composition and extractability. The ratio of vicilin:legumin concentrations, as well as the ratio of vicilin + convicilin: Legumin concentrations were positively correlated with extractability. Our data suggest that the higher level of vicilin and/or a lower level of legumin have a positive influence on protein extractability. The emulsion activity index (EAI) was strongly and positively correlated with the solubility, while no significant correlation was found between emulsion stability (ESI) and solubility, nor between foaming properties and solubility. No association was evident between ESI and EAI. A moderate positive correlation between emulsion stability and foam capacity was observed. Proteins from the investigated genotypes expressed significantly different emulsifying properties and foam capacity at different pH values, whereas low foam stability was detected. It appears that genotype has considerable influence on content, composition and technological-functional properties of pea bean proteins. This fact can be very useful for food scientists in efforts to improve the quality of peas and pea protein products. PMID:21614186

  19. Hydrogen sulfide alleviates hypoxia-induced root tip death in Pisum sativum.

    PubMed

    Cheng, Wei; Zhang, Liang; Jiao, Chengjin; Su, Miao; Yang, Tao; Zhou, Lina; Peng, Renyi; Wang, Ranran; Wang, Chongying

    2013-09-01

    Flooding of soils often results in hypoxic conditions surrounding plant roots, which is a harmful abiotic stress to crops. Hydrogen sulfide (H2S) is a highly diffusible, gaseous molecule that modulates cell signaling and is involved in hypoxia signaling in animal cells. However, there have been no previous studies of H2S in plant cells in response to hypoxia. The effects of H2S on hypoxia-induced root tip death were studied in pea (Pisum sativum) via analysis of endogenous H2S and reactive oxygen species (ROS) levels. The activities of key enzymes involved in antioxidative and H2S metabolic pathways were determined using spectrophotometric assays. Ethylene was measured by gas chromatography. We found that exogenous H2S pretreatment dramatically alleviated hypoxia-induced root tip death by protecting root tip cell membranes from ROS damage induced by hypoxia and by stimulating a quiescence strategy through inhibiting ethylene production. Conversely, root tip death induced by hypoxia was strongly enhanced by inhibition of the key enzymes responsible for endogenous H2S biosynthesis. Our results demonstrated that exogenous H2S pretreatment significantly alleviates hypoxia-induced root tip death in pea seedlings and, therefore, enhances the tolerance of the plant to hypoxic stress. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  20. Transfer cell wall ingrowths and transport capacity in pea leaf discs. [Pisum sativum cv

    SciTech Connect

    Wimmers, L.E.; Turgeon, R.

    1986-04-01

    Transfer cell wall ingrowths are thought to increase transport capacity by increasing plasmelemma surface area. Little direct evidence for this hypothesis exists since experimental systems in which the surface area of wall ingrowths can be modulated have not been available. They grew Pisum sativum cv. Little Marvel plants under three light regimes (150, 500, 1000 umol photons m/sup -2/ sec/sup -1/) using 1000 watt Sylvania Metal Halide lamps. Wall ingrowths in minor vein phloem parenchyma cells were analyzed morphometrically from electron micrographs and a positive correlation was found between light intensity and extent of wall ingrowths. Vein loading was assayed by floating abraded leaf discs on /sup 14/C-sucrose (1 mM). There was a positive correlation between uptake and transfer cell wall surface area, although the latter increased more than the former. No significant differences were found in vein length, numbers of phloem elements, or phloem cross sectional areas. Changes in light intensity after a leaf reached maturity did not change uptake potential over a period of at least three days.

  1. Transfer cell wall ingrowths and vein loading characteristics in pea leaf discs. [Pisum sativum

    SciTech Connect

    Wimmers, L.E.; Turgeon, R.

    1987-04-01

    Transfer cell wall ingrowths are thought to increase transport capacity by increasing plasmalemma surface area. Leaf minor vein phloem transfer cells presumably enhance phloem loading. In Pisum sativum cv. Little marvel grown under different light regimes (150 to 1000 ..mu..mol photons m/sup -2/ sec/sup -1/) there is a positive correlation between light intensity and wall ingrowth area in phloem transfer cells. The extent of ingrowth and correlation to light intensity is greatest in minor veins, decreasing as vein size increases. Vein loading was assayed by floating abraded leaf discs on /sup 14/C-sucrose (10 mM). There is a positive correlation between uptake and transfer cell wall area, although the latter increased more than the former. The difference in uptake is stable throughout the photoperiod, and is also stable in mature leaves for at least four days after plants are transfered to a different light intensity. Sucrose uptake is biphasic. The saturable component of uptake is sensitive to light intensity, the Km for sucrose is negatively correlated to light intensity, while V/sub max/remains unchanged.

  2. NADH induces the generation of superoxide radicals in leaf peroxisomes. [Pisum sativum L

    SciTech Connect

    del Rio, L.A.; Sandalio, L.M.; Palma, J.M. ); Fernandez, V.M.; Ruperez, F.L. )

    1989-03-01

    In peroxisomes isolated from pea leaves (Pisum sativum L.) the production of superoxide free radicals (O{sub 2}{sup {minus}}) by xanthine and NADH was investigated. In peroxisomal membranes, 100 micromolar NADH induced the production of O{sub 2}{sup {minus}} radicals. In the soluble fractions of peroxisomes, no generation of O{sub 2}{sup {minus}} radicals was observed by incubation with either NADH or xanthine, although xanthine oxidase was found located predominantly in the matrix of peroxisomes. The failure of xanthine to induce superoxide generation was probably due to the inability to fully suppress the endogenous Mn-superoxide dismutase activity by inhibitors which were inactive against xanthine oxidase. The generation of superoxide radicals in leaf peroxisomes together with the recently described production of these oxygen radicals in glyoxysomes suggests that O{sub 2}{sup {minus}} generation could be a common metabolic property of peroxisomes and further supports the existence of active oxygen-related roles for peroxisomes in cellular metabolism.

  3. Nitrite reduction and carbohydrate metabolism in plastids purified from roots of Pisum sativum L.

    PubMed

    Bowsher, C G; Hucklesby, D P; Emes, M J

    1989-03-01

    Intact preparations of plastids from pea (Pisum sativum L.) roots have been used to investigate the metabolism of glucose-6-phosphate and reduction of inorganic nitrite within these organelles. The ability of hexose-phosphates to support nitrite reduction was dependent on the integrity of the preparation and was barely measurable in broken organelles. In intact plastids, nitrite was reduced most effectively in the presence of glucose-6-phosphate (Glc6P), fructose-6-phosphate and ribose-5-phosphate and to a lesser extent glucose-1-phosphate. The Km (Glc6P) of plastid-located Glc6P dehydrogenase (EC 1.1.1.49) and Glc6P-dependent nitrite reduction were virtually identical (0.68 and 0.66 mM respectively) and a similar relationship was observed between fructose-6-phosphate, hexose-phosphate isomerase (EC 5.3.1.9) and nitrite reduction. The pattern of release of CO2 from different carbon atoms of Glc6P supplied to root plastids, indicates the operation of both glycolysis and the oxidative pentose-phosphate pathway with some recycling in the latter. During nitrite reduction the evolution of CO2 from carbon atom 1 of Glc6P was stimulated but not from carbon atoms 2, 3, 4, or 6. The importance of these results with regard to the regulation of the pathways of carbohydrate oxidation and nitrogen assimilation within root plastids is discussed.

  4. Induction of Root Nodule Senescence by Combined Nitrogen in Pisum sativum L 1

    PubMed Central

    Chen, Pin-Ching; Phillips, Donald A.

    1977-01-01

    Root nodule senescence induced by nitrate and ammonium in Pisum sativum L. was defined by determining nitrogenase activity and leghemoglobin content with the acetylene reduction and pyridine hemochrome assays. Root systems supplied with 100 mm KNO3 or 100 mm NH4Cl exhibited a decrease in nitrogenase activity followed by a decline in leghemoglobin content. Increasing the CO2 concentration from 0.000320 atm to 0.00120 atm had no effect on the time course of root nodule senescence when 20 mm KNO3 was supplied to the roots; in vitro nitrate reductase activity was detected in leaves and roots, but not bacteroids. Nitrate appeared in leaves, roots, and the nodule cytosol fraction but not bacteroids when 20 mm KNO3 was supplied to roots. When nitrate entered through the shoots, however, no root nodule senescence was observed, and no nitrate was detected in root or nodule cytosol fractions although nitrate and nitrate reductase were found in leaves. The results suggest that nitrate does not induce root nodule senescence through competition between nitrate reductase and nitrogenase for products of photosynthesis. PMID:16659869

  5. Nitrogen Stress and Apparent Photosynthesis in Symbiotically Grown Pisum sativum L. 1

    PubMed Central

    Dejong, Ted M.; Phillips, Donald A.

    1981-01-01

    Pea plants (Pisum sativum L. cv. Alaska) were inoculated individually with one of 15 Rhizobium leguminosarum strains and grown under uniform environmental conditions in the absence of combined N. Differences in effectiveness of the Rhizobium strains produced plants with differing rates of whole plant apparent N2 fixation and total N content at the same morphological stage of development. Plants were analyzed to determine interactions between N2 fixation, N allocation, apparent photosynthesis, and growth. Total leaf N increased linearly with total N2 fixation (R2 = 0.994). The proportion of total N allocated to leaves, the per cent N content of individual leaves, and the photosynthetic efficiency of individual leaves showed a curvilinear response with increasing plant N content. Differences in allocation patterns of leaf N between plants with low and high N content resulted in differences in the relationship between total N content and plant dry weight. Results from this study show that N2 fixation interacts with leaf photosynthetic efficiency and plant growth in a manner that is dependent upon the allocation of symbiotically fixed N. PMID:16661907

  6. Effects of SO2 on Stomatal Metabolism in Pisum sativum L. 1

    PubMed Central

    Rao, I. Madhusudana; Amundson, Robert G.; Alscher-Herman, Ruth; Anderson, Louise E.

    1983-01-01

    Pea (Pisum sativum L. cv `Little Marvel') plants were exposed to SO2 for short term (3 hours) and long term (2 days) at 0.2 and at 0.5 microliter per liter (ppm) levels. The effect of this treatment on the activity of phosphoenolpyruvate carboxylase, NAD- and NADP-malate dehydrogenases, and alanine aminotransferase from epidermis and whole leaves was investigated. Short-term exposure to SO2 at 0.2 or 0.5 ppm decreased the activity of the carboxylase and the dehydrogenases in the epidermis. In contrast, the activity of the same three enzymes increased in whole leaves with either short- or long-term exposure to SO2. Alanine aminotransferase in epidermis or whole leaves was not much affected by short-term exposure, but the epidermal activity was decreased and whole leaf activity was increased with long-term exposure. SO2 exposure which was initiated prior to illumination decreased the free thiol content of both epidermis and of whole leaf. Net photosynthesis was reversibly inhibited by long-term exposure to SO2 at 0.5 ppm. No effect of 0.5 ppm SO2 on stomatal conductance was detectable after 3 hours. Stomatal conductance appeared to decrease after longer exposure times (2 days) at 0.5 ppm. PMID:16663045

  7. Rapidly Induced Wound Ethylene from Excised Segments of Etiolated Pisum sativum L., cv. Alaska

    PubMed Central

    Saltveit, Mikal E.; Dilley, David R.

    1978-01-01

    A rapidly induced, transitory increase in the rate of ethylene synthesis occurred in wounded tissue excised from actively growing regions of etiolated barley, cucumber, maize, oat, pea, tomato, and wheat seedlings. Cutting intact stems or excising 9-mm segments of tissue from near the apex of 7-day-old etiolated Pisum sativum L., cv. Alaska seedlings induced a remarkably consistent pattern of ethylene production. At 25 C, wound-induced ethylene production by segments excised 9 mm below the apical hook increased linearly after a lag of 26 minutes from 2.7 nanoliters per g per hour to the first maxium of 11.3 nanoliters per g per hour at 56 minutes. The rate of production then decreased to a minimum at 90 minutes, increased to a lower second maximum at 131 minutes, and subsequently declined over a period of about 100 minutes to about 4 nanoliters per g per hour. Removal of endogenous ethylene, before the wound response commenced, had no effect on the kinetics of ethylene production. Tissue containing large amounts of dissolved ethylene released it as an exponential decay with no lag period. Rapidly induced wound ethylene is synthesized by the tissue and is not merely the result of facilitated diffusion of ethylene already present in the tissue through the newly exposed cut surfaces. Previously wounded apical sections did not exhibit a second response when rewounded. No significant correlation was found between wound-induced ethylene synthesis and either CO2 or ethane production. PMID:16660312

  8. Phytochrome regulation of gibberellin metabolism in shoots of dwarf Pisum sativum L

    SciTech Connect

    Campell, B.R.

    1986-01-01

    To study the effect of light on the recessive dwarfing allele, le, of Pisum sativum L., etiolated, paclobutrazol treated LeLe (cv. Alaska) and lele (cv. Progress) pea seedlings were transferred to different light regimes. The growth response of both genotypes to applications of GA/sub 1/, GA/sub 20/, and steviol was measured over 48 hours using position transducers. Both genotypes responded to GA/sub 1/ under red irradiation and in darkness. The LeLe plants grew in response to steviol and GA/sub 20/ under red irradiation and in darkness. The lele plants responded to steviol and GA/sub 20/ in darkness, but showed a much smaller response when red irradiated. The red effect on lele plants was reversible by far-red irradiation. (/sup 3/H)GA/sub 20/ was applied to nana pea seedlings, homozygous for le, grown under different light regimes. Radioactive metabolites were later extracted from the shoots of the treated plants. Both the free acid and conjugate pools were analyzed by reversed phase HPLC, and some radioactive metabolites were tentatively identified by comparing their retention times to those of authentic (/sup 3/H)GAs.

  9. Pre-fractionation strategies to resolve pea (Pisum sativum) sub-proteomes

    PubMed Central

    Meisrimler, Claudia-Nicole; Menckhoff, Ljiljana; Kukavica, Biljana M.; Lüthje, Sabine

    2015-01-01

    Legumes are important crop plants and pea (Pisum sativum L.) has been investigated as a model with respect to several physiological aspects. The sequencing of the pea genome has not been completed. Therefore, proteomic approaches are currently limited. Nevertheless, the increasing numbers of available EST-databases as well as the high homology of the pea and medicago genome (Medicago truncatula Gaertner) allow the successful identification of proteins. Due to the un-sequenced pea genome, pre-fractionation approaches have been used in pea proteomic surveys in the past. Aside from a number of selective proteome studies on crude extracts and the chloroplast, few studies have targeted other components such as the pea secretome, an important sub-proteome of interest due to its role in abiotic and biotic stress processes. The secretome itself can be further divided into different sub-proteomes (plasma membrane, apoplast, cell wall proteins). Cell fractionation in combination with different gel-electrophoresis, chromatography methods and protein identification by mass spectrometry are important partners to gain insight into pea sub-proteomes, post-translational modifications and protein functions. Overall, pea proteomics needs to link numerous existing physiological and biochemical data to gain further insight into adaptation processes, which play important roles in field applications. Future developments and directions in pea proteomics are discussed. PMID:26539198

  10. Biological changes of green pea (Pisum sativum L.) by selenium enrichment.

    PubMed

    Garousi, Farzaneh; Kovács, Béla; Domokos-Szabolcsy, Éva; Veres, Szilvia

    2017-03-01

    Supplement of common fertilizers with selenium (Se) for crop production will be an effective way to produce selenium-rich food and feed. The value of green pea seeds and forages as alternative protein source can be improved by using agronomic biofortification. Therefore, biological changes of green pea (Pisum sativum L.) and influences of inorganic forms of Se (sodium selenite and sodium selenate) at different concentrations on the accumulation of magnesium (Mg) and phosphorus (P) were investigated in greenhouse experiment. 3 mg kg(-1) of selenite had positive effects to enhance photosynthetic attributes and decrease lipid peroxidation significantly. At the same time, Se accumulation increased in all parts of plant by increasing Se supply. Moreover, Mg and P accumulations were significantly increased at 3 mg kg(-1) selenite and 1 mg kg(-1) selenate treatments, respectively. By contrast higher selenite concentrations (≥30 mg kg(-1)) exerted toxic effects on plants. Relative chlorophyll content, actual photochemical efficiency of PSII (ФPSII) and Mg accumulation showed significant decrease while membrane lipid peroxidation increased. Thus, the present findings prove Se biofortification has positive effects on biological traits of green pea to provide it as a proper functional product.

  11. Application of SNPs to improve yield of Pisum sativum L. (pea).

    PubMed

    Mehmood, Ansar; Murtaza, Ghulam

    2017-06-01

    Nanotechnology opens an enormous scope of novel application in the fields of biotechnology and agricultural industries, because nanoparticles (NPs) have unique physicochemical properties, i.e. high surface area, high reactivity, tunable pore size and particle morphology. Present study was carried out to determine the role of silver NPs (SNPs) to improve yield of Pisum sativum L. SNPs (10-100 nm) were synthesised by green method using extract of Berberis lycium Royle. Pea seeds were soaked and seedling were foliage sprayed by 0, 30, 60 and 90 ppm SNPs. The experiment was arranged as split-split plot randomised complete block design with three replicates. The application of SNPs enhanced significantly number of seeds pod(-1), number of pods plant(-1), hundred seed weight, biological yield and green pod yield over control. The highest yield was found when 60 ppm SNPs were applied. However, exposure to 90 ppm SNPs, the yield of the pea decreased significantly as compared with 30 and 60 ppm. This research shows that SNPs have definite ability to improve growth and yield of crops. Nevertheless, a comprehensive experimentation is needed to establish the most appropriate concentration, size and mode of application of SNPs for higher growth and maximum yield of pea.

  12. Rapid wall relaxation in elongating tissues. [Glycine max (L. ); Pisum sativum L

    SciTech Connect

    Matyssek, R.; Maruyama, S.; Boyer, J.S. )

    1988-01-01

    Reported differences in the relaxation of cell walls in enlarging stem tissues of soybean (Glycine max (L.) Merr.) and pea (Pisum sativum L.) cause measurements of the yield threshold turgor, an important growth parameter, to be in doubt. Using the pressure probe and guillotine psychrometer, the authors investigated wall relaxation in these species by excising the elongating tissue in air to remove the water supply. The authors found that the rapid kinetics usually exhibited by soybean could be delayed and made similar to the slow kinetics previously reported for pea if slowly growing or mature tissue was left attached to the rapidly growing tissue when relaxation was initiated. The greater the amount of attached tissue, the slower the relaxation, suggesting that slowly growing tissue acted as a water source. Consistent with this concept was a lower water potential in the rapidly elongating tissue than in the slowly growing tissue. If this tissue was removed from pea, relaxation became as rapid as usually exhibited by soybean. It is concluded that the true relaxation of cell walls to the yield threshold requires only a few minutes and that the yield threshold in the intact plant before excision. Under these conditions, the yield threshold was close to the turgor in the intact plant regardless of the species.

  13. Purification and Characterization of a Lectin from Green Split Peas (Pisum sativum).

    PubMed

    Ng, Tzi Bun; Chan, Yau Sang; Ng, Charlene Cheuk Wing; Wong, Jack Ho

    2015-11-01

    Lectins have captured the attention of a large number of researchers on account of their various exploitable activities, including antitumor, immunomodulatory, antifungal, as well as HIV reverse transcriptase inhibitory activities. A mannose/glucose-specific lectin was isolated from green split peas (a variety of Pisum sativum) and characterized. The purification step involved anion-exchange chromatography on a DEAE-cellulose column, cation-exchange chromatography on an SP-Sepharose column, and gel filtration by fast protein liquid chromatography (FPLC) on Superdex 200. The purified lectin had a native molecular mass of around 50 kDa as determined by size exclusion chromatography. It appeared as a heterotetramer, composed of two distinct polypeptide bands with a molecular mass of 6 and 19 kDa, respectively, in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The N-terminal sequence of green split pea lectin shows some degree of homology compared to lectins from other legume species. Its hemagglutinating activity was inhibited by glucose, mannose, and sucrose, and attenuated at pH values higher than 12 or lower than 3. Hemagglutinating activity was preserved at temperatures lower than 80 °C. The lectin did not show antifungal activity toward fungi including Fusarium oxysporum, Botrytis cinerea, and Mycosphaerella arachidicola. Green split pea lectin showed a mitogenic effect toward murine splenocytes and could inhibit the activity of HIV-1 reverse transcriptase.

  14. Control of storage-protein synthesis during seed development in pea (Pisum sativum L.).

    PubMed

    Gatehouse, J A; Evans, I M; Bown, D; Croy, R R; Boulter, D

    1982-10-15

    The tissue-specific syntheses of seed storage proteins in the cotyledons of developing pea (Pisum sativum L.) seeds have been demonstrated by estimates of their qualitative and quantitative accumulation by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and rocket immunoelectrophoresis respectively. Vicilin-fraction proteins initially accumulated faster than legumin, but whereas legumin was accumulated throughout development, different components of the vicilin fraction had their predominant periods of synthesis at different stages of development. The translation products in vitro of polysomes isolated from cotyledons at different stages of development reflected the synthesis in vivo of storage-protein polypeptides at corresponding times. The levels of storage-protein mRNA species during development were estimated by 'Northern' hybridization using cloned complementary-DNA probes. This technique showed that the levels of legumin and vicilin (47000-Mr precursors) mRNA species increased and decreased in agreement with estimated rates of synthesis of the respective polypeptides. The relative amounts of these messages, estimated by kinetic hybridization were also consistent. Legumin mRNA was present in leaf poly(A)+ RNA at less than one-thousandth of the level in cotyledon poly(A)+ (polyadenylated) RNA, demonstrating tissue-specific expression. Evidence is presented that storage-protein mRNA species are relatively long-lived, and it is suggested that storage-protein synthesis is regulated primarily at the transcriptional level.

  15. Influence of ammonium chloride on the nitrogenase activity of nodulated pea plants (Pisum sativum).

    PubMed Central

    Houwaard, F

    1978-01-01

    A study was made on the short-term effect of ammonium ions on the nitrogenase activity of pea root nodules. Nodulated pea plants (Pisum sativum), having reached maximum acetylene-reducing activity, were supplied with NH4Cl (20 mM). Nitrogenase activity of intact plants, detached nodules, and isolated bacteroids was measured at differed time intervals. A significant drop (20 to 40%) in the acetylene-reducing activity of treated intact plants and their detached nodules was observed after 1 day. No drop in the nitrogenase activity of bacteroids (assayed aerobically, or anaerobically after treatment with ethylenediaminetetraacetic acid-toluene) occurred for 2 to 4 days after the addition of NH4+ to the plants, depending on cultural conditions. From these results it is concluded that the adverse effect of NH4+ on acetylene reduction by intact plants and detached nodules during the first 2 days is not due to a decrease in the amount of nitrogenase in the bacteroids. It is suggested that the effect has to be attributed to a reduced supply to the bacteroids of energy-delivery photosynthates. PMID:677873

  16. Conserved thioredoxin fold is present in Pisum sativum L. sieve element occlusion-1 protein.

    PubMed

    Tuteja, Narendra; Umate, Pavan; Tuteja, Renu

    2010-06-01

    Homology-based three-dimensional model for Pisum sativum sieve element occlusion 1 (Ps.SEO1) (forisomes) protein was constructed. A stretch of amino acids (residues 320 to 456) which is well conserved in all known members of forisomes proteins was used to model the 3D structure of Ps.SEO1. The structural prediction was done using Protein Homology/analogY Recognition Engine (PHYRE) web server. Based on studies of local sequence alignment, the thioredoxin-fold containing protein [Structural Classification of Proteins (SCOP) code d1o73a_], a member of the glutathione peroxidase family was selected as a template for modeling the spatial structure of Ps.SEO1. Selection was based on comparison of primary sequence, higher match quality and alignment accuracy. Motif 1 (EVF) is conserved in Ps.SEO1, Vicia faba (Vf.For1) and Medicago truncatula (Mt.SEO3); motif 2 (KKED) is well conserved across all forisomes proteins and motif 3 (IGYIGNP) is conserved in Ps.SEO1 and Vf.For1.

  17. Purification and characterization of ornithine transcarbamylase from pea (Pisum sativum L.)

    NASA Technical Reports Server (NTRS)

    Slocum, R. D.; Richardson, D. P.

    1991-01-01

    Pea (Pisum sativum) ornithine transcarbamylase (OTC) was purified to homogeneity from leaf homogenates in a single-step procedure, using delta-N-(phosphonacetyl)-L-ornithine-Sepharose 6B affinity chromatography. The 1581-fold purified OTC enzyme exhibited a specific activity of 139 micromoles citrulline per minute per milligram of protein at 37 degrees C, pH 8.5. Pea OTC represents approximately 0.05% of the total soluble protein in the leaf. The molecular weight of the native enzyme was approximately 108,200, as estimated by Sephacryl S-200 gel filtration chromatography. The purified protein ran as a single molecular weight band of 36,500 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results suggest that the pea OTC is a trimer of identical subunits. The overall amino acid composition of pea OTC is similar to that found in other eukaryotic and prokaryotic OTCs, but the number of arginine residues is approximately twofold higher. The increased number of arginine residues probably accounts for the observed isoelectric point of 7.6 for the pea enzyme, which is considerably more basic than isoelectric point values that have been reported for other OTCs.

  18. Digestion of fibre polysaccharides of pea (Pisum sativum) hulls, carrot and cabbage by adult cockerels.

    PubMed

    Longstaff, M; McNab, J M

    1989-11-01

    Characterization of the carbohydrates of pea (Pisum sativum) hulls, carrot and cabbage using both colorimetric and gas-liquid chromatographic techniques permitted a detailed investigation into the extent of digestion of differing types of fibre. These digestion studies were greatly aided by the development of a rapid bioassay employing starved adult cockerels. Total collection of undigested residues, uncontaminated by food spillage, could be made from trays placed under the cockerels. Chemical analysis showed that pea hulls consisted mainly of fibre with very little available carbohydrate present, whereas more than half of freeze-dried carrot and cabbage consisted of available carbohydrate (sucrose, glucose, fructose, starch) and consequently considerably less fibre was present. The fibre of carrot and cabbage was similarly composed of nearly equal amounts of neutral and acidic polysaccharides, whereas pea-hull fibre had four times as much neutral as acidic polysaccharides. The digestibility of total neutral polysaccharides from all three foodstuffs was extremely low. However, there appeared to be preferential digestion of polysaccharides composed of rhamnose, arabinose and galactose residues, all associated with pectic material, in contrast to the indigestibility of polysaccharides composed of fucose, xylose and glucose. Acidic polysaccharides were digested to a greater extent than neutral ones, and those of carrot and cabbage more so than pea hulls. The polysaccharides which were the most soluble were also the most digestible, but due to the arbitrariness of polysaccharide solubility, quantification of their total digestibility per se was considered not possible.

  19. Pre-fractionation strategies to resolve pea (Pisum sativum) sub-proteomes.

    PubMed

    Meisrimler, Claudia-Nicole; Menckhoff, Ljiljana; Kukavica, Biljana M; Lüthje, Sabine

    2015-01-01

    Legumes are important crop plants and pea (Pisum sativum L.) has been investigated as a model with respect to several physiological aspects. The sequencing of the pea genome has not been completed. Therefore, proteomic approaches are currently limited. Nevertheless, the increasing numbers of available EST-databases as well as the high homology of the pea and medicago genome (Medicago truncatula Gaertner) allow the successful identification of proteins. Due to the un-sequenced pea genome, pre-fractionation approaches have been used in pea proteomic surveys in the past. Aside from a number of selective proteome studies on crude extracts and the chloroplast, few studies have targeted other components such as the pea secretome, an important sub-proteome of interest due to its role in abiotic and biotic stress processes. The secretome itself can be further divided into different sub-proteomes (plasma membrane, apoplast, cell wall proteins). Cell fractionation in combination with different gel-electrophoresis, chromatography methods and protein identification by mass spectrometry are important partners to gain insight into pea sub-proteomes, post-translational modifications and protein functions. Overall, pea proteomics needs to link numerous existing physiological and biochemical data to gain further insight into adaptation processes, which play important roles in field applications. Future developments and directions in pea proteomics are discussed.

  20. Development and characterization of 37 novel EST-SSR markers in Pisum sativum (Fabaceae)1

    PubMed Central

    Zhuang, Xiaofeng; McPhee, Kevin E.; Coram, Tristan E.; Peever, Tobin L.; Chilvers, Martin I.

    2013-01-01

    • Premise of the study: Simple sequence repeat markers were developed based on expressed sequence tags (EST-SSR) and screened for polymorphism among 23 Pisum sativum individuals to assist development and refinement of pea linkage maps. In particular, the SSR markers were developed to assist in mapping of white mold disease resistance quantitative trait loci. • Methods and Results: Primer pairs were designed for 46 SSRs identified in EST contiguous sequences assembled from a 454 pyrosequenced transcriptome of the pea cultivar, ‘LIFTER’. Thirty-seven SSR markers amplified PCR products, of which 11 (30%) SSR markers produced polymorphism in 23 individuals, including parents of recombinant inbred lines, with two to four alleles. The observed and expected heterozygosities ranged from 0 to 0.43 and from 0.31 to 0.83, respectively. • Conclusions: These EST-SSR markers for pea will be useful for refinement of pea linkage maps, and will likely be useful for comparative mapping of pea and as tools for marker-based pea breeding. PMID:25202482

  1. Effects of SO/sub 2/ on stomatal metabolism in Pisum sativum L

    SciTech Connect

    Rao, I.M.; Amundson, R.G.; Alshcer-Herman, R.; Anderson, L.E.

    1983-01-01

    Pea (Pisum sativum L. cv 'Little Marvel') plants were exposed to SO/sub 2/ for short term (3 hours) and long term (2 days) at 0.2 and at 0.5 microliter per liter (ppm) levels. The effect of this treatment on the activity of phosphoenolpyruvate carboxylase, NAD- and NADP-malate dehydrogenases, and alanine aminotransferase from epidermis and whole leaves was investigated. Short-term exposure to SO/sub 2/ at 0.2 or 0.5 ppm decreased the activity of the carboxylase and the dehydrogenases in the epidermis. In contrast, the activity of the same three enzymes increased in whole leaves with either short- or long-term exposure to SO/sub 2/. Alanine aminotransferase in epidermis or whole leaves was not much affected by short-term exposure, but the epidermal activity was decreased and whole leaf activity was increased with long-term exposure. SO/sub 2/ exposure which was initiated prior to illumination decreased the free thiol content of both epidermis and of whole leaf. Net photosynthesis was reversibly inhibited by long-term exposure to SO/sub 2/ at 0.5 ppm. No effect of 0.5 ppm SO/sub 2/ on stomatal conductance was detectable after 3 hours. Stomatal conductance appeared to decrease after longer exposure times (2 days) at 0.5 ppm.

  2. Comparative effect of calcium and EDTA on arsenic uptake and physiological attributes of Pisum sativum.

    PubMed

    Rafiq, Marina; Shahid, Muhammad; Abbas, Ghulam; Shamshad, Saliha; Khalid, Sana; Niazi, Nabeel Khan; Dumat, Camille

    2017-07-03

    In this study, we determined the effect of ethylenediaminetetraacetic acid (EDTA) and calcium (Ca) on arsenic (As) uptake and toxicity to Pisum sativum. Plants were treated with three levels of As (25, 125, and 250 µM) in the presence and absence of three levels of Ca (1, 5, and 10 mM) and EDTA (25, 125, and 250 µM). Exposure to As caused an overproduction of hydrogen peroxide (H2O2) in roots and leaves, which induced lipid peroxidation and decreased pigment contents. Application of both Ca and EDTA significantly reduced As accumulation by pea, Ca being more effective in reducing As accumulation. Both Ca and EDTA enhanced As-induced H2O2 production, but reduced lipid peroxidation. In the case of pigment contents, EDTA significantly reduced pigment contents, whereas Ca significantly enhanced pigment contents compared to As alone. The effect of As treatment in the presence and absence of EDTA and Ca was more pronounced in younger leaves compared to older leaves. The effect of amendments varied greatly with their applied levels, as well as type and age of plant organs. Importantly, due to possible precipitation of Ca-As compounds, the soils with higher levels of Ca ions are likely to be less prone to food chain contamination.

  3. Study of effect of AC and DC magnetic fields on growth of Pisum sativum seeds

    NASA Astrophysics Data System (ADS)

    Bahar, Mahmood; Yasaie Mehrjardi, Yasaman; Sojoodi, Jaleh; Bayani, Hosien; Kazem Salem, Mohammad

    2013-08-01

    This paper concentrates on the effect of the AC and DC magnetic fields on plant growth. The effect of AC magnetic field with intensities of 2.25, 1.66 and 1.49 mT and DC magnetic field with intensities of 3.6, 2.41 and 2.05 mT in exposure durations of 2, 4, 6, 8, 10 and 12 min on two groups of dry and wet Pisum sativum seedlings was studied. In each experiment 10 seeds were used; the experiments were repeated three times for each group and there was a sham exposed group for comparison purposes. The light cycle was 12 h light/12 h darkness and the temperature was 25 ± 1° C. The index of growth is considered to be the root and stem elongation on the sixth day. It was observed that AC magnetic field has a positive effect on the growth in all durations and intensities. Moreover, it is highlighted that during the experiments, the mean growth of dry seedlings significantly increased by a factor of 11 in AC magnetic field with the lowest intensity of 1.49 mT (p < 0.05). It was also shown that AC magnetic fields had a more positive effect on the growth of plants in comparison to DC magnetic fields.

  4. Study of DC and AC electric field effect on Pisum sativum seeds growth

    NASA Astrophysics Data System (ADS)

    Mahmood, Bahar; Jaleh, Sojoodi; Yasaman, Yasaie

    2014-07-01

    In this research the effect of electric field on two groups of wet and dry Pisum sativum seeds growth was studied. To generate the required electric field a parallel-plate capacitor with round copper plates of 30 cm diameter was used. The experiments were performed once in fixed exposure duration of 8 min in variable DC electric field of 0.25-1.5 kV/m. The other experiments were performed in variable fields of 50-125 kV/m in fixed exposure duration of 8 min, in two groups of AC and DC electric fields. The experiments were repeated three times. In each experiment 10 seeds were used and there was a sham exposed group for comparison, too. After application of electric field, the seeds were kept for six days in the same growth chamber with the temperature of 25 ± 1 °C and 12 h light/12 h darkness. On the 6th day length of stems and height of roots were measured. After doing statistical analysis, in low intensities of DC electric field, the highest significant increase of mean growth (The average of stem length and the height of roots) was seen in 1.5 kV/m in wet seeds. In high intensities of DC and AC electric fields, the highest significant increase of mean growth was seen in AC electric field of 100 kV/m in wet seeds.

  5. Alkane biosynthesis by decarbonylation of aldehydes catalyzed by a particulate preparation from Pisum sativum

    SciTech Connect

    Cheesbourgh, T.M.; Kolattukudy, P.E.

    1984-11-01

    Mechanism of enzymatic conversion of a fatty acid to the corresponding alkane by the loss of the carboxyl carbon was investigated with particulate preparations from Pisum sativum. A heavy particulate preparation (sp. gr., 1.30 g/cm/sup 3/) isolated by two density-gradient centrifugation steps catalyzed conversion of octadecanal to heptadecane and CO. Experiments with (1-/sup 3/H, 1-/sup 14/C)octadecanal showed the stoichiometry of the reaction and retention of the aldehydic hydrogen in the alkane during this enzymatic decarbonylation. This decarbonylase showed an optimal pH of 7.0 and a K/sub m/ of 35 ..mu..M for the aldehyde. This enzyme was severly inhibited by metal ion chelators and showed no requirement for any cofactors. Microsomal preparations and the particulate fractions from the first density-gradient step catalyzed acyl-CoA reduction to the corresponding aldehyde. Electron microscopic examination showed the presence of fragments of cell wall/cuticle but no vesicles in the decarbonylase preparation. It is concluded that the aldehydes produced by the acyl-CoA reductase located in the endomembranes of the epidermal cells are converted to alkanes by the decarbonylase located in the cell wall/cuticle region. 20 references, 4 figures, 1 tables.

  6. Dark Respiration Protects Photosynthesis Against Photoinhibition in Mesophyll Protoplasts of Pea (Pisum sativum) 1

    PubMed Central

    Saradadevi, Kanakagiri; Raghavendra, Agepati S.

    1992-01-01

    The optimal light intensity required for photosynthesis by mesophyll protoplasts of pea (Pisum sativum) is about 1250 microeinsteins per square meter per second. On exposure to supra-optimal light intensity (2500 microeinsteins per square meter per second) for 10 min, the protoplasts lost 30 to 40% of their photosynthetic capacity. Illumination with normal light intensity (1250 microeinsteins per square meter per second) for 10 min enhanced the rate of dark respiration in protoplasts. On the other hand, when protoplasts were exposed to photoinhibitory light, their dark respiration also was markedly reduced along with photosynthesis. The extent of photoinhibition was increased when protoplasts were incubated with even low concentrations of classic respiratory inhibitors: 1 micromolar antimycin A, 1 micromolar sodium azide, and 1 microgram per milliliter oligomycin. At these concentrations, the test inhibitors had very little or no effect directly on the process of photosynthetic oxygen evolution. The promotion of photoinhibition by inhibitors of oxidative electron transport (antimycin A, sodium azide) and phosphorylation (oligomycin) was much more pronounced than that by inhibitors of glycolysis and tricarboxylic acid cycle (sodium fluoride and sodium malonate, respectively). We suggest that the oxidative electron transport and phosphorylation in mitochondria play an important role in protecting the protoplasts against photoinhibition of photosynthesis. Our results also demonstrate that protoplasts offer an additional experimental system for studies on photoinhibition. PMID:16668993

  7. Studies on antioxidative enzymes induced by cadmium in pea plants (Pisum sativum).

    PubMed

    Pandey, Nalini; Singh, Gaurav Kumar

    2012-03-01

    Pea plants (Pisum sativum cv. Swati) exposed to different concentration of cadmium (50,100, 200 microM Cd) under controlled glass house conditions were quantified for different physiological parameters and antioxidative enzymes. In pea plants, Cd produced a significant inhibition of growth and induced chlorosis, marginal yellowing and necrosis in young leaves, the effect being most pronounced at 200 microM Cd supply. An alteration in the activated oxygen metabolism of pea plants were also detected as evidenced by an increase in concentration of H2O2 and TBARS along with decrease in the chlorophyll and carotenoid concentration in leaves. Cadmium toxicity induced an increase in non-protein thiol, ascorbate, proline and cysteine concentration. A significant increment in the activity of SOD, APX and GR, and a decrease in CAT was observed as a result of Cd treatment. The enhanced activity of SOD and inhibition of CAT and POD produces a high build up of H2O2 which appears to be the main cause of oxidative stress due to Cd toxicity in pea plants.

  8. Micromonospora halotolerans sp. nov., isolated from the rhizosphere of a Pisum sativum plant.

    PubMed

    Carro, Lorena; Pukall, Rüdiger; Spröer, Cathrin; Kroppenstedt, Reiner M; Trujillo, Martha E

    2013-06-01

    A filamentous actinomycete strain designated CR18(T) was isolated on humic acid agar from the rhizosphere of a Pisum sativum plant collected in Spain. This isolate was observed to grow optimally at 28 °C, pH 7.0 and in the presence of 5 % NaCl. Phylogenetic analyses based on the 16S rRNA gene sequence indicated a close relationship with the type strains of Micromonospora chersina and Micromonospora endolithica. A further analysis based on a concatenated DNA sequence stretch of 4,523 bp that included partial sequences of the atpD, gyrB, recA, rpoB and 16S rRNA genes clearly differentiated the new strain from recognized Micromonospora species compared. DNA-DNA hybridization studies further supported the taxonomic position of strain CR18(T) as a novel genomic species. Chemotaxonomic analyses which included whole cell sugars, polar lipids, fatty acid profiles and menaquinone composition confirmed the affiliation of the new strain to the genus Micromonospora and also highlighted differences at the species level. These studies were finally complemented with an array of physiological tests to help differentiate between the new strain and its phylogenetic neighbours. Consequently, strain CR18(T) (= CECT 7890(T) = DSM 45598(T)) is proposed as the type strain of a novel species, Micromonospora halotolerans sp. nov.

  9. PsPMEP, a pollen-specific pectin methylesterase of pea (Pisum sativum L.).

    PubMed

    Gómez, María Dolores; Renau-Morata, Begoña; Roque, Edelín; Polaina, Julio; Beltrán, José Pío; Cañas, Luis A

    2013-09-01

    Pectin methylesterases (PMEs) are a family of enzymes involved in plant reproductive processes such as pollen development and pollen tube growth. We have isolated and characterized PsPMEP, a pea (Pisum sativum L.) pollen-specific gene that encodes a protein with homology to PMEs. Sequence analysis showed that PsPMEP belongs to group 2 PMEs, which are characterized by the presence of a processable amino-terminal PME inhibitor domain followed by the catalytic PME domain. Moreover, PsPMEP contains several motifs highly conserved among PMEs with the essential amino acid residues involved in enzyme substrate binding and catalysis. Northern blot and in situ hybridization analyses showed that PsPMEP is expressed in pollen grains from 4 days before anthesis till anther dehiscence and in pollinated carpels. In the PsPMEP promoter region, we have identified several conserved cis-regulatory elements that have been associated with gene pollen-specific expression. Expression analysis of PsPMEP promoter fused to the uidA reporter gene in Arabidopsis thaliana plants showed a similar expression pattern when compared with pea, indicating that this promoter is also functional in a non-leguminous plant. GUS expression was detected in mature pollen grains, during pollen germination, during pollen tube elongation along the transmitting tract, and when the pollen tube reaches the embryo sac in the ovule.

  10. Purification and characterization of ornithine transcarbamylase from pea (Pisum sativum L.)

    NASA Technical Reports Server (NTRS)

    Slocum, R. D.; Richardson, D. P.

    1991-01-01

    Pea (Pisum sativum) ornithine transcarbamylase (OTC) was purified to homogeneity from leaf homogenates in a single-step procedure, using delta-N-(phosphonacetyl)-L-ornithine-Sepharose 6B affinity chromatography. The 1581-fold purified OTC enzyme exhibited a specific activity of 139 micromoles citrulline per minute per milligram of protein at 37 degrees C, pH 8.5. Pea OTC represents approximately 0.05% of the total soluble protein in the leaf. The molecular weight of the native enzyme was approximately 108,200, as estimated by Sephacryl S-200 gel filtration chromatography. The purified protein ran as a single molecular weight band of 36,500 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results suggest that the pea OTC is a trimer of identical subunits. The overall amino acid composition of pea OTC is similar to that found in other eukaryotic and prokaryotic OTCs, but the number of arginine residues is approximately twofold higher. The increased number of arginine residues probably accounts for the observed isoelectric point of 7.6 for the pea enzyme, which is considerably more basic than isoelectric point values that have been reported for other OTCs.

  11. Organic meat quality of dual purpose young bulls supplemented with pea (Pisum sativum L.) or soybean.

    PubMed

    Corazzin, Mirco; Piasentier, Edi; Saccà, Elena; Bazzoli, Ilario; Bovolenta, Stefano

    2017-07-12

    One of the main constraints established by organic legislation that limits the development of the rearing of young bulls is the ban on the use of genetically modified organisms (GMO). Most of the worldwide cultivated soybean is GMO, therefore the use of alternative protein sources should be evaluated. In this study, the effect of dietary substitution of soybean with pea (Pisum sativum L.) on carcass characteristics and meat quality of dual purpose young bulls reared following the organic method was investigated. Twenty-four young bulls of Rendena breed were randomly assigned to two diet treatments differing in protein supplement (soybean (SB) or field pea (FP)). Carcass characteristics and meat chemical composition, colour, cooking loss and Warner-Bratzler shear force did not differ between groups. Regarding meat fatty acid composition, SB showed higher concentrations of C18:0 and C18:1 t and lower C16:1n-9c, C14:0, C17:1n-9c and C18:1n-9c than FP. In descriptive sensory analysis, trained judges were not able to differentiate meats from SB and FP, which also had similar overall liking expressed by consumers. The results of this study indicate that FP can replace SB in the diet of dual purpose young bulls with only a minor influence on fatty acid composition and no effect on carcass characteristics and meat quality. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  12. Cloning, overexpression, purification and preliminary crystallographic studies of a mitochondrial type II peroxiredoxin from Pisum sativum

    PubMed Central

    Barranco-Medina, Sergio; López-Jaramillo, Francisco Javier; Bernier-Villamor, Laura; Sevilla, Francisca; Lázaro, Juan-José

    2006-01-01

    A cDNA encoding an open reading frame of 199 amino acids corresponding to a type II peroxiredoxin from Pisum sativum with its transit peptide was isolated by RT-PCR. The 171-amino-acid mature protein (estimated molecular weight 18.6 kDa) was cloned into the pET3d vector and overexpressed in Escherichia coli. The recombinant protein was purified and crystallized by the hanging-drop vapour-diffusion technique. A full data set (98.2% completeness) was collected using a rotating-anode generator to a resolution of 2.8 Å from a single crystal flash-cooled at 100 K. X-ray data revealed that the protein crystallizes in space group P1, with unit-cell parameters a = 61.88, b = 66.40, c = 77.23 Å, α = 102.90, β = 104.40, γ = 99.07°, and molecular replacement using a theoretical model predicted from the primary structure as a search model confirmed the presence of six molecules in the unit cell as expected from the Matthews coefficient. Refinement of the structure is in progress. PMID:16820697

  13. Extracellular production of reactive oxygen species during seed germination and early seedling growth in Pisum sativum.

    PubMed

    Kranner, Ilse; Roach, Thomas; Beckett, Richard P; Whitaker, Claire; Minibayeva, Farida V

    2010-07-01

    Extracellularly produced reactive oxygen species (ROS) play key roles in plant development, but their significance for seed germination and seedling establishment is poorly understood. Here we report on the characteristics of extracellular ROS production during seed germination and early seedling development in Pisum sativum. Extracellular superoxide (O2(.-)) and hydrogen peroxide (H2O2) production and the activity of extracellular peroxidases (ECPOX) were determined spectrophotometrically, and O2(.-) was identified by electron paramagnetic resonance. Cell wall fractionation of cotyledons, seed coats and radicles was used in conjunction with polyacrylamide gel electrophoresis to investigate substrate specificity and molecular masses of O2(.-)-producing enzymes, and the forces that bind them to the cell wall. Seed imbibition was accompanied by an immediate, transient burst of redox activity that involved O2(.-) and other substances capable of oxidizing epinephrine, and also H2O2. At the final stages of germination, coinciding with radicle elongation, a second increase in O2(.-) but not H2O2 production occurred and was correlated with an increase in extracellular ECPOX activity. Electrophoretic analyses of cell wall fractions demonstrated the presence of enzymes capable of O2(.-) production. The significance of extracellular ROS production during seed germination and early seedling development, and also during seed aging, is discussed.

  14. Isozymes of beta-N-Acetylhexosaminidase from Pea Seeds (Pisum sativum L.).

    PubMed

    Harley, S M; Beevers, L

    1987-12-01

    Four isozymes of beta-N-acetylhexosaminidase (beta-NAHA) from pea seeds (Pisum sativum L.) have been separated, with one, designated beta-NAHA-II, purified to apparent homogeneity by means of an affinity column constructed by ligating p-aminophenyl-N-acetyl-beta-d-thioglucosaminide to Affi-Gel 202. The other three isozymes have been separated and purified 500- to 1750-fold by chromatography on Concanavalin A-Sepharose, Zn(2+) charged immobilized metal affinity chromatography, hydrophobic chromatography, and ion exchange chromatography on CM-Sephadex. All four isozymes are located in the protein bodies of the cotyledons. The molecular weight of each isozyme is 210,000. beta-NAHA-II is composed of two heterogenous subunits. The subunits are not held together by disulfide bonds, but sulfhydryl groups are important for catalysis. All four isozymes release p-nitrophenol from both p-nitrophenyl-N-acetyl-beta-d-glucosaminide and p-nitrophenyl-N-acetyl-beta-d-galactosaminide. The ratio of activity for hydrolysis of the two substrates is pH dependent. The K(m) value for the two substrates and pH optima of the isozymes are comparable to beta-NAHAs from other plant sources.

  15. Genetic control and identification of QTLs associated with visual quality traits of field pea (Pisum sativum L.).

    PubMed

    Ubayasena, Lasantha; Bett, Kirstin; Tar'an, Bunyamin; Warkentin, Thomas

    2011-04-01

    Visual quality of field pea (Pisum sativum L.) is one of the most important determinants of the market value of the harvested crop. Seed coat color, seed shape, and seed dimpling are the major components of visual seed quality of field pea and are considered as important breeding objectives. The objectives of this research were to study the genetics and to identify quantitative trait loci (QTLs) associated with seed coat color, seed shape, and seed dimpling of green and yellow field peas. Two recombinant inbred line populations (RILs) consisting of 120 and 90 lines of F(5)-derived F(7) (F(5:7)) yellow pea (P. sativum 'Alfetta' × P. sativum 'CDC Bronco') and green pea (P. sativum 'Orb' × P. sativum 'CDC Striker'), respectively, were evaluated over two years at two locations in Saskatchewan, Canada. Quantitative inheritance with polygenic control and transgressive segregation were observed for all visual quality traits studied. All 90 RILs of the green pea population and 92 selected RILs from the yellow pea population were screened using AFLP and SSR markers and two linkage maps were developed. Nine QTLs controlling yellow seed lightness, 3 for yellow seed greenness, 15 for seed shape, and 9 for seed dimpling were detected. Among them, five QTLs located on LG II, LG IV, and LG VII were consistent in at least two environments. The QTLs and their associated markers will be useful tools to assist pea breeding programs attempting to pyramid positive alleles for the traits.

  16. Wild peas vary in their cross-compatibility with cultivated pea (Pisum sativum subsp. sativum L.) depending on alleles of a nuclear-cytoplasmic incompatibility locus.

    PubMed

    Bogdanova, V S; Kosterin, O E; Yadrikhinskiy, A K

    2014-05-01

    Divergent wild and endemic peas differ in hybrid sterility in reciprocal crosses with cultivated pea depending on alleles of a nuclear 'speciation gene' involved in nuclear-cytoplasmic compatibility. In hybrids between cultivated and wild peas, nuclear-cytoplasmic conflict frequently occurs. One of the nuclear genes involved, Scs1, was earlier mapped on Linkage Group III. In reciprocal crosses of seven divergent pea accessions with cultivated P. sativum, some alleles of Scs1 manifested incompatibility with an alien cytoplasm as a decrease in pollen fertility to about 50 % in the heterozygotes and lack of some genotypic classes among F2 segregants. Earlier, we defined monophyletic evolutionary lineages A, B, C and D of pea according to allelic state of three markers, from nuclear, plastid and mitochondrial genomes. All tested representatives of wild peas from the lineages A and C exhibited incompatibility due to Scs1 deleterious effects in crosses with testerlines of P. sativum subsp. sativum (the common cultivated pea) at least in one direction. A wild pea from the lineage B and a cultivated pea from the lineage D were compatible with the testerline in both directions. The tested accession of cultivated P. abyssinicum (lineage A) was partially compatible in both directions. The Scs1 alleles of some pea accessions even originating from the same geographic area were remarkably different in their compatibility with cultivated Pisum sativum cytoplasm. Variability of a gene involved in reproductive isolation is of important evolutionary role and nominate Scs1 as a speciation gene.

  17. Transcriptomic analysis of Rhizobium leguminosarum biovar viciae in symbiosis with host plants Pisum sativum and Vicia cracca.

    PubMed

    Karunakaran, R; Ramachandran, V K; Seaman, J C; East, A K; Mouhsine, B; Mauchline, T H; Prell, J; Skeffington, A; Poole, P S

    2009-06-01

    Rhizobium leguminosarum bv. viciae forms nitrogen-fixing nodules on several legumes, including pea (Pisum sativum) and vetch (Vicia cracca), and has been widely used as a model to study nodule biochemistry. To understand the complex biochemical and developmental changes undergone by R. leguminosarum bv. viciae during bacteroid development, microarray experiments were first performed with cultured bacteria grown on a variety of carbon substrates (glucose, pyruvate, succinate, inositol, acetate, and acetoacetate) and then compared to bacteroids. Bacteroid metabolism is essentially that of dicarboxylate-grown cells (i.e., induction of dicarboxylate transport, gluconeogenesis and alanine synthesis, and repression of sugar utilization). The decarboxylating arm of the tricarboxylic acid cycle is highly induced, as is gamma-aminobutyrate metabolism, particularly in bacteroids from early (7-day) nodules. To investigate bacteroid development, gene expression in bacteroids was analyzed at 7, 15, and 21 days postinoculation of peas. This revealed that bacterial rRNA isolated from pea, but not vetch, is extensively processed in mature bacteroids. In early development (7 days), there were large changes in the expression of regulators, exported and cell surface molecules, multidrug exporters, and heat and cold shock proteins. fix genes were induced early but continued to increase in mature bacteroids, while nif genes were induced strongly in older bacteroids. Mutation of 37 genes that were strongly upregulated in mature bacteroids revealed that none were essential for nitrogen fixation. However, screening of 3,072 mini-Tn5 mutants on peas revealed previously uncharacterized genes essential for nitrogen fixation. These encoded a potential magnesium transporter, an AAA domain protein, and proteins involved in cytochrome synthesis.

  18. Transcriptomic Analysis of Rhizobium leguminosarum Biovar viciae in Symbiosis with Host Plants Pisum sativum and Vicia cracca▿ †

    PubMed Central

    Karunakaran, R.; Ramachandran, V. K.; Seaman, J. C.; East, A. K.; Mouhsine, B.; Mauchline, T. H.; Prell, J.; Skeffington, A.; Poole, P. S.

    2009-01-01

    Rhizobium leguminosarum bv. viciae forms nitrogen-fixing nodules on several legumes, including pea (Pisum sativum) and vetch (Vicia cracca), and has been widely used as a model to study nodule biochemistry. To understand the complex biochemical and developmental changes undergone by R. leguminosarum bv. viciae during bacteroid development, microarray experiments were first performed with cultured bacteria grown on a variety of carbon substrates (glucose, pyruvate, succinate, inositol, acetate, and acetoacetate) and then compared to bacteroids. Bacteroid metabolism is essentially that of dicarboxylate-grown cells (i.e., induction of dicarboxylate transport, gluconeogenesis and alanine synthesis, and repression of sugar utilization). The decarboxylating arm of the tricarboxylic acid cycle is highly induced, as is γ-aminobutyrate metabolism, particularly in bacteroids from early (7-day) nodules. To investigate bacteroid development, gene expression in bacteroids was analyzed at 7, 15, and 21 days postinoculation of peas. This revealed that bacterial rRNA isolated from pea, but not vetch, is extensively processed in mature bacteroids. In early development (7 days), there were large changes in the expression of regulators, exported and cell surface molecules, multidrug exporters, and heat and cold shock proteins. fix genes were induced early but continued to increase in mature bacteroids, while nif genes were induced strongly in older bacteroids. Mutation of 37 genes that were strongly upregulated in mature bacteroids revealed that none were essential for nitrogen fixation. However, screening of 3,072 mini-Tn5 mutants on peas revealed previously uncharacterized genes essential for nitrogen fixation. These encoded a potential magnesium transporter, an AAA domain protein, and proteins involved in cytochrome synthesis. PMID:19376875

  19. [Morphological Changes in the Structure of Blastozones during Fasciation of Pisum sativum L. and Arabidopsis thaliana (L.) Heynh].

    PubMed

    Bykova, E A; Labunskaya, E A; Chub, V V

    2015-01-01

    Different effects of fasciation on the structure of blastozones and ultimately on the morphology of the plants of the two families investigated was detected. The results of this study demonstrate that in Arabidopsis thaliana phyllotaxis changes, leading to the formation of whorled leaf aestivation (two or four leaf in the whorl). We also demonstrated that in Pisum sativum the number of folioles of a compound leaf increased, the number of stipules also increased, and stipules can grow together and form a structure similar to the sheath of Gramineae. Our results demonstrated that a common manifestation of fasciations in Arabidopsis and Pisum is the formation of a flattened meristem, which extended along the same line (axes of fasciation).

  20. Pea (Pisum sativum) cells arrested in G2 have nascent DNA with breaks between replicons and replication clusters

    SciTech Connect

    Van't Hof, J.

    1980-01-01

    DNA fiber autoradiography and alkaline sucrose sedimentation of DNA of cultured pea-root cells (Pisum sativum) arrested in G2 by carbohydrate starvation demonstrated that nascent DNA molecules of replicon (16 to 27 x 10/sup 6/D) and apparent cluster (approx. 330 x 10/sup 6/D) size were not joined. That the arrested cells were in G2 was confirmed by single-cell autoradiography and cytophotometry. In pea there are about 18 replicons per average cluster, 4.2 x 10/sup 3/ clusters, and 7.7 x 10/sup 4/ replicons per genome.

  1. Analysis of a diverse global Pisum sp. collection and comparison to a Chinese local P. sativum collection with microsatellite markers.

    PubMed

    Zong, Xuxiao; Redden, Robert J; Liu, Qingchang; Wang, Shumin; Guan, Jianping; Liu, Jin; Xu, Yanhong; Liu, Xiuju; Gu, Jing; Yan, Long; Ades, Peter; Ford, Rebecca

    2009-01-01

    Twenty-one informative microsatellite loci were used to assess and compare the genetic diversity among Pisum genotypes sourced from within and outside China. The Chinese germplasm comprised 1243 P. sativum genotypes from 28 provinces and this was compared to 774 P. sativum genotypes that represented a globally diverse germplasm collection, as well as 103 genotypes from related Pisum species. The Chinese P. sativum germplasm was found to contain genotypes genetically distinct from the global gene pool sourced outside China. The Chinese spring type genotypes were separate from the global gene pool and from the other main Chinese gene pool of winter types. The distinct Chinese spring gene pool comprised genotypes from Inner Mongolia and Sha'anxi provinces, with those from Sha'anxi showing the greatest diversity. The other main gene pool within China included both spring types from other northern provinces and winter types from central and southern China, plus some accessions from Inner Mongolia and Sha'anxi. A core collection of Chinese landraces chosen to represent molecular diversity was compared both to the wider Chinese collection and to a geographically diverse core collection of Chinese landraces. The average gene diversity and allelic richness per locus of both the micro-satellite based core and the wider collection were similar, and greater than the geographically diverse core. The genetic diversity of P. sativum within China appears to be quite different to that detected in the global gene pool, including the presence of several rare alleles, and may be a useful source of allelic variation for both major gene and quantitative traits.

  2. Purification, characterization and physiological role of sucrose synthase in the pea seed coat (Pisum sativum L.).

    PubMed

    Déjardin, A; Rochat, C; Maugenest, S; Boutin, J P

    1997-01-01

    The seed coat is a maternal organ which surrounds the embryo and is involved in the control of its nutrition. This study with pea (Pisum sativum L.) was conducted to understand more fully the sucrose/starch interconversions occurring in the seed coat. The concentrations of soluble sugars, the starch content, and the activities of the sucrose-metabolizing enzymes, sucrose synthase (Sus; EC 2.4.1.13), alkaline and soluble acid invertase (EC 3.2.1.26) and sucrose-phosphate synthase (SPS; EC 2.4.1.14) were compared at four developmental stages during seed filling. Among the four enzymes, only Sus activity was very high and strongly correlated with the starch concentration in the seed coat. Sucrose synthase catalyses the cleavage of sucrose in the presence of UDP into UDP-glucose and fructose. Sucrose synthase was purified from pea seed coats in a three-step protocol, consisting of diethylaminoethyl-Sephacel chromatography, gel filtration and affinity chromatography. The enzyme was characterized at the biochemical and molecular levels. Sucrose synthase exhibits biochemical properties which allow it to function in the direction of both sucrose cleavage and synthesis. The mass-action ratio of its four substrate was close to the theoretical equilibrium constant at the four developmental stages we studied. A labelling experiment on seed coats has shown that Sus activity is reversible in vivo and can produce 37% of neo-synthesized sucrose in the seed coat cells (minimum value). It is concluded that Sus could play a central role in the control of sucrose concentration in the seed coat cells in response to the demand for sucrose in the embryo during the development of the seed.

  3. Sitona lineatus (Coleoptera: Curculionidae) Larval Feeding on Pisum sativum L. Affects Soil and Plant Nitrogen.

    PubMed

    Cárcamo, Héctor A; Herle, Carolyn E; Lupwayi, Newton Z

    2015-01-01

    Adults of Sitona lineatus (pea leaf weevil, PLW) feed on foliage of several Fabaceae species but larvae prefer to feed on nodules of Pisum sativum L. and Vicia faba L. Indirectly, through their feeding on rhizobia, weevils can reduce soil and plant available nitrogen (N). However, initial soil N can reduce nodulation and damage by the weevil and reduce control requirements. Understanding these interactions is necessary to make integrated pest management recommendations for PLW. We conducted a greenhouse study to quantify nodulation, soil and plant N content, and nodule damage by weevil larvae in relation to soil N amendment with urea, thiamethoxam insecticide seed coating and crop stage. PLWs reduced the number of older tumescent (multilobed) nodules and thiamethoxam addition increased them regardless of other factors. Nitrogen amendment significantly increased soil available N (>99% nitrate) as expected and PLW presence was associated with significantly lower levels of soil N. PLW decreased plant N content at early flower and thiamethoxam increased it, particularly at late flower. The study illustrated the complexity of interactions that determine insect herbivory effects on plant and soil nutrition for invertebrates that feed on N-fixing root nodules. We conclude that effects of PLW on nodulation and subsequent effects on plant nitrogen are more pronounced during the early growth stages of the plant. This suggests the importance of timing of PLW infestation and may explain the lack of yield depression in relation to this pest observed in many field studies. Also, pea crops in soils with high levels of soil N are unlikely to be affected by this herbivore and should not require insecticide inputs. © Her Majesty the Queen in Right of Canada, as represented by the Minister of Agriculture and Agri-Food Canada, 2015.

  4. Variety discrimination in pea (Pisum sativum L.) by molecular, biochemical and morphological markers.

    PubMed

    Smykal, Peter; Horacek, Jiri; Dostalova, Radmila; Hybl, Miroslav

    2008-01-01

    The distinctness, uniformity and stability (DUS) requirements involve expensive, space- and time-consuming measurements of morphological traits. Moreover, for a majority of traits, interactions between genotype and environment complicate the evaluation. Molecular markers have a potential to facilitate this procedure, increase the reliability of decisions, and substantially save the time and space needed for experiments. We chose 25 varieties of pea (Pisum sativum L.) from the list of recommended varieties for cultivation in the Czech Republic, and made both a standard classification by 12 morphological descriptors and a classification by biochemical-molecular markers. Two isozyme systems, 10 microsatellite loci, 2 retrotransposons for multilocus inter-retrotransposon amplified polymorphism (IRAP), and 12 retrotransposon-based insertion polymorphism (RBIP) DNA markers were analysed. The main objective of the study was to examine the potential of each method for discrimination between pea varieties. The results demonstrate a high potential and resolving power of DNA-based methods. Superior in terms of high information content and discrimination power were SSR markers, owing to high allelic variation, which was the only biochemical-molecular method allowing clear identification of all varieties. Retrotransposon markers in RBIP format proved to be the most robust and easy to score method, while multilocus IRAP produced informative fingerprint already in a single analysis. Isozyme analysis offered a fast and less expensive alternative. The results showed that molecular identification could be used to assess distinctness and complement morphological assessment, especially in cases where the time frame plays an important role. Currently developed pea marker systems might serve also for germplasm management and genetic diversity studies.

  5. Long-Term Fungal Inhibition by Pisum sativum Flour Hydrolysate during Storage of Wheat Flour Bread

    PubMed Central

    Lavecchia, Anna; Gramaglia, Valerio; Gobbetti, Marco

    2015-01-01

    In order to identify antifungal compounds from natural sources to be used as ingredients in the bakery industry, water/salt-soluble extracts (WSE) from different legume flour hydrolysates obtained by the use of a fungal protease were assayed against Penicillium roqueforti DPPMAF1. The agar diffusion assays allowed the selection of the pea (Pisum sativum) hydrolysate as the most active. As shown by the hyphal radial growth rate, the WSE had inhibitory activity towards several fungi isolated from bakeries. The MIC of the WSE was 9.0 mg/ml. Fungal inhibition was slightly affected by heating and variations in pH. The antifungal activity was attributed to three native proteins (pea defensins 1 and 2 and a nonspecific lipid transfer protein [nsLTP]) and a mixture of peptides released during hydrolysis. The three proteins have been reported previously as components of the defense system of the plant. Five peptides were purified from WSE and were identified as sequences encrypted in leginsulin A, vicilin, provicilin, and the nsLTP. To confirm antifungal activity, the peptides were chemically synthesized and tested. Freeze-dried WSE were used as ingredients in leavened baked goods. In particular, breads made by the addition of 1.6% (wt/wt) of the extract and fermented by baker's yeast or sourdough were characterized for their main chemical, structural, and sensory features, packed in polyethylene bags, stored at room temperature, and compared to controls prepared without pea hydrolysate. Artificially inoculated slices of a bread containing the WSE did not show contamination by fungi until at least 21 days of storage and behaved like the bread prepared with calcium propionate (0.3%, wt/wt). PMID:25862230

  6. Long-Term Fungal Inhibition by Pisum sativum Flour Hydrolysate during Storage of Wheat Flour Bread.

    PubMed

    Rizzello, Carlo Giuseppe; Lavecchia, Anna; Gramaglia, Valerio; Gobbetti, Marco

    2015-06-15

    In order to identify antifungal compounds from natural sources to be used as ingredients in the bakery industry, water/salt-soluble extracts (WSE) from different legume flour hydrolysates obtained by the use of a fungal protease were assayed against Penicillium roqueforti DPPMAF1. The agar diffusion assays allowed the selection of the pea (Pisum sativum) hydrolysate as the most active. As shown by the hyphal radial growth rate, the WSE had inhibitory activity towards several fungi isolated from bakeries. The MIC of the WSE was 9.0 mg/ml. Fungal inhibition was slightly affected by heating and variations in pH. The antifungal activity was attributed to three native proteins (pea defensins 1 and 2 and a nonspecific lipid transfer protein [nsLTP]) and a mixture of peptides released during hydrolysis. The three proteins have been reported previously as components of the defense system of the plant. Five peptides were purified from WSE and were identified as sequences encrypted in leginsulin A, vicilin, provicilin, and the nsLTP. To confirm antifungal activity, the peptides were chemically synthesized and tested. Freeze-dried WSE were used as ingredients in leavened baked goods. In particular, breads made by the addition of 1.6% (wt/wt) of the extract and fermented by baker's yeast or sourdough were characterized for their main chemical, structural, and sensory features, packed in polyethylene bags, stored at room temperature, and compared to controls prepared without pea hydrolysate. Artificially inoculated slices of a bread containing the WSE did not show contamination by fungi until at least 21 days of storage and behaved like the bread prepared with calcium propionate (0.3%, wt/wt). Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  7. Rapidly Induced Wound Ethylene from Excised Segments of Etiolated Pisum sativum L., cv. Alaska

    PubMed Central

    Saltveit, Mikal E.; Dilley, David R.

    1978-01-01

    Wound-induced ethylene synthesis by subapical stem sections of etiolated Pisum sativum L., cv. Alaska seedlings, as described by Saltveit and Dilley (Plant Physiol 1978 61: 447-450), was half-saturated at 3.6% (v/v) O2 and saturated at about 10% O2. Corresponding values for CO2 production during the same period were 1.1% and 10% O2, respectively. Anaerobiosis stopped all ethylene evolution and delayed the characteristic pattern of wound ethylene synthesis. Exposing tissue to 3.5% CO2 in air in a flow-through system reduced wound ethylene synthesis by 30%. Enhancing gas diffusivity by reducing the total pressure to 130 mm Hg almost doubled the rate of wound ethylene synthesis and this effect was negated by exposure to 250 μl liter−1 propylene. Applied ethylene or propylene stopped wound ethylene synthesis during the period of application, but unlike N2, no lag period was observed upon flushing with air. It is concluded that the characteristic pattern of wound-induced ethylene synthesis resulted from negative feedback control by endogenous ethylene. No wound ethylene was produced for 2 hours after excision at 10 or 38 C. Low temperatures prolonged the lag period, but did not prevent induction of the wound response, since tissue held for 2 hours at 10 C produced wound ethylene immediately when warmed to 30 C. In contrast, temperatures above 36 C prevented induction of wound ethylene synthesis, since tissue cooled to 30 C after 1 hour at 40 C required 2 hours before ethylene production returned to normal levels. The activation energy between 15 and 36 C was 12.1 mole kilocalories degree−1. PMID:16660362

  8. Alternative Path Mediated ATP Synthesis in Roots of Pisum sativum upon Nitrogen Supply 1

    PubMed Central

    de Visser, Ries; Brouwer, Koos Spreen; Posthumus, Freek

    1986-01-01

    Changes in the efficiency of root respiration were examined on intact plants of Pisum sativum L. cv Rondo after addition of nitrate or ammonium to the culture solutions. Nitrate was absorbed immediately after addition and elicited a respiratory rise (O2-uptake as well as CO2-production) to 160% at most. This occurred both in roots of plants fixing N2 and in those of non-nodulated plants pregrown for 1 or 2 weeks on a nitrogen-free culture solution. In older plants, used after 2 weeks of N-free growth, the full capacity of the cytochrome path was engaged in root respiration. This was demonstrated by the absence of an effect of the uncoupler carbonylcyanide m-chlorophenylhydrazone in the presence of 25 millimolar salicylhydroxamate, an inhibitor of the alternative path. In these plants more than 90% of the nitrate-induced stimulation of root respiration was salicylhydroxamate-sensitive. In young plants, used after 1 week of N-free growth, the cytochrome path was not saturated. Its activity increased instantaneously at the expense of alternative path activity, which initially dropped to zero and subsequently increased to 160% of the control 7 hours after nitrate supply. The rate of photosynthesis rose to 120% of the control, but not before 1 hour after nitrate supply, suggesting that the stimulation of root respiration was not due to a higher rate of photosynthesis. Experiments with plants grown with a split-root system showed that respiration rate and alternative path activity only increased in the root halves exposed to nitrogen. Ammonium was equally effective as nitrate in stimulating root respiration. These results lead to the conclusion that alternative-path mediated root respiration contributes to synthesis of ATP during at least the first 24 hours following nitrogen supply. Images Fig. 5 PMID:16664616

  9. Carbon Transfer and Partitioning between Vegetative and Reproductive Organs in Pisum sativum L

    PubMed Central

    Jeuffroy, Marie-Hélène; Warembourg, Fernand R.

    1991-01-01

    Assimilate partitioning was studied in the common pea (Pisum sativum L.) by feeding 14CO2 to whole plants and measuring radioactivity in different organs 48 hours after labeling. Two experimental protocols were used. For the first, one reproductive node was darkened with an aluminum foil, to prevent photosynthesis during labeling. The aim was to study assimilate translocation among nodes. The second was carried out to assess any priority among sinks. Whole plants were shaded, during labeling, to reduce carbon assimilation. Various developmental stages between the onset of flowering and the final stage in seed abortion of the last pod were chosen for labeling. When all photosynthetic structures at the first reproductive node were darkened at any stage of development after the formation of the first flower, the first pod was supplied with assimilates from other nodes. In contrast, later developed pods, when photosynthetic structures at their node were darkened, received assimilates from other nodes only when they were beyond their final stage in seed abortion. Reducing illumination to 30% did not change distribution of assimilated carbon between vegetative and reproductive structures, nor among pods. It appears that the relative proportion of 14C allocated to any one pod, compared to other pods, depends on the dry weight of that pod as a proportion of the total reproductive dry weight. When the plant was growing actively, following the start of the reproductive phase until a few days before the end of flowering, the top of the plant (i.e., all the organs above the last opened flower) had a higher sink strength and a higher relative specific activity than pods, suggesting that it was a more competitive sink for assimilates. The pattern of assimilate distribution described here provides an explanation for pod and seed abortion. PMID:16668406

  10. Physiology of Movements in Stems of Seedling Pisum sativum L. cv. Alaska 12

    PubMed Central

    Britz, Steven J.; Galston, Arthur W.

    1982-01-01

    Gravitropism and nutation in the stems of dark-grown, seedling peas (Pisum sativum L. cv. Alaska) were recorded on time-lapse photographs made with photomorphogenetically inactive light. Although gravitropism and nutation have been connected by several different theories in the past, our experiments indicate that the two processes are in fact dissociable. The evidence is as follows: (a) Nutational patterns are asymmetric. There is much greater amplitude of oscillation in the plane parallel (∥) to the plane of the apical hook than in the plane perpendicular (⊥), yet the average gravitropic response is equal in these two planes. (b) Brief red light irradiation given 16 to 24 hours before observation greatly increases the amplitude of nutation in the ∥-plane, but has no influence on the kinetics of gravitropic response. (c) An inhibitor of auxin transport, α-naphthylphthalamic acid, strongly inhibits nutation at 5 micromolar but affects gravitropism only at higher concentrations. (d) Nutation is also strongly inhibited by removal of the apical bud, but gravitropism is unaffected. (e) The period of nutation does not exhibit a constant relationship to the response time of gravitropism. The above evidence is inconsistent with theories that gravitropism is an asymmetrically modified nutation or, alternatively, that nutational oscillations result in a simple fashion from gravitropic overshoots. The evidence is consistent with, although not proof of, autonomous factors such as an endogenous rhythm of growth as the cause of nutation in pea stems. However, gravity and nutation do interact. Nutation in a population of seedlings can be synchronized and brought into phase by a single gravitropic induction. Furthermore, the response time and initial rate of gravitropic curvature depend to some extent on the phase of nutational curvature at which gravitropic induction is begun. PMID:16662458

  11. Bean pod mottle virus: a new powerful tool for functional genomics studies in Pisum sativum.

    PubMed

    Meziadi, Chouaib; Blanchet, Sophie; Richard, Manon M S; Pilet-Nayel, Marie-Laure; Geffroy, Valérie; Pflieger, Stéphanie

    2016-08-01

    Pea (Pisum sativum L.) is an important legume worldwide. The importance of pea in arable rotations and nutritional value for both human and animal consumption have fostered sustained production and different studies to improve agronomic traits of interest. Moreover, complete sequencing of the pea genome is currently underway and will lead to the identification of a large number of genes potentially associated with important agronomic traits. Because stable genetic transformation is laborious for pea, virus-induced gene silencing (VIGS) appears as a powerful alternative technology for determining the function of unknown genes. In this work, we present a rapid and efficient viral inoculation method using DNA infectious plasmids of Bean pod mottle virus (BPMV)-derived VIGS vector. Six pea genotypes with important genes controlling biotic and/or abiotic stresses were found susceptible to BPMV carrying a GFP reporter gene and showed fluorescence in both shoots and roots. In a second step, we investigated 37 additional pea genotypes and found that 30 were susceptible to BPMV and only 7 were resistant. The capacity of BPMV to induce silencing of endogenes was investigated in the most susceptible genotype using two visual reporter genes: PsPDS and PsKORRIGAN1 (PsKOR1) encoding PHYTOENE DESATURASE and a 1,4-β-D-glucanase, respectively. The features of the 'one-step' BPMV-derived VIGS vector include (i) the ease of rub-inoculation, without any need for biolistic or agro-inoculation procedures, (ii) simple cost-effective procedure and (iii) noninterference of viral symptoms with silencing. These features make BPMV the most adapted VIGS vector in pea to make low- to high-throughput VIGS studies. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  12. Antioxidant Enzymes Regulate Reactive Oxygen Species during Pod Elongation in Pisum sativum and Brassica chinensis

    PubMed Central

    Liu, Nan; Lin, Zhifang; Guan, Lanlan; Gaughan, Gerald; Lin, Guizhu

    2014-01-01

    Previous research has focused on the involvement of reactive oxygen species (ROS) in cell wall loosening and cell extension in plant vegetative growth, but few studies have investigated ROS functions specifically in plant reproductive organs. In this study, ROS levels and antioxidant enzyme activities were assessed in Pisum sativum and Brassica chinensis pods at five developmental stages. In juvenile pods, the high levels of O2.− and.OH indicates that they had functions in cell wall loosening and cell elongation. In later developmental stages, high levels of.OH were also related to increases in cell wall thickness in lignified tissues. Throughout pod development, most of the O2.− was detected on plasma membranes of parenchyma cells and outer epidermis cells of the mesocarp, while most of the H2O2 was detected on plasma membranes of most cells throughout the mesocarp. This suggests that these sites are presumably the locations of ROS generation. The antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) apparently contributed to ROS accumulation in pod wall tissues. Furthermore, specifically SOD and POD were found to be associated with pod growth through the regulation of ROS generation and transformation. Throughout pod development, O2.− decreases were associated with increased SOD activity, while changes in H2O2 accumulation were associated with changes in CAT and POD activities. Additionally, high POD activity may contribute to the generation of.OH in the early development of pods. It is concluded that the ROS are produced in different sites of plasma membranes with the regulation of antioxidant enzymes, and that substantial ROS generation and accumulation are evident in cell elongation and cell wall loosening in pod wall cells. PMID:24503564

  13. Physiological effects of nanoparticulate ZnO in green peas (Pisum sativum L.) cultivated in soil.

    PubMed

    Mukherjee, Arnab; Peralta-Videa, Jose R; Bandyopadhyay, Susmita; Rico, Cyren M; Zhao, Lijuan; Gardea-Torresdey, Jorge L

    2014-01-01

    The toxicological effects of zinc oxide nanoparticles (ZnO NPs) in plants are still largely unknown. In the present study, green pea (Pisum sativum L.) plants were treated with 0, 125, 250, and 500 mg kg(-1) of either ZnO NPs or bulk ZnO in organic matter enriched soil. Corresponding toxicological effects were measured on the basis of plant growth, chlorophyll production, Zn bioaccumulation, H2O2 generation, stress enzyme activity, and lipid peroxidation using different cellular, molecular, and biochemical approaches. Compared to control, all ZnO NP concentrations significantly increased (p ≤ 0.05) root elongation but no effects were observed in the stem. Whereas all bulk ZnO treatments significantly increased both root and stem length. After 25 days, chlorophyll in leaves decreased, compared to control, by ~61%, 67%, and 77% in plants treated with 125, 250, and 500 mg kg(-1) ZnO NPs, respectively. Similar results were found in bulk ZnO treated plants. At all ZnO NP concentrations CAT was significantly reduced in leaves (p ≤ 0.05), while APOX was reduced in both roots and leaves. In the case of bulk ZnO, APOX activity was down-regulated in the root and leaf and CAT was unaffected. At 500 mg kg(-1) treatment, the H2O2 in leaves increased by 61% with a twofold lipid peroxidation, which would be a predictive biomarker of nanotoxicity. This study could be pioneering in evaluating the phytotoxicity of ZnO NPs to green peas and can serve as a good indicator for measuring the effects on ZnO NPs in plants grown in organic matter enriched soil.

  14. Characterization of alpha-Amylase from Shoots and Cotyledons of Pea (Pisum sativum L.) Seedlings.

    PubMed

    Beers, E P; Duke, S H

    1990-04-01

    The most abundant alpha-amylase (EC 3.2.1.1) in shoots and cotyledons from pea (Pisum sativum L.) seedlings was purified 6700-and 850-fold, respectively, utilizing affinity (amylose and cycloheptaamylose) and gel filtration chromatography and ultrafiltration. This alpha-amylase contributed at least 79 and 15% of the total amylolytic activity in seedling cotyledons and shoots, respectively. The enzyme was identified as an alpha-amylase by polarimetry, substrate specificity, and end product analyses. The purified alpha-amylases from shoots and cotyledons appear identical. Both are 43.5 kilodalton monomers with pls of 4.5, broad pH activity optima from 5.5 to 6.5, and nearly identical substrate specificities. They produce identical one-dimensional peptide fingerprints following partial proteolysis in the presence of SDS. Calcium is required for activity and thermal stability of this amylase. The enzyme cannot attack maltodextrins with degrees of polymerization below that of maltotetraose, and hydrolysis of intact starch granules was detected only after prolonged incubation. It best utilizes soluble starch as substrate. Glucose and maltose are the major end products of the enzyme with amylose as substrate. This alpha-amylase appears to be secreted, in that it is at least partially localized in the apoplast of shoots. The native enzyme exhibits a high degree of resistance to degradation by proteinase K, trypsin/chymostrypsin, thermolysin, and Staphylococcus aureus V8 protease. It does not appear to be a high-mannose-type glycoprotein. Common cell wall constituents (e.g. beta-glucan) are not substrates of the enzyme. A very low amount of this alpha-amylase appears to be associated with chloroplasts; however, it is unclear whether this activity is contamination or alpha-amylase which is integrally associated with the chloroplast.

  15. Synthesis of Phytochelatins and Homo-Phytochelatins in Pisum sativum L.

    PubMed Central

    Klapheck, S.; Schlunz, S.; Bergmann, L.

    1995-01-01

    In the roots of pea plants (Pisum sativum L.) cultivated with 20 [mu]M CdCl2 for 3 d, synthesis of phytochelatins [PCs or ([gamma]EC)nG, where [gamma]EC is [gamma]glutamylcysteine and G is glycine] and homophytochelatins [h-PCs, ([gamma]EC)n[beta]-alanine] is accompanied by a drastic decrease in glutathione (GSH) content, but an increase in homoglutathione (h-GSH) content. In contrast, the in vitro activity of GSH synthetase increases 5-fold, whereas h-GSH synthetase activity increases regardless of Cd exposure. The consititutive enzyme PC synthase, which catalyzes the transfer of the [gamma]-EC moiety of GSH to an acceptor GSH molecule thus producing ([gamma]EC)2G, is activated by heavy metals, with Cd and Cu being strong activators and Zn being a very poor activator. Using h-GSH or hm-GSH for substrate, the synthesis rate of([gamma]EC)2[beta]-alanine and [gamma]EC)2-serine is only 2.4 and 0.3%, respectively, of the sythesis rate of ([gamma]EC)2G with GSH as substrate. However, in the presence of a constant GSH level, increasing the concentration of h-GSH or hm-GSH results in increased synthesis of ([gamma]EC)2[beta]-alanine or ([gamma]EC)2-serine, respecively; simultaneously, the synthesis of ([gamma]EC)2G is inhibited. [gamma]EC is not a substrate of PC synthase. These results are best explained by assuming that PC synthase has a [gamma]EC donor binding site, which is very specific for GSH, and a [gamma]EC acceptor binding site, which is less specific and accepts several tripeptides, namely GSH, h-GSH, and hm-GSH. PMID:12228379

  16. Sitona lineatus (Coleoptera: Curculionidae) Larval Feeding on Pisum sativum L. Affects Soil and Plant Nitrogen

    PubMed Central

    Cárcamo, Héctor A.; Herle, Carolyn E.; Lupwayi, Newton Z.

    2015-01-01

    Adults of Sitona lineatus (pea leaf weevil, PLW) feed on foliage of several Fabaceae species but larvae prefer to feed on nodules of Pisum sativum L. and Vicia faba L. Indirectly, through their feeding on rhizobia, weevils can reduce soil and plant available nitrogen (N). However, initial soil N can reduce nodulation and damage by the weevil and reduce control requirements. Understanding these interactions is necessary to make integrated pest management recommendations for PLW. We conducted a greenhouse study to quantify nodulation, soil and plant N content, and nodule damage by weevil larvae in relation to soil N amendment with urea, thiamethoxam insecticide seed coating and crop stage. PLWs reduced the number of older tumescent (multilobed) nodules and thiamethoxam addition increased them regardless of other factors. Nitrogen amendment significantly increased soil available N (>99% nitrate) as expected and PLW presence was associated with significantly lower levels of soil N. PLW decreased plant N content at early flower and thiamethoxam increased it, particularly at late flower. The study illustrated the complexity of interactions that determine insect herbivory effects on plant and soil nutrition for invertebrates that feed on N-fixing root nodules. We conclude that effects of PLW on nodulation and subsequent effects on plant nitrogen are more pronounced during the early growth stages of the plant. This suggests the importance of timing of PLW infestation and may explain the lack of yield depression in relation to this pest observed in many field studies. Also, pea crops in soils with high levels of soil N are unlikely to be affected by this herbivore and should not require insecticide inputs. PMID:26106086

  17. Molecular changes in Pisum sativum L. roots during arbuscular mycorrhiza buffering of cadmium stress.

    PubMed

    Rivera-Becerril, Facundo; van Tuinen, Diederik; Martin-Laurent, Fabrice; Metwally, Ashraf; Dietz, Karl-Josef; Gianinazzi, Silvio; Gianinazzi-Pearson, Vivienne

    2005-12-01

    Molecular responses to cadmium (Cd) stress were studied in mycorrhizal and non-mycorrhizal Pisum sativum L. cv. Frisson inoculated with Glomus intraradices. Biomass decreases caused by the heavy metal were significantly less in mycorrhizal than in non-mycorrhizal plants. Real-time reverse transcriptase-polymerase chain reaction showed that genes implicated in pathways of Cd detoxification varied in response to mycorrhiza development or Cd application. Expression of a metallothionein-encoding gene increased strongly in roots of Cd-treated non-mycorrhizal plants. Genes encoding gamma-glutamylcysteine synthetase and glutathione (GSH) synthetase, responsible for the synthesis of the phytochelatin (PC) precursor GSH, were activated by Cd in mycorrhizal and non-mycorrhizal plants. Cd stress decreased accumulation of GSH/homoglutathione (hGSH) and increased thiol groups in pea roots, whether mycorrhizal or not, suggesting synthesis of PCs and/or homophytochelatins. An hGSH synthetase gene, involved in hGSH synthesis, did not respond to Cd alone but was activated by mycorrhizal development in the presence of Cd. Transcript levels of a glutathione reductase gene were only increased in non-mycorrhizal roots treated with Cd. Studies of three stress-related genes showed that a heat-shock protein gene was activated in mycorrhizal roots or by Cd and chitinase gene transcripts increased under Cd stress to a greater extent in mycorrhizal roots, whilst a chalcone isomerase gene was only up-regulated by Cd. Results indicate that although heavy metal chelation pathways contribute to Cd stress responses in pea, they may not make a major contribution to Cd tolerance strategies operating in the arbuscular mycorrhizal symbiosis.

  18. Kinetic properties of a micronutrient transporter from Pisum sativum indicate a primary function in Fe uptake from the soil.

    PubMed

    Cohen, Clara K; Garvin, David F; Kochian, Leon V

    2004-03-01

    Fe uptake in dicotyledonous plants is mediated by a root plasma membrane-bound ferric reductase that reduces extracellular Fe(III)-chelates, releasing Fe(2+) ions, which are then absorbed via a metal ion transporter. We previously showed that Fe deficiency induces an increased capacity to absorb Fe and other micronutrient and heavy metals such as Zn(2+) and Cd(2+) into pea ( Pisum sativum L.) roots [Cohen et al. (1998) Plant Physiol 116:1063-1072). To investigate the molecular basis for this phenomenon, an Fe-regulated transporter that is a homologue of the Arabidopsis IRT1 micronutrient transporter was isolated from pea seedlings. This cDNA clone, designated RIT1 for root iron transporter, encodes a 348 amino acid polypeptide with eight putative membrane-spanning domains that is induced under Fe deficiency and can functionally complement yeast mutants defective in high- and low-affinity Fe transport. Chelate buffer techniques were used to control Fe(2+) in the uptake solution at nanomolar activities representative of those found in the rhizosphere, and radiotracer methodologies were employed to show that RIT1 is a very high-affinity (59)Fe(2+) uptake system ( K(m) =54-93 nM). Additionally, radiotracer ((65)Zn, (109)Cd) flux techniques were used to show that RIT can also mediate a lower affinity Zn and Cd influx ( K(m) of 4 and 100 microM, for Zn(2+) and Cd(2+), respectively). These findings suggest that, in typical agricultural soils, RIT1 functions primarily as a high-affinity Fe(2+) transporter that mediates root Fe acquisition. This is consistent with recent findings with Arabidopsis IRT1 knockout mutants that strongly suggest that this transporter plays a key role in root Fe uptake and nutrition. However, the ability of RIT1 to facilitate Zn and Cd uptake when these metals are present at elevated concentrations suggests that RIT1 may be one pathway for the entry of toxic metals into the food chain. Furthermore, the finding that plant Fe deficiency status may

  19. Physical basis for altered stem elongation rates in internode length mutants of Pisum

    NASA Technical Reports Server (NTRS)

    Behringer, F. J.; Cosgrove, D. J.; Reid, J. B.; Davies, P. J.

    1990-01-01

    Biophysical parameters related to gibberellin (GA)-dependent stem elongation were examined in dark-grown stem-length genotypes of Pisum sativum L. The rate of internode expansion in these genotypes is altered due to recessive mutations which affect either the endogenous levels of, or response to, GA. The GA deficient dwarf L181 (ls), two GA insensitive semierectoides dwarfs NGB5865 and NGB5862 (lka and lkb, respectively) and the slender' line L197 (la crys), which is tall regardless of GA content, were compared to the wild-type tall cultivar, Torsdag. Osmotic pressure, estimated by vapor pressure osmometry, and turgor pressure, measured directly with a pressure probe, did not correlate with the differences in growth rate among the genotypes. Mechanical wall properties of frozen-thawed tissue were measured using a constant force assay. GA deficiency resulted in increased wall stiffness judged both on the basis of plastic compliance and plastic extensibility normalized for equal stem circumference. Plastic compliance was not reduced in the GA insensitive dwarfs, though lka reduced circumference-normalized plasticity. In contrast, in vivo wall relaxation, determined by the pressure-block technique, differed among genotypes in a manner which did correlate with extension rates. The wall yield threshold was 1 bar or less in the tall lines, but ranged from 3 to 6 bars in the dwarf genotypes. The results with the ls mutant indicate that GA enhances stem elongation by both decreasing the wall yield threshold and increasing the wall yield coefficient. In the GA-insensitive mutants, lka and lkb, the wall yield threshold is substantially elevated. Plants possessing lka may also possess a reduced wall yield coefficient.

  20. Physical basis for altered stem elongation rates in internode length mutants of Pisum

    SciTech Connect

    Behringer, F.J.; Davies, P.J. ); Cosgrove, D.J. ); Reid, J.B. )

    1990-09-01

    Biophysical parameters related to gibberellin (GA)-dependent stem elongation were examined in dark-grown stem-length genotypes of Pisum sativum L. The rate of internode expansion in these genotypes is altered due to recessive mutations which affect either the endogenous levels of, or response to, GA. The GA deficient dwarf L181 (ls), two GA insensitive semierectoides dwarfs NGB5865 and NGB5862 (lka and lkb, respectively) and the slender line L197 (la cry{sup s}), which is tall regardless of GA content, were compared to the wild-type tall cultivar, Torsdag. Osmotic pressure, estimated by vapor pressure osmometry, and turgor pressure, measured directly with a pressure probe, did not correlate with the differences in growth rate among the genotypes. Mechanical wall properties of frozen-thawed tissue were measured using a constant force assay. GA deficiency resulted in increased wall stiffness judged both on the basis of plastic compliance and plastic extensibility normalized for equal stem circumference. Plastic compliance was not reduced in the GA insensitive dwarfs, though lka reduced circumference-normalized plasticity. In contrast, in vivo wall relaxation, determined by the pressure-block technique, differed among genotypes in a manner which did correlate with extension rates. The wall yield threshold was 1 bar or less in the tall lines, but ranged from 3 to 6 bars in the dwarf genotypes. The results with the ls mutant indicate that GA enhances stem elongation by both decreasing the wall yield threshold and increasing the wall yield coefficient. In the GA-insensitive mutants, lka and lkb, the wall yield threshold is substantially elevated. Plants possessing lka may also possess a reduced wall yield coefficient.

  1. Physical basis for altered stem elongation rates in internode length mutants of Pisum.

    PubMed

    Behringer, F J; Cosgrove, D J; Reid, J B; Davies, P J

    1990-01-01

    Biophysical parameters related to gibberellin (GA)-dependent stem elongation were examined in dark-grown stem-length genotypes of Pisum sativum L. The rate of internode expansion in these genotypes is altered due to recessive mutations which affect either the endogenous levels of, or response to, GA. The GA deficient dwarf L181 (ls), two GA insensitive semierectoides dwarfs NGB5865 and NGB5862 (lka and lkb, respectively) and the slender' line L197 (la crys), which is tall regardless of GA content, were compared to the wild-type tall cultivar, Torsdag. Osmotic pressure, estimated by vapor pressure osmometry, and turgor pressure, measured directly with a pressure probe, did not correlate with the differences in growth rate among the genotypes. Mechanical wall properties of frozen-thawed tissue were measured using a constant force assay. GA deficiency resulted in increased wall stiffness judged both on the basis of plastic compliance and plastic extensibility normalized for equal stem circumference. Plastic compliance was not reduced in the GA insensitive dwarfs, though lka reduced circumference-normalized plasticity. In contrast, in vivo wall relaxation, determined by the pressure-block technique, differed among genotypes in a manner which did correlate with extension rates. The wall yield threshold was 1 bar or less in the tall lines, but ranged from 3 to 6 bars in the dwarf genotypes. The results with the ls mutant indicate that GA enhances stem elongation by both decreasing the wall yield threshold and increasing the wall yield coefficient. In the GA-insensitive mutants, lka and lkb, the wall yield threshold is substantially elevated. Plants possessing lka may also possess a reduced wall yield coefficient.

  2. Physical basis for altered stem elongation rates in internode length mutants of Pisum

    NASA Technical Reports Server (NTRS)

    Behringer, F. J.; Cosgrove, D. J.; Reid, J. B.; Davies, P. J.

    1990-01-01

    Biophysical parameters related to gibberellin (GA)-dependent stem elongation were examined in dark-grown stem-length genotypes of Pisum sativum L. The rate of internode expansion in these genotypes is altered due to recessive mutations which affect either the endogenous levels of, or response to, GA. The GA deficient dwarf L181 (ls), two GA insensitive semierectoides dwarfs NGB5865 and NGB5862 (lka and lkb, respectively) and the slender' line L197 (la crys), which is tall regardless of GA content, were compared to the wild-type tall cultivar, Torsdag. Osmotic pressure, estimated by vapor pressure osmometry, and turgor pressure, measured directly with a pressure probe, did not correlate with the differences in growth rate among the genotypes. Mechanical wall properties of frozen-thawed tissue were measured using a constant force assay. GA deficiency resulted in increased wall stiffness judged both on the basis of plastic compliance and plastic extensibility normalized for equal stem circumference. Plastic compliance was not reduced in the GA insensitive dwarfs, though lka reduced circumference-normalized plasticity. In contrast, in vivo wall relaxation, determined by the pressure-block technique, differed among genotypes in a manner which did correlate with extension rates. The wall yield threshold was 1 bar or less in the tall lines, but ranged from 3 to 6 bars in the dwarf genotypes. The results with the ls mutant indicate that GA enhances stem elongation by both decreasing the wall yield threshold and increasing the wall yield coefficient. In the GA-insensitive mutants, lka and lkb, the wall yield threshold is substantially elevated. Plants possessing lka may also possess a reduced wall yield coefficient.

  3. High-throughput development of SSR markers from pea (Pisum sativum L.) based on next generation sequencing of a purified Chinese commercial variety

    USDA-ARS?s Scientific Manuscript database

    Pea (Pisum sativum L.) is an important food legume globally, and is the plant species that J.G. Mendel used to lay the foundation of modern genetics. However, genomics resources of pea are limited comparing to other crop species. Application of marker assisted selection (MAS) in pea breeding has lag...

  4. Genetic diversity, population structure and genome-wide marker-trait association analysis of the USDA pea (Pisum sativum L.) core collection

    USDA-ARS?s Scientific Manuscript database

    Genetic diversity, population structure and genome-wide marker-trait association analysis was conducted for the USDA pea (Pisum sativum L.) core collection. The core collection contained 285 accessions with diverse phenotypes and geographic origins. The 137 DNA markers included 102 polymorphic fra...

  5. Large-scale evaluation of pea (Pisum sativum L.) germplasm for cold tolerance in the open field during winter in Qingdao.

    USDA-ARS?s Scientific Manuscript database

    As a cool season crop, pea (Pisum sativum L.) can tolerate frost at the vegetative stage but has yield loss when freezing stress occurs at reproductive stage. Cold tolerance improvement of pea varieties is important for the stable yield and the expansion of winter pea planting area. Under the natura...

  6. Evaluation of expression stability of candidate references genes among green and yellow pea cultivars (Pisum sativum L.) subjected to abiotic and biotic stress

    USDA-ARS?s Scientific Manuscript database

    Dry pea (Pisum sativum) is grown as human and animal feed throughout the world. Large yield losses in pea due to biotic and abiotic stresses compel an improved understanding of mechanisms of stress tolerance and genetic determinants conditioning these tolerances. The availability of stably expressed...

  7. Complete Release of Axillary Buds from Apical Dominance in Intact, Light-Grown Seedlings of Pisum sativum L. following a Single Application of Cytokinin 1

    PubMed Central

    Pillay, Indiren; Railton, Ian D.

    1983-01-01

    Single applications of either 6-benzyladenine or zeatin to inhibited axillary buds of intact, light-grown seedlings of Pisum sativum L. cv Black-eyed Susan, resulted in the formation of rapidly elongating, leafy shoots. Similar treatment with kinetin or isopentenyladenine caused only limited but outgrowth which stopped 6 days after application. Images Fig. 2 PMID:16662939

  8. Development and Partial Characterization of Nearly Isogenic Pea Lines (Pisum sativum L.) that Alter Uptake Hydrogenase Activity in Symbiotic Rhizobium.

    PubMed

    Phillips, D A; Kapulnik, Y; Bedmar, E J; Joseph, C M

    1990-04-01

    Some Rhizobium bacteria have H(2)-uptake (Hup) systems that oxidize H(2) evolved from nitrogenase in leguminous root nodules. Pea (Pisum sativum L.) cultivars ;JI1205' and ;Alaska' produce high Hup (Hup(++)) and moderate Hup (Hup(+)) phenotypes, respectively, in Rhizobium leguminosarum 128C53. The physiological significance and biochemical basis of this host plant genetic effect are unknown. The purpose of this investigation was to advance basic Hup studies by developing nearly isogenic lines of peas that alter Hup phenotypes in R. leguminosarum strains containing hup genes. Eight pairs of nearly isogenic pea lines that produce Hup(++) and Hup(+) phenotypes in R. leguminosarum 128C53 were identified in 173 F(2)-derived F(6) families produced from crosses between JI1205 and Alaska. Tests with the pea isolines and three strains of hup-containing R. leguminosarum showed that the isolines altered Hup activity significantly (P mutants of R. leguminosarum 128C53 showed that, in symbioses with Hup(+) rhizobia, two out of three Hup(++) pea lines decreased N(2) fixation relative to Hup(+) peas. In one of those cases, however, the Hup(++) plant line also decreased fixation by Hup(-) rhizobia. When results were averaged across all rhizobia tested, Hup(+) pea isolines had 8.2% higher dry weight (P

  9. Stomatal Response and Leaf Injury of Pisum sativum L. with SO2 and O3 Exposures 12

    PubMed Central

    Olszyk, David M.; Tibbitts, Theodore W.

    1981-01-01

    Stomatal response during exposure to SO2 and O3 and subsequent leaf injury were examined in plants of Pisum sativum L. `Alsweet' grown in a peat-vermiculite medium in controlled environment chambers. Plants developing under moisture stress, induced by drying the medium to 50% of field capacity, exhibited greater stomatal closure during exposures and less than one-fourth the necrosis compared to plants developing in a medium maintained at field capacity. Plants under moisture stress had only a slightly more negative plant water potential (≃−4.0 bars) than at field capacity (≃−3.4 bars). Plants exposed to pollutants for 2 hours near the beginning or end of a 16-hour light period had greater stomatal closure during exposures and less leaf necrosis than plants exposed during the middle of the light period. Images PMID:16661711

  10. Micromonospora ureilytica sp. nov., Micromonospora noduli sp. nov. and Micromonospora vinacea sp. nov., isolated from Pisum sativum nodules.

    PubMed

    Carro, Lorena; Riesco, Raúl; Spröer, Cathrin; Trujillo, Martha E

    2016-09-01

    A diversity study on the presence of strains representing the genus Micromonospora in Pisum sativum nodules collected from Cañizal (Spain) has provided evidence of the high number of isolates that might represent novel species. In the present work, we have characterized three of these isolates: GUI23T, GUI43T and GUI63T. Phenotypic and genotypic analyses confirmed that all strains represent novel species of the genus Micromonospora with the following proposed names: Micromonospora ureilytica sp. nov., type strain GUI23T (=CECT 9022T=DSM 101692T), Micromonospora noduli sp. nov., type strain GUI43T (=CECT 9020T=DSM 101694T), and Micromonospora vinacea sp. nov., type strain GUI63T (=CECT 9019T=DSM 101695T).

  11. Transgenic peas (Pisum sativum) expressing polygalacturonase inhibiting protein from raspberry (Rubus idaeus) and stilbene synthase from grape (Vitis vinifera).

    PubMed

    Richter, A; Jacobsen, H-J; de Kathen, A; de Lorenzo, G; Briviba, K; Hain, R; Ramsay, G; Kiesecker, H

    2006-11-01

    The pea (Pisum sativum L.) varieties Baroness (United Kingdome) and Baccara (France) were transformed via Agrobacterium tumefaciens-mediated gene transfer with pGPTV binary vectors containing the bar gene in combination with two different antifungal genes coding for polygalacturonase-inhibiting protein (PGIP) from raspberry (Rubus idaeus L.) driven by a double 35S promoter, or the stilbene synthase (Vst1) from grape (Vitis vinifera L.) driven by its own elicitor-inducible promoter. Transgenic lines were established and transgenes combined via conventional crossing. Resveratrol, produced by Vst1 transgenic plants, was detected using HPLC and the PGIP expression was determined in functional inhibition assays against fungal polygalacturonases. Stable inheritance of the antifungal genes in the transgenic plants was demonstrated.

  12. Differential changes in size distribution of xyloglucan in the cell walls of gravitropically responding Pisum sativum epicotyls

    NASA Technical Reports Server (NTRS)

    Talbott, L. D.; Pickard, B. G.

    1994-01-01

    Growth-related change in the size distribution of hemicellulosic wall polymers during the gravitropic curvature response of intact pea (Pisum sativum L. cv Alaska) epicotyls was examined by gel-filtration chromatography. The gravitropic response was characterized by the appearance of curvature 20 to 30 min after horizontal placement, with 35 degrees of curvature attained by 80 min. Correlated with the onset of curvature, on the upper side of the epicotyl, there was a conspicuous transient increase in the abundance of relatively large hemicellulosic xyloglucan polymers, similar to increases previously found under conditions where diminished wall extensibility was expected. On the lower side there was a moderate, slower, and longer-term increase in abundance of small xyloglucan, similar to changes previously found in connection with auxin-stimulated growth responses. Both shifts occurred primarily in the epidermis. They appear to represent two coordinated physiological mechanisms contributing to differential growth.

  13. Differential changes in size distribution of xyloglucan in the cell walls of gravitropically responding Pisum sativum epicotyls.

    PubMed

    Talbott, L D; Pickard, B G

    1994-10-01

    Growth-related change in the size distribution of hemicellulosic wall polymers during the gravitropic curvature response of intact pea (Pisum sativum L. cv Alaska) epicotyls was examined by gel-filtration chromatography. The gravitropic response was characterized by the appearance of curvature 20 to 30 min after horizontal placement, with 35 degrees of curvature attained by 80 min. Correlated with the onset of curvature, on the upper side of the epicotyl, there was a conspicuous transient increase in the abundance of relatively large hemicellulosic xyloglucan polymers, similar to increases previously found under conditions where diminished wall extensibility was expected. On the lower side there was a moderate, slower, and longer-term increase in abundance of small xyloglucan, similar to changes previously found in connection with auxin-stimulated growth responses. Both shifts occurred primarily in the epidermis. They appear to represent two coordinated physiological mechanisms contributing to differential growth.

  14. Nucleotide sequence of a complementary DNA encoding pea cytosolic copper/zinc superoxide dismutase. [Pisum sativum L

    SciTech Connect

    White, D.A.; Zilinskas, B.A. )

    1991-08-01

    The authors now report the nucleotide sequence of the cytosolic Cu/Zn SOD cloned from a {lambda}gt11 cDNA library constructed from mRNA extracted from leaves of 7- to 10-d pea seedlings (Pisum sativum L.). The clone was isolated using a 22-base synthetic oligonucleotide complementary to the amino acid sequence CGIIGLQG. This sequence, found at the protein's carboxy terminus, is highly conserved among plant cytosolic Cu/Zn SODs but not chloroplastic Cu/Zn SODs. The 738-base pair sequence contains an open reading frame specifying 152 codons and a predicted M{sub r} of 18,024 D. The deduced amino acid sequence is highly homologous (79-82% identity) with the sequences of other known plant cytosolic Cu/Zn SODs but less highly conserved (63-65%) when compared with several chloroplastic Cu/Zn SODs including pea (10).

  15. Hypolipidemic Effect of the Autoclaved Extract Prepared from Pea (Pisum sativum L.) Pods In Vivo and In Vitro.

    PubMed

    Inagaki, Kae; Nishimura, Yuuki; Iwata, Emiko; Manabe, Sachinobu; Goto, Masahiro; Ogura, Yoshio; Hotta, Hisako

    2016-01-01

    By autoclaving, we obtained a polyphenol and dietary fiber from pea (Pisum sativum L.) pods in parallel without acid or alkali treatment or organic solvent extraction. Rats fed a high-sucrose (HS) diet containing 3% autoclaved extract (AE) for 4 wk exhibited significantly lower serum triglyceride and total cholesterol levels than rats fed a HS diet. AE and soluble dietary fiber (SDF) from AE exhibited pancreatic lipase inhibitory activity at 13.3 mg/mL in vitro. AE and insoluble dietary fiber (IDF) from AE adsorbed cholesterol. In total, 30% and 10% of a cholesterol micelle were significantly adsorbed by 2,000 mg of AE and 100 mg of IDF from AE in 7 mL, respectively. The amount of bifidobacteria in the cecum of the AE group was significantly increased compared with that in the HS group. These results suggest that AE has hypolipidemic, bifidogenic potential.

  16. Dissipation of pendimethalin in the soil of field pea (Pisum sativum L.) and detection of terminal residues in plants.

    PubMed

    Sondhia, Shobha

    2013-01-01

    Dissipation of pendimethalin in the soil of field peas (Pisum sativum L.) at 0 to 110 days, and terminal residues in green and mature pea were studied under field conditions. Pendimethalin was applied as pre-emergence herbicide at 750, to 185 g a.i. ha(-1) in winter, in field peas. Dissipation of pendimethalin in the soil at 0 to 110 days followed first-order kinetics showing a half-life of 19.83 days averaged over all doses. Low pendimethalin residues were found in mature pea grain (0.004, 0.003, <0.001 μg g(-1)), and straw (0.007, 0.002, <0.001 μg g(-1)) at 750, 350 and 185 g a.i. ha(-1) treatments, respectively. The study indicated that residues of pendimethalin in green and mature pea were within the prescribed MRL limits.

  17. Differential changes in size distribution of xyloglucan in the cell walls of gravitropically responding Pisum sativum epicotyls

    NASA Technical Reports Server (NTRS)

    Talbott, L. D.; Pickard, B. G.

    1994-01-01

    Growth-related change in the size distribution of hemicellulosic wall polymers during the gravitropic curvature response of intact pea (Pisum sativum L. cv Alaska) epicotyls was examined by gel-filtration chromatography. The gravitropic response was characterized by the appearance of curvature 20 to 30 min after horizontal placement, with 35 degrees of curvature attained by 80 min. Correlated with the onset of curvature, on the upper side of the epicotyl, there was a conspicuous transient increase in the abundance of relatively large hemicellulosic xyloglucan polymers, similar to increases previously found under conditions where diminished wall extensibility was expected. On the lower side there was a moderate, slower, and longer-term increase in abundance of small xyloglucan, similar to changes previously found in connection with auxin-stimulated growth responses. Both shifts occurred primarily in the epidermis. They appear to represent two coordinated physiological mechanisms contributing to differential growth.

  18. The effect of propionic acid and valeric acid on the cell cycle in root meristems of Pisum sativum

    SciTech Connect

    Tramontano, W.A.; Yang, Shauyu; Delillo, A.R. )

    1990-01-01

    Propionic acid and valeric acid at 1mM reduced the mitotic index of root meristem cells of Pisum sativum to < 1% after 12 hr in aerated White's medium. This effect varied with different acid concentrations. After a 12 hr exposure to either acid, seedlings transferred to fresh medium without either acid, resumed their normal mitotic index after 12 hr, with a burst of mitosis 8 hr post-transfer. Exposure of root meristem cells to either acid also inhibited ({sup 3}H)-TdR incorporation. Neither acid significantly altered the distribution of meristematic cells in G1 and G2 after 12 hr. The incorporation of ({sup 3}H) - uridine was also unaltered by the addition of either acid. This information suggests that propionic acid and valeric acid, limit progression through the cell cycle by inhibiting DNA synthesis and arresting cells in G1 and G2. These results were consistent with previous data which utilized butyric acid.

  19. The conformational stability and biophysical properties of the eukaryotic thioredoxins of Pisum sativum are not family-conserved.

    PubMed

    Aguado-Llera, David; Martínez-Gómez, Ana Isabel; Prieto, Jesús; Marenchino, Marco; Traverso, José Angel; Gómez, Javier; Chueca, Ana; Neira, José L

    2011-02-22

    Thioredoxins (TRXs) are ubiquitous proteins involved in redox processes. About forty genes encode TRX or TRX-related proteins in plants, grouped in different families according to their subcellular localization. For instance, the h-type TRXs are located in cytoplasm or mitochondria, whereas f-type TRXs have a plastidial origin, although both types of proteins have an eukaryotic origin as opposed to other TRXs. Herein, we study the conformational and the biophysical features of TRXh1, TRXh2 and TRXf from Pisum sativum. The modelled structures of the three proteins show the well-known TRX fold. While sharing similar pH-denaturations features, the chemical and thermal stabilities are different, being PsTRXh1 (Pisum sativum thioredoxin h1) the most stable isoform; moreover, the three proteins follow a three-state denaturation model, during the chemical-denaturations. These differences in the thermal- and chemical-denaturations result from changes, in a broad sense, of the several ASAs (accessible surface areas) of the proteins. Thus, although a strong relationship can be found between the primary amino acid sequence and the structure among TRXs, that between the residue sequence and the conformational stability and biophysical properties is not. We discuss how these differences in the biophysical properties of TRXs determine their unique functions in pea, and we show how residues involved in the biophysical features described (pH-titrations, dimerizations and chemical-denaturations) belong to regions involved in interaction with other proteins. Our results suggest that the sequence demands of protein-protein function are relatively rigid, with different protein-binding pockets (some in common) for each of the three proteins, but the demands of structure and conformational stability per se (as long as there is a maintained core), are less so.

  20. Cell Wall Pectin and its Methyl-esterification in Transition Zone Determine Al Resistance in Cultivars of Pea (Pisum sativum)

    PubMed Central

    Li, Xuewen; Li, Yalin; Qu, Mei; Xiao, Hongdong; Feng, Yingming; Liu, Jiayou; Wu, Lishu; Yu, Min

    2016-01-01

    The initial response of plants to aluminum (Al) is the inhibition of root elongation, while the transition zone is the most Al sensitive zone in the root apex, which may sense the presence of Al and regulate the responses of root to Al toxicity. In the present study, the effect of Al treatment (30 μM, 24 h) on root growth, Al accumulation, and properties of cell wall of two pea (Pisum sativum L.) cultivars, cv Onward (Al-resistant) and cv Sima (Al-sensitive), were studied to disclose whether the response of root transition zone to Al toxicity determines Al resistance in pea cultivars. The lower relative root elongation (RRE) and higher Al content were founded in cv Sima compared with cv Onward, which were related to Al-induced the increase of pectin in root segments of both cultivars. The increase of pectin is more prominent in Al-sensitive cultivar than in Al-resistant cultivar. Aluminum toxicity also induced the increase of pectin methylesterases (PME), which is 2.2 times in root transition zone in Al-sensitive cv Sima to that of Al resistant cv Onward, thus led to higher demethylesterified pectin content in root transition zone of Al-sensitive cv Sima. The higher demethylesterified pectin content in root transition zone resulted in more Al accumulation in the cell wall and cytosol in Al-sensitive cv Sima. Our results provide evidence that the increase of pectin content and PME activity under Al toxicity cooperates to determine Al sensitivity in root transition zone that confers Al resistance in cultivars of pea (Pisum sativum). PMID:26870060

  1. Chromium (VI) induced phytotoxicity and oxidative stress in pea (Pisum sativum L.): biochemical changes and translocation of essential nutrients.

    PubMed

    Tiwari, K K; Dwivedi, S; Singh, N K; Rai, U N; Tripathi, R D

    2009-05-01

    Due to widespread industrial use, chromium (Cr) is considered a hazardous environmental pollutant. It is known to inhibit plant growth and development. The present study provides the evidence of the phytotoxicity of this metal on the pea (Pisum sativum L. cv Azad) plants. The plants of pea (Pisum sativum L.) were grown in refined sand under different concentrations i.e. 0.05, 0.1, 0.2, 0.3 and 0.4 mM of Cr (VI) in order to study the effect on growth and yield, photosynthetic pigments, relative water content, non-reducing sugar and protein with activity of certain enzymes like catalase, peroxidase, starch phosphorylase and ribonuclease. The analysis of the results showed that photosynthetic pigments (68.68%), relative water contents (62.77%), non-reducing sugar (66.66%) and protein (81.57%) were decrease along with reduction in plant height (52.69%) and leaf area (50.81%) of the pea plants. However, in response to various concentration of Cr exposed plants showed significant induction of reducing and total sugars with enzymes like catalase, starch phosphorylase and ribonuclease. The translocation of Cr in various part of pea plant have been found in order of root> stem> leaves>seeds which ranged between 34.8 to 217.3 mg g(-1) d.wt. (dry weight) in roots, 6.5 to 173.13 mg g(-1) d.wt. in shoot, 4.2 to 74.43 mg g(-1) d.wt. in leaves and 0.94 to 8.64 mg g(-1) d.wt. in seeds, that is also reflected by the transfer factor of Cr from refined sand to tested species.

  2. Cell Wall Pectin and its Methyl-esterification in Transition Zone Determine Al Resistance in Cultivars of Pea (Pisum sativum).

    PubMed

    Li, Xuewen; Li, Yalin; Qu, Mei; Xiao, Hongdong; Feng, Yingming; Liu, Jiayou; Wu, Lishu; Yu, Min

    2016-01-01

    The initial response of plants to aluminum (Al) is the inhibition of root elongation, while the transition zone is the most Al sensitive zone in the root apex, which may sense the presence of Al and regulate the responses of root to Al toxicity. In the present study, the effect of Al treatment (30 μM, 24 h) on root growth, Al accumulation, and properties of cell wall of two pea (Pisum sativum L.) cultivars, cv Onward (Al-resistant) and cv Sima (Al-sensitive), were studied to disclose whether the response of root transition zone to Al toxicity determines Al resistance in pea cultivars. The lower relative root elongation (RRE) and higher Al content were founded in cv Sima compared with cv Onward, which were related to Al-induced the increase of pectin in root segments of both cultivars. The increase of pectin is more prominent in Al-sensitive cultivar than in Al-resistant cultivar. Aluminum toxicity also induced the increase of pectin methylesterases (PME), which is 2.2 times in root transition zone in Al-sensitive cv Sima to that of Al resistant cv Onward, thus led to higher demethylesterified pectin content in root transition zone of Al-sensitive cv Sima. The higher demethylesterified pectin content in root transition zone resulted in more Al accumulation in the cell wall and cytosol in Al-sensitive cv Sima. Our results provide evidence that the increase of pectin content and PME activity under Al toxicity cooperates to determine Al sensitivity in root transition zone that confers Al resistance in cultivars of pea (Pisum sativum).

  3. The Conformational Stability and Biophysical Properties of the Eukaryotic Thioredoxins of Pisum Sativum Are Not Family-Conserved

    PubMed Central

    Aguado-Llera, David; Martínez-Gómez, Ana Isabel; Prieto, Jesús; Marenchino, Marco; Traverso, José Angel; Gómez, Javier; Chueca, Ana; Neira, José L.

    2011-01-01

    Thioredoxins (TRXs) are ubiquitous proteins involved in redox processes. About forty genes encode TRX or TRX-related proteins in plants, grouped in different families according to their subcellular localization. For instance, the h-type TRXs are located in cytoplasm or mitochondria, whereas f-type TRXs have a plastidial origin, although both types of proteins have an eukaryotic origin as opposed to other TRXs. Herein, we study the conformational and the biophysical features of TRXh1, TRXh2 and TRXf from Pisum sativum. The modelled structures of the three proteins show the well-known TRX fold. While sharing similar pH-denaturations features, the chemical and thermal stabilities are different, being PsTRXh1 (Pisum sativum thioredoxin h1) the most stable isoform; moreover, the three proteins follow a three-state denaturation model, during the chemical-denaturations. These differences in the thermal- and chemical-denaturations result from changes, in a broad sense, of the several ASAs (accessible surface areas) of the proteins. Thus, although a strong relationship can be found between the primary amino acid sequence and the structure among TRXs, that between the residue sequence and the conformational stability and biophysical properties is not. We discuss how these differences in the biophysical properties of TRXs determine their unique functions in pea, and we show how residues involved in the biophysical features described (pH-titrations, dimerizations and chemical-denaturations) belong to regions involved in interaction with other proteins. Our results suggest that the sequence demands of protein-protein function are relatively rigid, with different protein-binding pockets (some in common) for each of the three proteins, but the demands of structure and conformational stability per se (as long as there is a maintained core), are less so. PMID:21364950

  4. Amino acid fingerprint in the rhizosphere of Pisum sativum in response to water stress

    NASA Astrophysics Data System (ADS)

    Bobille, Hélène; Fustec, Joëlle; Robins, Richard J.; Cukier, Caroline; Limami, Anis M.

    2017-04-01

    In cropping systems, legumes release substantial amounts of nitrogen (N) into the soil, via rhizodeposition, and constitute a sustainable source of N, instead of synthetic N fertilisers (Fustec et al. 2010). More frequent or/and intense droughts and floodings, due to climate change and intensification of agriculture, may affect N rhizodeposition (Preece & Peñuelas 2016). However, the effects of water stress on this process are poorly documented. A part of N derived from root exudates, mainly in amino acids (AAs) form, is suspected shape and regulate rhizosphere microbial community, thus playing a potential role in maintaining plant health in case of abiotic stress (Moe 2013). We hypothesized that root AA exudation could change significantly, according to water availability, and would help to understand N metabolism changes in plant-rhizosphere interactions. Because studying exudation from plant grown in unsterilized soil is challenging (Oburger et al. 2013), we have measured the rhizosphere AA fingerprint (RAAF), as the result of interactions between AA exudation and rhizospheric environment. In addition, plants were stem-labeled (cotton-wick) with 15N-urea for 72 h to provide direct evidence of a link between root AA and exudation in the soil. The RAAF was measured in Pisum sativum rhizosphere, under either a water deficit or a water excess for 72 h. Water deficit decreases biomass accumulation in shoots but not in roots. Then, water deficit had no significant effect on total AAs released into the rhizosphere but, it significantly modified the composition of RAAF, with a preferential increase of proline, alanine and glutamate and a rise in isotopic enrichment of AAs derived from oxaloacetate in tricarboxylic acidic cycle (asparagine, aspartate, threonine and isoleucine). These results support the idea that, under the early stages of water deficit, recently assimilated N is rapidly translocated to the roots, and part of it is exudated in AAs. Most of the exudated

  5. Isoenzymes of superoxide dismutase in nodules of Phaseolus vulgaris L. , Pisum sativum L. , and Vigna unguiculata (L. ) Walp

    SciTech Connect

    Becana, M.; Paris, F.J.; Sandalio, L.M.; Del Rio, L.A. Unidad de Bioquimica Vegetal, Granada )

    1989-08-01

    The activity and isozymic composition of superoxide dismutase were determined in nodules of Phaseolus vulgaris L., Pisum sativum L., and Vigna unguiculata (L.) Walp. A Mn-SOD was present in Rhizobium and two in Bradyrhizobium and bacteroids. Nodule mitochondria from all three legume species had a single Mn-SOD with similar relative mobility, whereas the cytosol contained several CuZn-SODs: two in Phaseolus and Pisum, and four in Vigna. In the cytoplasm of V. unguiculata nodules, a Fe-containing SOD was also present, with an electrophoretic mobility between those of CuZn- and Mn-SODs, and an estimated molecular weight of 57,000. Total SOD activity of the soluble fraction of host cells, expressed on a nodule fresh weight basis, exceeded markedly that of bacteroids. Likewise, specific SOD activities of free-living bacteria were superior or equal to those of their symbiotic forms. Soluble extracts of bacteria and bacteroids did not show peroxidase activity, but the nodule cell cytoplasm contained diverse peroxidase isozymes which were readily distinguishable from leghemoglobin components by electrophoresis. Data indicated that peroxidases and leghemoglobins did not significantly interfere with SOD localization on gels. Treatment with chloroform-ethanol scarcely affected the isozymic pattern of SODs and peroxidases, and had limited success in the removal of leghemoglobin.

  6. Different patterns of vein loading of exogenous ( sup 14 C)sucrose in leaves of pisum sativum and coleus blumei

    SciTech Connect

    Turgeon, R.; Wimmers, L.E. )

    1988-05-01

    Vein loading of exogenous ({sup 14}C)sucrose was studied using short uptake and wash periods to distinguish between direct loading into veins and loading via mesophyll tissue. Mature leaf tissue of Pisum sativum L. cv Little Marvel, or Coleus blumei Benth. cv Candidum, was abraded and leaf discs were floated on ({sup 14}C)sucrose solution for 1 or 2 minutes. Discs were then washed for 1 to 30 min either at room temperature or in the cold and were frozen, lyophilized, and autoradiographed. In P. sativum, veins were clearly labeled after 1 minute uptake and 1 minute wash periods. Autoradiographic images did not change appreciably with longer times of uptake or wash. Vein loading was inhibited by p-chloromercuribenzenesulfonic acid. These results indicate that uptake of exogenous sucrose occurs directly into the veins in this species. When C. blumei leaf discs were floated on ({sup 14}C)sucrose for 2 minutes and washed in the cold, the mesophyll was labeled but little, if any, minor vein loading occurred. When discs were labeled for 2 minutes and washed at room temperature, label was transferred from the mesophyll to the veins within minutes. These results indicate that there may be different patterns of phloem loading of photosynthetically derived sucrose in these two species.

  7. Alkali-Soluble Pectin Is the Primary Target of Aluminum Immobilization in Root Border Cells of Pea (Pisum sativum)

    PubMed Central

    Yang, Jin; Qu, Mei; Fang, Jing; Shen, Ren Fang; Feng, Ying Ming; Liu, Jia You; Bian, Jian Feng; Wu, Li Shu; He, Yong Ming; Yu, Min

    2016-01-01

    We investigated the hypothesis that a discrepancy of Al binding in cell wall constituents determines Al mobility in root border cells (RBCs) of pea (Pisum sativum), which provides protection for RBCs and root apices under Al toxicity. Plants of pea (P. sativum L. ‘Zhongwan no. 6’) were subjected to Al treatments under mist culture. The concentration of Al in RBCs was much higher than that in the root apex. The Al content in RBCs surrounding one root apex (104 RBCs) was approximately 24.5% of the total Al in the root apex (0–2.5 mm), indicating a shielding role of RBCs for the root apex under Al toxicity. Cell wall analysis showed that Al accumulated predominantly in alkali-soluble pectin (pectin 2) of RBCs. This could be attributed to a significant increase of uronic acids under Al toxicity, higher capacity of Al adsorption in pectin 2 [5.3-fold higher than that of chelate-soluble pectin (pectin 1)], and lower ratio of Al desorption from pectin 2 (8.5%) compared with pectin 1 (68.5%). These results indicate that pectin 2 is the primary target of Al immobilization in RBCs of pea, which impairs Al access to the intracellular space of RBCs and mobility to root apices, and therefore protects root apices and RBCs from Al toxicity. PMID:27679639

  8. Proteomic Profiling of the Microsomal Root Fraction: Discrimination of Pisum sativum L. Cultivars and Identification of Putative Root Growth Markers

    PubMed Central

    Meisrimler, Claudia-Nicole; Wienkoop, Stefanie; Lüthje, Sabine

    2017-01-01

    Legumes are a large and economically important family, containing a variety of crop plants. Alongside different cereals, some fruits, and tropical roots, a number of leguminosae evolved for millennia as crops with human society. One of these legumes is Pisum sativum L., the common garden pea. In the past, breeding has been largely selective on improved above-ground organs. However, parameters, such as root-growth, which determines acquisition of nutrients and water, have largely been underestimated. Although the genome of P. sativum is still not fully sequenced, multiple proteomic studies have been published on a variety of physiological aspects in the last years. The presented work focused on the connection between root length and the influence of the microsomal root proteome of four different pea cultivars after five days of germination (cultivar Vroege, Girl from the Rhineland, Kelvedon Wonder, and Blauwschokker). In total, 60 proteins were identified to have significantly differential abundances in the four cultivars. Root growth of five-days old seedlings and their microsomal proteome revealed a similar separation pattern, suggesting that cultivar-specific root growth performance is explained by differential membrane and ribosomal protein levels. Hence, we reveal and discuss several putative root growth protein markers possibly playing a key role for improved primary root growth breeding strategies. PMID:28257117

  9. Proteomic Profiling of the Microsomal Root Fraction: Discrimination of Pisum sativum L. Cultivars and Identification of Putative Root Growth Markers.

    PubMed

    Meisrimler, Claudia-Nicole; Wienkoop, Stefanie; Lüthje, Sabine

    2017-03-02

    Legumes are a large and economically important family, containing a variety of crop plants. Alongside different cereals, some fruits, and tropical roots, a number of leguminosae evolved for millennia as crops with human society. One of these legumes is Pisum sativum L., the common garden pea. In the past, breeding has been largely selective on improved above-ground organs. However, parameters, such as root-growth, which determines acquisition of nutrients and water, have largely been underestimated. Although the genome of P. sativum is still not fully sequenced, multiple proteomic studies have been published on a variety of physiological aspects in the last years. The presented work focused on the connection between root length and the influence of the microsomal root proteome of four different pea cultivars after five days of germination (cultivar Vroege, Girl from the Rhineland, Kelvedon Wonder, and Blauwschokker). In total, 60 proteins were identified to have significantly differential abundances in the four cultivars. Root growth of five-days old seedlings and their microsomal proteome revealed a similar separation pattern, suggesting that cultivar-specific root growth performance is explained by differential membrane and ribosomal protein levels. Hence, we reveal and discuss several putative root growth protein markers possibly playing a key role for improved primary root growth breeding strategies.

  10. Functional characterization of PeIF5B as eIF5B homologue from Pisum sativum.

    PubMed

    Rasheedi, Sheeba; Suragani, Madhuri; Raviprasad, Podili; Ghosh, Sudip; Suragani, Rajasekhar N V S; Ramaiah, Kolluru V A; Ehtesham, Nasreen Z

    2015-11-01

    We earlier reported 'PeIF5B' as a novel factor from Pisum sativum that has sequence similarity to eIF5B (S. Rasheedi, S. Ghosh, M. Suragani et al., P. sativum contains a factor with strong homology to eIF5B, Gene 399 (2007) 144-151). The main aim of the present study was to perform functional characterization of PeIF5B as an eIF5B homologue from plant system. PeIF5B shows binding to Met - tRNA(f)(Met), hydrolyses GTP and interacts with ribosomes. In vivo growth complementation analysis shows that PeIF5B partially complements its yeast homologue. Interestingly, PeIF5B mainly localizes in the nucleus as confirmed by nuclear localization signal (NLS) prediction, confocal imaging and immunoblots of cellular fractions. Similar to the yeast eIF5B but unlike the human orthologue, PeIF5B is an intron-less gene. This study highlights PeIF5B's role as a functional eIF5B homologue possibly participating in nuclear translation in plant system. Copyright © 2015 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  11. Alkali-Soluble Pectin Is the Primary Target of Aluminum Immobilization in Root Border Cells of Pea (Pisum sativum).

    PubMed

    Yang, Jin; Qu, Mei; Fang, Jing; Shen, Ren Fang; Feng, Ying Ming; Liu, Jia You; Bian, Jian Feng; Wu, Li Shu; He, Yong Ming; Yu, Min

    2016-01-01

    We investigated the hypothesis that a discrepancy of Al binding in cell wall constituents determines Al mobility in root border cells (RBCs) of pea (Pisum sativum), which provides protection for RBCs and root apices under Al toxicity. Plants of pea (P. sativum L. 'Zhongwan no. 6') were subjected to Al treatments under mist culture. The concentration of Al in RBCs was much higher than that in the root apex. The Al content in RBCs surrounding one root apex (10(4) RBCs) was approximately 24.5% of the total Al in the root apex (0-2.5 mm), indicating a shielding role of RBCs for the root apex under Al toxicity. Cell wall analysis showed that Al accumulated predominantly in alkali-soluble pectin (pectin 2) of RBCs. This could be attributed to a significant increase of uronic acids under Al toxicity, higher capacity of Al adsorption in pectin 2 [5.3-fold higher than that of chelate-soluble pectin (pectin 1)], and lower ratio of Al desorption from pectin 2 (8.5%) compared with pectin 1 (68.5%). These results indicate that pectin 2 is the primary target of Al immobilization in RBCs of pea, which impairs Al access to the intracellular space of RBCs and mobility to root apices, and therefore protects root apices and RBCs from Al toxicity.

  12. Immunolocalization of dually phosphorylated MAPKs in dividing root meristem cells of Vicia faba, Pisum sativum, Lupinus luteus and Lycopersicon esculentum.

    PubMed

    Winnicki, Konrad; Żabka, Aneta; Bernasińska, Joanna; Matczak, Karolina; Maszewski, Janusz

    2015-06-01

    In plants, phosphorylated MAPKs display constitutive nuclear localization; however, not all studied plant species show co-localization of activated MAPKs to mitotic microtubules. The mitogen-activated protein kinase (MAPK) signaling pathway is involved not only in the cellular response to biotic and abiotic stress but also in the regulation of cell cycle and plant development. The role of MAPKs in the formation of a mitotic spindle has been widely studied and the MAPK signaling pathway was found to be indispensable for the unperturbed course of cell division. Here we show cellular localization of activated MAPKs (dually phosphorylated at their TXY motifs) in both interphase and mitotic root meristem cells of Lupinus luteus, Pisum sativum, Vicia faba (Fabaceae) and Lycopersicon esculentum (Solanaceae). Nuclear localization of activated MAPKs has been found in all species. Co-localization of these kinases to mitotic microtubules was most evident in L. esculentum, while only about 50% of mitotic cells in the root meristems of P. sativum and V. faba displayed activated MAPKs localized to microtubules during mitosis. Unexpectedly, no evident immunofluorescence signals at spindle microtubules and phragmoplast were noted in L. luteus. Considering immunocytochemical analyses and studies on the impact of FR180204 (an inhibitor of animal ERK1/2) on mitotic cells, we hypothesize that MAPKs may not play prominent role in the regulation of microtubule dynamics in all plant species.

  13. Rates of Sugar Uptake by Guard Cell Protoplasts of Pisum sativum L. Related to the Solute Requirement for Stomatal Opening1

    PubMed Central

    Ritte, Gerhard; Rosenfeld, Johanna; Rohrig, Kerstin; Raschke, Klaus

    1999-01-01

    We wished to determine whether the capacity of the sugar uptake mechanisms of guard cells of the Argenteum mutant of pea (Pisum sativum L.) sufficed to support a concurrent stomatal opening movement. Sugar uptake by guard cell protoplasts was determined by silicone-oil-filtering centrifugation. The protoplasts took up [14C]glucose, [14C]fructose, and [14C]sucrose (Suc), apparently in symport with protons. Mannose, galactose, and fructose competed with Glc for transport by a presumed hexose carrier. The uptake of Glc saturated with a Km of 0.12 mm and a Vmax of 19 fmol cell−1 h−1. At external concentrations <1 mm, the uptake of Suc was slower than that of Glc. It exhibited a saturating component with a Km varying between 0.25 and 0.8 mm and a Vmax between 1 and 10 fmol cell−1 h−1, and at external concentrations >1 mm, a non-saturating component. At apoplastic sugar concentrations below 4 mm, sugar import was estimated to be mainly in the form of hexoses and too slow to support a simultaneous stomatal opening movement. If, however, during times of high photosynthesis and transpiration, the apoplastic Suc concentration rose and entered the range of non-saturating import, absorbed Suc could replace potassium malate as the osmoticum for the maintenance of stomatal opening. PMID:10517857

  14. Arginine Decarboxylase and Putrescine Oxidase in Ovaries of Pisum sativum L. (Changes during Ovary Senescence and Early Stages of Fruit Development).

    PubMed Central

    Perez-Amador, M. A.; Carbonell, J.

    1995-01-01

    Enzymatic activities involved in putrescine metabolism in ovaries of Pisum sativum L. during ovary senescence and fruit set were investigated. Accumulation of putrescine was observed during incubation of extracts from gibberellic acid-treated unpollinated ovaries (young developing fruits) but not in extracts from untreated ovaries (senescent ovaries). Extracts from pea ovaries showed arginine decarboxylase (ADC) activity, but ornithine decarboxylase and arginase activity were not detected. ADC activity decreased in presenescent ovaries and increased markedly after induction of fruit set with gibberellic acid. Increases in ADC activity were also observed with application of other plant growth substances (benzy-ladenine and 2,4-dichlorophenoxyacetic acid), after pollination, and in the slender (la crys) pea mutant. By contrast, putrescine oxidase activity increased in presenescent ovaries but did not increase during early fruit development. All of these results suggest that ADC and putrescine oxidase are involved in the control of putrescine metabolism. Ovary senescence is characterized by the absence of putrescine biosynthesis enzymes and increased levels of putrescine oxidase and fruit development by an increase in ADC and a constant level of putrescine oxidase. PMID:12228409

  15. Signal Integration by ABA in the Blue Light-Induced Acidification of Leaf Pavement Cells in Pea (Pisum sativum L. var. Argenteum)

    PubMed Central

    den Os, Désirée; Staal, Marten

    2007-01-01

    Leaf pavement cell expansion in light depends on apoplastic acidification by a plasma membrane proton-pumping ATPase, modifying cell wall extensibility and providing the driving force for uptake of osmotically active solutes generating turgor. This paper shows that the plant hormone ABA inhibits light-induced leaf disk growth as well as the blue light-induced pavement cell growth in pea (Pisum sativum L.). In the phytochrome chromophore-deficient mutant pcd2, the effect of ABA on the blue light-induced apoplastic acidification response, which exhibits a high fluence phase via phytochrome and a low fluence phase via an unknown blue light receptor, is still present, indicating an interaction of ABA with the blue light receptor pathway. Furthermore, it is shown that ABA inhibits the blue light-induced apoplastic acidification reversibly. These results indicate that the effect of ABA on apoplastic acidification can provide a mechanism for short term, reversible adjustment of leaf growth rate to environmental change. PMID:19516983

  16. Recessive Resistance in Pisum sativum and Potyvirus Pathotype Resolved in a Gene-for-Cistron Correspondence between Host and Virus

    PubMed Central

    Johansen, I. Elisabeth; Lund, Ole Søgaard; Hjulsager, Charlotte K.; Laursen, Jesper

    2001-01-01

    Pea seed-borne mosaic potyvirus (PSbMV) isolates are divided into pathotypes P-1, P-2, and P-4 according to their infection profile on a panel of Pisum sativum lines. P. sativum PI 269818 is resistant to P-1 and P-2 isolates and is susceptible to P-4 isolates. Resistance to P-1 is inherited as a single recessive gene, denoted sbm-1, and the pathogenicity determinant has previously been mapped to the virus-coded protein VPg. In the cultivar Bonneville, a second recessive gene, sbm-2, confers specific resistance to P-2. By exchanging cistrons between a P-2 and a P-4 isolate, the P3-6k1 cistron was identified as the PSbMV host-specific pathogenicity determinant on Bonneville. Exchange of P3-6k1 did not affect infection on PI 269818, and infection of Bonneville was not altered by substitution of the VPg cistron, indicating that P3-6k1 and VPg are independent determinants of pathotype-specific infectivity. On PI 269818 the pathogenicity determinant of both P-1 and P-2 mapped to the N terminus of VPg. This suggests that VPg from the P-1 and P-2 isolates are functionally similar on this host and that resistance to P-1 and P-2 in PI 269818 may operate by the same mechanism. Identification of VPg–sbm-1 and P3-6k1–sbm-2 as independent pairs of genetic interactors between PSbMV and P. sativum provides a simple explanation of the three known pathotypes of PSbMV. Furthermore, analysis of β-glucuronidase-tagged P-2 virus indicated that sbm-2 resistance affected an early step in infection, implying that the P3-6k1 region plays a critical role in potyvirus replication or cell-to-cell movement. PMID:11413328

  17. High-quality permanent draft genome sequence of Rhizobium leguminosarum bv. viciae strain GB30; an effective microsymbiont of Pisum sativum growing in Poland

    SciTech Connect

    Mazur, Andrzej; De Meyer, Sofie E.; Tian, Rui; Wielbo, Jerzy; Zebracki, Kamil; Seshadri, Rekha; Reddy, T. B.K.; Markowitz, Victor; Ivanova, Natalia N.; Pati, Amrita; Woyke, Tanja; Kyrpides, Nikos C.; Reeve, Wayne

    2015-07-16

    We report that Rhizobium leguminosarum bv. viciae GB30 is an aerobic, motile, Gram-negative, non-spore-forming rod that can exist as a soil saprophyte or as a legume microsymbiont of Pisum sativum. GB30 was isolated in Poland from a nodule recovered from the roots of Pisum sativum growing at Janow. GB30 is also an effective microsymbiont of the annual forage legumes vetch and pea. Here we describe the features of R. leguminosarum bv. viciae strain GB30, together with sequence and annotation. The 7,468,464 bp high-quality permanent draft genome is arranged in 78 scaffolds of 78 contigs containing 7,227 protein-coding genes and 75 RNA-only encoding genes, and is part of the GEBA-RNB project proposal.

  18. High-quality permanent draft genome sequence of Rhizobium leguminosarum bv. viciae strain GB30; an effective microsymbiont of Pisum sativum growing in Poland

    DOE PAGES

    Mazur, Andrzej; De Meyer, Sofie E.; Tian, Rui; ...

    2015-07-16

    We report that Rhizobium leguminosarum bv. viciae GB30 is an aerobic, motile, Gram-negative, non-spore-forming rod that can exist as a soil saprophyte or as a legume microsymbiont of Pisum sativum. GB30 was isolated in Poland from a nodule recovered from the roots of Pisum sativum growing at Janow. GB30 is also an effective microsymbiont of the annual forage legumes vetch and pea. Here we describe the features of R. leguminosarum bv. viciae strain GB30, together with sequence and annotation. The 7,468,464 bp high-quality permanent draft genome is arranged in 78 scaffolds of 78 contigs containing 7,227 protein-coding genes and 75more » RNA-only encoding genes, and is part of the GEBA-RNB project proposal.« less

  19. High-quality permanent draft genome sequence of Rhizobium leguminosarum bv. viciae strain GB30; an effective microsymbiont of Pisum sativum growing in Poland.

    PubMed

    Mazur, Andrzej; De Meyer, Sofie E; Tian, Rui; Wielbo, Jerzy; Zebracki, Kamil; Seshadri, Rekha; Reddy, Tbk; Markowitz, Victor; Ivanova, Natalia N; Pati, Amrita; Woyke, Tanja; Kyrpides, Nikos C; Reeve, Wayne

    2015-01-01

    Rhizobium leguminosarum bv. viciae GB30 is an aerobic, motile, Gram-negative, non-spore-forming rod that can exist as a soil saprophyte or as a legume microsymbiont of Pisum sativum. GB30 was isolated in Poland from a nodule recovered from the roots of Pisum sativum growing at Janow. GB30 is also an effective microsymbiont of the annual forage legumes vetch and pea. Here we describe the features of R. leguminosarum bv. viciae strain GB30, together with sequence and annotation. The 7,468,464 bp high-quality permanent draft genome is arranged in 78 scaffolds of 78 contigs containing 7,227 protein-coding genes and 75 RNA-only encoding genes, and is part of the GEBA-RNB project proposal.

  20. Draft Genome Sequence of the Plant Growth-Promoting Rhizobacterium Pseudomonas fluorescens Strain CREA-C16 Isolated from Pea (Pisum sativum L.) Rhizosphere

    PubMed Central

    Sorrentino, Roberto; Scotti, Riccardo; Salzano, Melania; Aurilia, Vincenzo

    2017-01-01

    ABSTRACT Herein, we report the draft genome sequence of Pseudomonas fluorescens strain CREA-C16, a plant growth-promoting rhizobacterium that was isolated from the rhizosphere of Pisum sativum L. plants. The genome sequence is ~6 Mb in size, with a G+C content of 60.1%, and includes 4,457 candidate protein-encoding genes. PMID:28126933

  1. Draft Genome Sequence of the Plant Growth-Promoting Rhizobacterium Pseudomonas fluorescens Strain CREA-C16 Isolated from Pea (Pisum sativum L.) Rhizosphere.

    PubMed

    D'Agostino, Nunzio; Sorrentino, Roberto; Scotti, Riccardo; Salzano, Melania; Aurilia, Vincenzo; Zaccardelli, Massimo

    2017-01-26

    Herein, we report the draft genome sequence of Pseudomonas fluorescens strain CREA-C16, a plant growth-promoting rhizobacterium that was isolated from the rhizosphere of Pisum sativum L. plants. The genome sequence is ~6 Mb in size, with a G+C content of 60.1%, and includes 4,457 candidate protein-encoding genes. Copyright © 2017 D’Agostino et al.

  2. Iron Transport to Developing Ovules of Pisum sativum (I. Seed Import Characteristics and Phloem Iron-Loading Capacity of Source Regions).

    PubMed Central

    Grusak, M. A.

    1994-01-01

    To understand the processes that control Fe transport to developing seeds, we have characterized seed growth and Fe accretion and have developed a radiotracer technique for quantifying phloem Fe loading in vegetative source regions of Pisum sativum. In hydroponically grown plants of cv Sparkle, developing ovules exhibited a seed-growth period of 22 d, with Fe import occurring throughout the 22-d period. Average Fe content of mature seeds was 19 [mu]g. Source tissues of intact plants were abraded and pulse labeled for 4 h with 100 [mu]M 59Fe(III)-citrate. Fe was successfully phloem loaded and transported to seeds from leaflets, stipules, and pod walls. Total export of 59Fe from labeled source regions was used to calculate tissue-loading rates of 36, 40, and 51 pmol of Fe cm-2 h-1 for the leaflet, stipule, and pod wall surfaces, respectively. By comparison, surface area measurements, along with seed-growth results, allowed us to calculate average theoretical influx values of 42 or 68 pmol of Fe cm-2 h-1 for vegetative tissues at nodes with one or two pods, respectively. Additional studies with the regulatory pea mutant, E107 (a single-gene mutant of cv Sparkle that can overaccumulate Fe), enabled us to increase Fe delivery endogenously to the vegetative tissues. A 36-fold increase in Fe content of E107 leaves, relative to Sparkle, resulted in no increase in Fe content of E107 seeds. Based on these findings, we hypothesized that Fe is phloem loaded in a chelated form, and the expression/synthesis of the endogenous chelator is an important factor in the control of Fe transport to the seeds. PMID:12232115

  3. Long-term iron deficiency: Tracing changes in the proteome of different pea (Pisum sativum L.) cultivars.

    PubMed

    Meisrimler, Claudia-Nicole; Wienkoop, Stefanie; Lyon, David; Geilfus, Christoph-Martin; Lüthje, Sabine

    2016-05-17

    Iron deficiency (-Fe) is one of the major problems in crop production. Dicots, like pea (Pisum sativum L.), are Strategy I plants, which induce a group of specific enzymes such as Fe(III)-chelate reductase (FRO), Fe responsive transporter (IRT) and H(+)-ATPase (HA) at the root plasma membrane under -Fe. Different species and cultivars have been shown to react diversely to -Fe. Furthermore, different kinds of experimental set-ups for -Fe have to be distinguished: i) short-term vs. long-term, ii) constant vs. acute alteration and iii) buffered vs. unbuffered systems. The presented work compares the effects of constant long-term -Fe in an unbuffered system on roots of four different pea cultivars in a timely manner (12, 19 and 25days). To differentiate the effects of -Fe and plant development, control plants (+Fe) were analyzed in comparison to -Fe plants. Besides physiological measurements, an integrative study was conducted using a comprehensive proteome analysis. Proteins, related to stress adaptation (e.g. HSP), reactive oxygen species related proteins and proteins of the mitochondrial electron transport were identified to be changed in their abundance. Regulations and possible functions of identified proteins are discussed. Pea (Pisum sativum L.) belongs to the legume family (Fabaceae) and is an important crop plant due to high Fe, starch and protein contents. According to FAOSTAT data (September 2015), world production of the garden pea quadrupled from 1970 to 2012. Since the initial studies by Gregor Mendel, the garden pea became the most-characterized legume and has been used in numerous investigations in plant biochemistry and physiology, but is not well represented in the "omics"-related fields. A major limitation in pea production is the Fe availability from soils. Adaption mechanisms to Fe deficiency vary between species, and even cultivars have been shown to react diversely. A label-free proteomic approach, in combination with physiological measurements

  4. Enhancing Neoplasm Expression in Field Pea (Pisum sativum) via Intercropping and Its Significance to Pea Weevil (Bruchus pisorum) Management

    PubMed Central

    Teshome, Abel; Bryngelsson, Tomas; Mendesil, Esayas; Marttila, Salla; Geleta, Mulatu

    2016-01-01

    Neoplasm formation, a non-meristematic tissue growth on young field pea (Pisum sativum L.) pods is triggered in the absence of UV light and/or in response to oviposition by pea weevil (Bruchus pisorum L.). This trait is expressed in some genotypes [neoplastic (Np) genotypes] of P. sativum and has the capacity to obstruct pea weevil larval entry into developing seeds. In the present study, 26% of the tested accessions depicted the trait when grown under greenhouse conditions. However, UV light inhibits full expression of this trait and subsequently it is inconspicuous at the field level. In order to investigate UV light impact on the expression of neoplasm, particular Np genotypes were subjected to UV lamp light exposure in the greenhouse and sunlight at the field level. Under these different growing conditions, the highest mean percentage of Np pods was in the control chamber in the greenhouse (36%) whereas in single and double UV lamp chambers, the percentage dropped to 10 and 15%, respectively. Furthermore, when the same Np genotypes were grown in the field, the percentage of Np pods dropped significantly (7%). In order to enhance Np expression at the field level, intercropping of Np genotypes with sorghum was investigated. As result, the percentage of Np pods was threefold in intercropped Np genotypes as compared to those without intercropping. Therefore, intercropping Np genotypes with other crops such as sorghum and maize can facilitate neoplasm formation, which in turn can minimize the success rate of pea weevil larvae entry into developing seeds. Greenhouse artificial infestation experiments showed that pea weevil damage in Np genotypes is lower in comparison to wild type genotypes. Therefore, promoting Np formation under field conditions via intercropping can serve as part of an integrated pea weevil management strategy especially for small scale farming systems. PMID:27242855

  5. Hypoxic stress triggers a programmed cell death pathway to induce vascular cavity formation in Pisum sativum roots.

    PubMed

    Sarkar, Purbasha; Gladish, Daniel K

    2012-12-01

    Flooding at warm temperatures induces hypoxic stress in Pisum sativum seedling roots. In response, some undifferentiated cells in the primary root vascular cylinder start degenerating and form a longitudinal vascular cavity. Changes in cellular morphology and cell wall ultrastructure detected previously in the late stages of cavity formation suggest possible involvement of programmed cell death (PCD). In this study, cytological events occurring in the early stages of cavity formation were investigated. Systematic DNA fragmentation, a feature of many PCD pathways, was detected in the cavity-forming roots after 3 h of flooding in situ by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay and in isolated total DNA by gel electrophoresis. High molecular weight DNA fragments of about 20-30 kb were detected by pulse-field gel electrophoresis, but no low-molecular weight internucleosomal DNA fragments were detected by conventional gel electrophoresis. Release of mitochondrial cytochrome c protein into the cytosol, an integral part of mitochondria-dependent PCD pathways, was detected in the cavity-forming roots within 2 h of flooding by fluorescence microscopy of immunolabeled cytochrome c in situ and in isolated mitochondrial and cytosolic protein fractions by western blotting. DNA fragmentation and cytochrome c release remained confined to the undifferentiated cells in center of the root vascular cylinders, even after 24 h of flooding, while outer vascular cylinder cells and cortical cells maintained cellular integrity and normal activity. These findings confirm that hypoxia-induced vascular cavity formation in P. sativum roots involves PCD, and provides a chronological model of cytological events involved in this rare and understudied PCD system.

  6. Enhancing Neoplasm Expression in Field Pea (Pisum sativum) via Intercropping and Its Significance to Pea Weevil (Bruchus pisorum) Management.

    PubMed

    Teshome, Abel; Bryngelsson, Tomas; Mendesil, Esayas; Marttila, Salla; Geleta, Mulatu

    2016-01-01

    Neoplasm formation, a non-meristematic tissue growth on young field pea (Pisum sativum L.) pods is triggered in the absence of UV light and/or in response to oviposition by pea weevil (Bruchus pisorum L.). This trait is expressed in some genotypes [neoplastic (Np) genotypes] of P. sativum and has the capacity to obstruct pea weevil larval entry into developing seeds. In the present study, 26% of the tested accessions depicted the trait when grown under greenhouse conditions. However, UV light inhibits full expression of this trait and subsequently it is inconspicuous at the field level. In order to investigate UV light impact on the expression of neoplasm, particular Np genotypes were subjected to UV lamp light exposure in the greenhouse and sunlight at the field level. Under these different growing conditions, the highest mean percentage of Np pods was in the control chamber in the greenhouse (36%) whereas in single and double UV lamp chambers, the percentage dropped to 10 and 15%, respectively. Furthermore, when the same Np genotypes were grown in the field, the percentage of Np pods dropped significantly (7%). In order to enhance Np expression at the field level, intercropping of Np genotypes with sorghum was investigated. As result, the percentage of Np pods was threefold in intercropped Np genotypes as compared to those without intercropping. Therefore, intercropping Np genotypes with other crops such as sorghum and maize can facilitate neoplasm formation, which in turn can minimize the success rate of pea weevil larvae entry into developing seeds. Greenhouse artificial infestation experiments showed that pea weevil damage in Np genotypes is lower in comparison to wild type genotypes. Therefore, promoting Np formation under field conditions via intercropping can serve as part of an integrated pea weevil management strategy especially for small scale farming systems.

  7. Foliar Application of Plant Growth-Promoting Rhizobacteria Increases Antifungal Compounds in Pea (Pisum sativum) Against Erysiphe pisi.

    PubMed

    Bahadur, A; Singh, U P; Sarma, B K; Singh, D P; Singh, K P; Singh, A

    2007-09-01

    Systemic effect of two plant growth-promoting rhizobacterial (PGPR) strains,viz., Pseudomonas fluorescens (Pf4) and P. aeruginosa (Pag), was evaluated on pea (Pisum sativum) against the powdery mildew pathogen Erysiphe pisi. Foliar spray of the two PGPR strains was done on specific nodal leaves of pea and conidial germination of E. pisi was observed on other nodal leaves,distal to the treated ones. Conidial germination was reduced on distant leaves and at the same time,specific as well as total phenolic compounds increased in the leaves distal to those applied with PGPR strains,thereby indicating a positive correlation. The strains induced accumulation of phenolic compounds in pea leaves and the amount increased when such leaves were get inoculated with E. pisi conidia. Between the two strains, Pag was found to be more effective than Pf4 as its effect was more persistent in pea leaves. Foliar application of PGPR strains for the control of powdery mildew of pea is demonstrated in vitro while correlating it with the increased accumulation of plant phenolics.

  8. Novel plant-microbe rhizosphere interaction involving Streptomyces lydicus WYEC108 and the pea plant (Pisum sativum).

    PubMed

    Tokala, Ranjeet K; Strap, Janice L; Jung, Carina M; Crawford, Don L; Salove, Michelle Hamby; Deobald, Lee A; Bailey, J Franklin; Morra, M J

    2002-05-01

    A previously undescribed plant-microbe interaction between a root-colonizing Streptomyces species, S. lydicus WYEC108, and the legume Pisum sativum is described. The interaction is potentially of great importance to the health and growth in nature of this nodulating legume. The root-colonizing soil actinomycete S. lydicus WYEC108 influences pea root nodulation by increasing root nodulation frequency, possibly at the level of infection by Rhizobium spp. S. lydicus also colonizes and then sporulates within the surface cell layers of the nodules. Colonization leads to an increase in the average size of the nodules that form and improves the vigor of bacteroids within the nodules by enhancing nodular assimilation of iron and possibly other soil nutrients. Bacteroid accumulation of the carbon storage polymer, poly-beta-hydroxybutyrate, is reduced in colonized nodules. Root nodules of peas taken from agricultural fields in the Palouse hills of northern Idaho were also found to be colonized by actinomycete hyphae. We hypothesize that root and nodule colonization is one of several mechanisms by which Streptomyces acts as a naturally occurring plant growth-promoting bacterium in pea and possibly other leguminous plants.

  9. Symbiotic dinitrogen fixation as affected by short-term application of nitrate to nodulated Pisum sativum L.

    PubMed

    Skrdleta, V; Gaudinová, A; Nĕmcová, M; Hyndráková, A

    1980-01-01

    Effect of nitrate on the nitrogenase (C2H2-reduction) activity, growth of nodule tissue accumulation of nitrate and nitrate reductase activity in 4-weeks-old nodulated peas (Pisum sativum L.) was investigated. A relatively slow decrease of the total nitrogenase activity (mumol C2H4 per root per h), as compared with plants cultivated without nitrate, was due to both retardation of further growth of the nodule tissue and to a decrease of their specific nitrogenase activity (mumol C2H4 per gf.wt. per h). However, an absolute and pronounced decrease of both nitrogenase activities occurred only 4 or 7 d after the application of nitrate. The addition of nitrate led to its rapid accumulation in the nodule and leaf tissue with a simultaneous induction of the nitrate reductase activity. The nitrogenase activity was not completely inhibited even after a 7-d cultivation with 280 m NO3- -N in the nutrient medium and after accumulation of up to 180 ppm NO3- -Nf.wt. in the nodule tissue. The results obtained indicate that the "photosynthate deprivation" reflects competition between assimilation of nitrate and fixation of dinitrogen.

  10. [Research progress on the cloning of Mendel's gene in pea (Pisum sativum L.) and its application in genetics teaching].

    PubMed

    He, Feng-Hua; Zhu, Bi-Yan; Gao, Feng; Li, Shao-Shan; Li, Niang-Hui

    2013-07-01

    One hundred and fifty years ago, Gregor Mendel investigated the segregation of seven traits in pea (Pisum sativum) and established the law of segregation and the law of independent assortment in genetics. After the two laws of genetics were rediscovered in 1900, the seven traits have been extensively investigated in the fields of plant physiology and biochemistry as well as in the cell and molecular levels. Recently, with the development of molecular technology in genetics, four genes for seed shape (R), stem length (Le), cotyledon colour (I), and flower colour (A) have been cloned and sequenced; and another three genes for immature pod colour (Gp), fasciation (Fa) and pod form (V) have been located in the linkage groups, respectively. The identification and cloning of the four Mendel's genes will help deeply understand the basic concept of gene in many respects: like the diversity of gene function, the different origins for gene mutation in molecular level, and the molecular nature of a dominant gene or a recessive gene. In teaching of genetics, the introduction of most recent research advancements of cloning of Mendel's genes to the students and the interpretation of the Mendel's laws in molecular level will help students promote their learning interests in genetics and help students grasp the whole content from classical genetics to molecular genetics and the developmental direction of this subject.

  11. A proposed interplay between peroxidase, amine oxidase and lipoxygenase in the wounding-induced oxidative burst in Pisum sativum seedlings.

    PubMed

    Roach, Thomas; Colville, Louise; Beckett, Richard P; Minibayeva, Farida V; Havaux, Michel; Kranner, Ilse

    2015-04-01

    Plant surfaces form the barrier between a plant and its environment. Upon damage, the wound healing process begins immediately and is accompanied by a rapid production of extracellular reactive oxygen species (ROS), essential in deterring pathogens, signalling responses and cell wall restructuring. Although many enzymes produce extracellular ROS, it is unclear if ROS-producing enzymes act synergistically. We characterised the oxidative burst of superoxide (O2(·-)) and hydrogen peroxide (H2O2) that follows wounding in pea (Pisum sativum L.) seedlings. Rates of ROS production were manipulated by exogenous application of enzyme substrates and inhibitors. The results indicate significant roles for di-amine oxidases (DAO) and peroxidases (Prx) rather than NADPH oxidase. The burst of O2(·-) was strongly dependent on the presence of H2O2 produced by DAO. Potential substrates released from wounded seedlings included linoleic acid that, upon exogenous application, strongly stimulated catalase-sensitive O2(·-) production. Moreover, a 65kD plasma membrane (PM) guaiacol Prx was found in the secretome of wounded seedlings and showed dependence on linoleic acid for O2(·-) production. Lipoxygenases are suggested to modulate O2(·-) production by consuming polyunsaturated fatty acids in the apoplast. Overall, a O2(·-)-producing mechanism involving H2O2-derived from DAO, linoleic acid and a PM-associated Prx is proposed. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Root and nodule growth in Pisum sativum L. in relation to photosynthesis: analysis using 13C-labelling.

    PubMed

    Voisin, A S; Salon, C; Jeudy, C; Warembourg, F R

    2003-10-01

    The effect of the nitrogen source (gaseous nitrogen, N2, or nitrate ions, NO3-) on the use of carbon (C) for root and nodule growth of pea (Pisum sativum L.) was investigated using 13C-labelling of assimilated CO2 at various stages of growth. Nitrate supply and growing conditions (sowing dates, air CO2 concentration) were varied to alter photosynthetic rates. Nodules are the sink with the highest demand for C in both the vegetative and flowering stages, growing at the expense of shoot and root in the vegetative stage, but only at the expense of roots at flowering. Until flowering, the addition of C into root and nodule biomass was linearly related to pre-existing biomass, thus determining net sink strengths which decreased with root and nodule age. Nodule growth patterns did not depend on the N source, whereas root growth was increased by nitrate when nodule biomass was low. At seed filling, the increase in C of biomass of the root system was no longer related to pre-existing biomass and C was preferentially diverted to roots of plants assimilating nitrate, or to nodules for plants fixing N2.

  13. The loss of carbon-20 in C19-gibberellin biosynthesis in a cell-free system from Pisum sativum L.

    PubMed

    Kamiya, Y; Takahashi, N; Graebe, J E

    1986-12-01

    The fate of the carbon-20 atom in gibberellin (GA) biosynthesis was studied in a cell-free system from Pisum sativum. This carbon atom is lost at the aldehyde stage of oxidation when C20-GAs are converted to C19-GAs. Gibberellin A12 labeled with (14)C at C-20 was prepared from [3'-(14)C]mevalonic acid with a cell-free system from Cucurbita maxima and incubated with the pea system. Analysis of the gas and aqueous phases showed that (14)CO2 was formed at the same rate and in nearly equivalent amounts as (14)C-labeled C19-GAs whereas [(14)C]formic acid and [(14)C]formaldehyde were not detectable. The possibility that C-20 had been lost as formic acid which had then been converted to CO2 was investigated by control incubations with [(14)C]formic acid. The rate of release of (14)CO2 from [(14)C]formic acid was only one fiftieth of the rate of (14)CO2 release from [(14)C]GA12 as the substrate. We conclude that in the formation of C19-GAs from C20-GAs, the C-20 is removed directly as CO2.

  14. Net Photosynthesis, Electron Transport Capacity, and Ultrastructure of Pisum sativum L. Exposed to Ultraviolet-B Radiation 1

    PubMed Central

    Brandle, James R.; Campbell, William F.; Sisson, William B.; Caldwell, Martyn M.

    1977-01-01

    Pisum sativum L. was exposed to ultraviolet-B (UV-B) radiation (280-315 nm) in greenhouse and controlled environment chambers to examine the effect of this radiation on photosynthetic processes. Net photosynthetic rates of intact leaves were reduced by UV-B irradiation. Stable leaf diffusion resistances indicated that the impairment of photosynthesis did not involve the simple limitation of CO2 diffusion into the leaf. Dark respiration rates were increased by previous exposure to this radiation. Electron transport capacity as indicated by methylviologen reduction was also sensitive to UV-B irradiation. The ability of ascorbate-reduced 2,6-dichlorophenolindophenol to restore much of the electron transport capacity of the UV-B-irradiated plant material suggested that inhibition by this radiation was more closely associated with photosystem II than with photosystem I. Electron micrographs indicated structural damage to chloroplasts as well as other organelles. Plant tissue irradiated for only 15 minutes exhibited dilation of thylakoid membranes of the chloroplast in some cells. Some reduction in Hill reaction activity was also evidenced in these plant materials which had been irradiated for periods as short as 15 minutes. Images PMID:16660029

  15. Differential Gene Expression in the Meristem and during Early Fruit Growth of Pisum sativum L. Identifies Potential Targets for Breeding

    PubMed Central

    Smitha Ninan, Annu; Shah, Anish; Song, Jiancheng; Jameson, Paula E.

    2017-01-01

    For successful molecular breeding it is important to identify targets to the gene family level, and in the specific species of interest, in this case Pisum sativum L. The cytokinins have been identified as a key breeding target due to their influence on plant architecture, and on seed size and sink activity. We focused on the cytokinin biosynthetic gene family (the IPTs) and the gene family key to the destruction of cytokinins (the CKXs), as well as other gene families potentially affected by changing cytokinin levels. These included key meristem genes (WUS and BAM1) and the transporter gene families, sucrose transporters (SUTs) and amino acid permeases (AAPs). We used reverse transcription quantitative PCR (RT-qPCR) to monitor gene expression in the vegetative meristem and in pre- and post-fertilisation young pea fruits. PsWUS expression was specific to the shoot apical meristem while PsBAM1 was highly expressed in the shoot apical meristem (SAM) but was also expressed at a low level in the young fruit. Differential expression was shown between genes and within gene families for IPT, CKX, SUT, and AAP. PsCKX7 showed strong gene family member-specific expression in the SAM, and was also expressed in young pea fruits. We suggest that PsCKX7 is a potential target for downregulation via molecular breeding or gene editing. PMID:28212324

  16. Effect of Light Intensity on Efficiency of Carbon Dioxide and Nitrogen Reduction in Pisum sativum L. 1

    PubMed Central

    Bethlenfalvay, Gabor J.; Phillips, Donald A.

    1977-01-01

    Photosynthetic efficiency, primary productivity, and N2 reduction were determined in peas (Pisum sativum L. var. Alaska) grown at light intensities ranging from severely limiting to saturating. Plants grown under higher light intensities showed greater carboxylation and light capture potential and higher rates of net C exchange. Uptake of N2, computed from measured C2H2 reduction and H2 evolution rates, also increased with growth light intensity, while the previously proposed relative efficiency of N2 fixation, based on these same parameters, declined. The plot of N/C ratios (total nitrogen content/plant dry weight) increased hyperbolically with light intensity, and the plot of N2/CO2 uptake ratios (N2 uptake rate/net CO2 uptake rate) increased linearly. Both plots extrapolated to the light compensation point. The data indicate that the relative efficiency of N2 fixation is not necessarily correlated with maximum plant productivity and that evaluation of a plant's capacity to reduce N2 is related directly to concurrent CO2 reduction. A measure of whole plant N2 fixation efficiency based on the N2/CO2 uptake ratio is proposed. PMID:16660203

  17. Root and Nodule Growth in Pisum sativum L. in Relation to Photosynthesis: Analysis Using 13C‐labelling

    PubMed Central

    VOISIN, A. S.; SALON, C.; JEUDY, C.; WAREMBOURG, F. R.

    2003-01-01

    The effect of the nitrogen source (gaseous nitrogen, N2, or nitrate ions, NO3–) on the use of carbon (C) for root and nodule growth of pea (Pisum sativum L.) was investigated using 13C‐labelling of assimilated CO2 at various stages of growth. Nitrate supply and growing conditions (sowing dates, air CO2 concentration) were varied to alter photosynthetic rates. Nodules are the sink with the highest demand for C in both the vegetative and flowering stages, growing at the expense of shoot and root in the vegetative stage, but only at the expense of roots at flowering. Until flowering, the addition of C into root and nodule biomass was linearly related to pre‐existing biomass, thus determining net sink strengths which decreased with root and nodule age. Nodule growth patterns did not depend on the N source, whereas root growth was increased by nitrate when nodule biomass was low. At seed filling, the increase in C of biomass of the root system was no longer related to pre‐existing biomass and C was preferentially diverted to roots of plants assimilating nitrate, or to nodules for plants fixing N2. PMID:14507741

  18. Stomatal Response and Leaf Injury of Pisum sativum L. with SO2 and O3 Exposures 12

    PubMed Central

    Olszyk, David M.; Tibbitts, Theodore W.

    1981-01-01

    Plants of Pisum sativum L. `Alsweet' were grown under a controlled environment and exposed to SO2 and O3 to determine whether changes in stomatal aperture during exposure were related to subsequent leaf injury. Stomata consistently closed with injurious levels of SO2 and O3. Measurements with diffusion porometers demonstrated ≃75 and 25% lower conductance with SO2 and O3 exposures, respectively, compared to the conductance of control plants. Stomata also showed a closing response with noninjurious levels of SO2 but an opening response with noninjurious levels of O3. Stomata closed to the same degree with combinations of SO2 plus O3 as with SO2 alone. Stomata of expanding leaves closed more during pollutant exposures than stomata of expanded leaves. The abaxial and adaxial stomata both exhibited closure with SO2 and combinations of SO2 plus O3, but abaxial stomata tended to close and adaxial stomata tended to open with exposure to O3 alone. The changes in stomatal aperture were not closely correlated with the amount of leaf injury produced by different pollutant levels. Stomata closed, not only with exposure to pollutant levels that caused severe necrosis, but also with levels that caused only a trace of injury. There was no evidence of a reduced amount of closure or even stomatal opening with combinations of SO2 and O3 compared to plants exposed to the pollutants alone to explain the large amount of injury to plants exposed to pollutant combinations. PMID:16661710

  19. Gibberellic Acid-Promoted Lignification and Phenylalanine Ammonia-lyase Activity in a Dwarf Pea (Pisum sativum) 1

    PubMed Central

    Cheng, Christina K.-C.; Marsh, H. V.

    1968-01-01

    The effects of gibberellic acid on lignification in seedlings of a dwarf and a tall cultivar of pea (Pisum sativum) grown under red or white light or in the darkness, were studied. Gibberellic acid (10−6-10−4 m) promoted stem elongation in both light and dark and increased the percentage of lignin in the stems of the light-grown dwarf pea. The gibberellin had no effect on the lignin content of the tall pea although high concentrations (10−4 m) promoted growth of the tall plants. Time course studies indicated that the enhanced lignification in the gibberellin-treated dwarf plants occurred only after a lag period of several days. It was concluded that gibberellic acid-enhanced ligmification had no direct relation to gibberellic acid-promoted growth. The activity of phenylalanine ammonia-lyase (E.C. 4.3.1.5) was higher in gibberellin-treated dwarf plants grown under white or red light than in untreated dwarf plants. Gibberellic acid had no detectable effect on the activity of this enzyme when the plants were grown in darkness, just as it had no effect on lignification under dark conditions. The data suggest that in gibberellin-deficient peas the activity of phenylalanine ammonia-lyase is one of the limiting factors in lignification. PMID:16656968

  20. Molecular characterization of a distinct monopartite begomovirus associated with betasatellites and alphasatellites infecting Pisum sativum in Nepal.

    PubMed

    Shahid, M S; Pudashini, B J; Khatri-Chhetri, G B; Briddon, R W; Natsuaki, K T

    2017-04-01

    Pea (Pisum sativum) plants exhibiting leaf distortion, yellowing, stunted growth and reduction in leaf size from Rampur, Nepal were shown to be infected by a begomovirus in association with betasatellites and alphasatellites. The begomovirus associated with the disease showed only low levels of nucleotide sequence identity (<91%) to previously characterized begomoviruses. This finding indicates that the pea samples were infected with an as yet undescribed begomovirus for which the name Pea leaf distortion virus (PLDV) is proposed. Two species of betasatellite were identified in association with PLDV. One group of sequences had high (>78%) nucleotide sequence identity to isolates of Ludwigia leaf distortion betasatellite (LuLDB), and the second group had less than 78% to all other betasatellite sequences. This showed PLDV to be associated with either LuLDB or a previously undescribed betasatellite for which the name Pea leaf distortion betasatellite is proposed. Two types of alphasatellites were identified in the PLDV-infected pea plants. The first type showed high levels of sequence identity to Ageratum yellow vein alphasatellite, and the second type showed high levels of identity to isolates of Sida yellow vein China alphasatellite. These are the first begomovirus, betasatellites and alphasatellites isolated from pea.

  1. Remote sensing study of the influence of herbicides on the spectral reflectance of pea plant leaves (Pisum sativum L.)

    NASA Astrophysics Data System (ADS)

    Krezhova, D.; Alexieva, V.; Yanev, T.; Ivanov, S.

    Results from a remote sensing study of spectral reflectance of leaves of pea plants Pisum sativum L treated by the herbicides atrazine 2 4-D glyphosate fluridone and chlorsulfuron are reported According to the classification of the Herbicide Action Committee reflecting their mode of action they belong to different groups photosystem II bloker - C1 atrazine synthetic auxins - O 2 4-D inhibition of EPSP synthase - G glyphosate photobleaching - F1 fluridone and inhibition of acetoctate synthase - B chlorsulfuron The plants studied were grown hydroponically in a growth chamber in a nutritious medium to which every herbicide was added at three low concentrations 1 mu M 0 1 mu M and 0 01 mu M with respect to the field dose applied in the agricultural practice The spectral measurements of the leaf spectral reflectance were carried out in laboratory using a multichannel spectrometer in the visible and near infrared regions of the spectrum 480 div 810 nm Data was registered in 128 channels at a high spectral resolution of 2 6 nm halfwidth and a spatial resolution of 2 mm 2 The reflectance spectra were obtained from the leaf-reflected radiation referenced against a standard white screen To assess the changes arising in the leaf spectral reflectance under the herbicide action the developed by us approach based on discriminant analysis and other statistical methods was applied The spectral reflectance characteristics SRC were investigated in three spectral intervals 520 div 580 nm region of maximal

  2. Non-host disease resistance response in pea (Pisum sativum) pods: Biochemical function of DRR206 and phytoalexin pathway localization.

    PubMed

    Seneviratne, Herana Kamal; Dalisay, Doralyn S; Kim, Kye-Won; Moinuddin, Syed G A; Yang, Hong; Hartshorn, Christopher M; Davin, Laurence B; Lewis, Norman G

    2015-05-01

    Continually exposed to potential pathogens, vascular plants have evolved intricate defense mechanisms to recognize encroaching threats and defend themselves. They do so by inducing a set of defense responses that can help defeat and/or limit effects of invading pathogens, of which the non-host disease resistance response is the most common. In this regard, pea (Pisum sativum) pod tissue, when exposed to Fusarium solani f. sp. phaseoli spores, undergoes an inducible transcriptional activation of pathogenesis-related genes, and also produces (+)-pisatin, its major phytoalexin. One of the inducible pathogenesis-related genes is Disease Resistance Response-206 (DRR206), whose role in vivo was unknown. DRR206 is, however, related to the dirigent protein (DP) family. In this study, its biochemical function was investigated in planta, with the metabolite associated with its gene induction being pinoresinol monoglucoside. Interestingly, both pinoresinol monoglucoside and (+)-pisatin were co-localized in pea pod endocarp epidermal cells, as demonstrated using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging. In addition, endocarp epidermal cells are also the site for both chalcone synthase and DRR206 gene expression. Taken together, these data indicate that both (+)-pisatin and pinoresinol monoglucoside function in the overall phytoalexin responses. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. The flowering locus Hr colocalizes with a major QTL affecting winter frost tolerance in Pisum sativum L.

    PubMed

    Lejeune-Hénaut, I; Hanocq, E; Béthencourt, L; Fontaine, V; Delbreil, B; Morin, J; Petit, A; Devaux, R; Boilleau, M; Stempniak, J J; Thomas, M; Lainé, A L; Foucher, F; Baranger, A; Burstin, J; Rameau, C; Giauffret, C

    2008-05-01

    An understanding of the genetic determinism of frost tolerance is a prerequisite for the development of frost tolerant cultivars for cold northern areas. In legumes, it is not known to which extent vernalization requirement or photoperiod responsiveness are necessary for the development of frost tolerance. In pea (Pisum sativum L.) however, the flowering locus Hr is suspected to influence winter frost tolerance by delaying floral initiation until after the main winter freezing periods have passed. The objective of this study was to dissect the genetic determinism of frost tolerance in pea by QTL analysis and to assess the genetic linkage between winter frost tolerance and the Hr locus. A population of 164 recombinant inbred lines (RILs), derived from the cross Champagne x Terese was evaluated both in the greenhouse and in field conditions to characterize the photoperiod response from which the allele at the Hr locus was inferred. In addition, the population was also assessed for winter frost tolerance in 11 field conditions. Six QTL were detected, among which three were consistent among the different experimental conditions, confirming an oligogenic determinism of frost tolerance in pea. The Hr locus was found to be the peak marker for the highest explanatory QTL of this study. This result supports the hypothesis of the prominent part played by the photoperiod responsiveness in the determinism of frost tolerance for this species. The consistency of three QTL makes these positions interesting targets for marker-assisted selection.

  4. Efficient production of human acidic fibroblast growth factor in pea (Pisum sativum L.) plants by agroinfection of germinated seeds

    PubMed Central

    2011-01-01

    Background For efficient and large scale production of recombinant proteins in plants transient expression by agroinfection has a number of advantages over stable transformation. Simple manipulation, rapid analysis and high expression efficiency are possible. In pea, Pisum sativum, a Virus Induced Gene Silencing System using the pea early browning virus has been converted into an efficient agroinfection system by converting the two RNA genomes of the virus into binary expression vectors for Agrobacterium transformation. Results By vacuum infiltration (0.08 Mpa, 1 min) of germinating pea seeds with 2-3 cm roots with Agrobacteria carrying the binary vectors, expression of the gene for Green Fluorescent Protein as marker and the gene for the human acidic fibroblast growth factor (aFGF) was obtained in 80% of the infiltrated developing seedlings. Maximal production of the recombinant proteins was achieved 12-15 days after infiltration. Conclusions Compared to the leaf injection method vacuum infiltration of germinated seeds is highly efficient allowing large scale production of plants transiently expressing recombinant proteins. The production cycle of plants for harvesting the recombinant protein was shortened from 30 days for leaf injection to 15 days by applying vacuum infiltration. The synthesized aFGF was purified by heparin-affinity chromatography and its mitogenic activity on NIH 3T3 cells confirmed to be similar to a commercial product. PMID:21548923

  5. The effect of different alleles at the r locus on the synthesis of seed storage proteins in Pisum sativum.

    PubMed

    Turner, S R; Barratt, D H; Casey, R

    1990-05-01

    Rocket immunoelectrophoresis was used to measure the accumulation of storage proteins in developing cotyledons of two Pisum sativum (pea) genotypes, that were close to isogenic except for the nature of the allele at the r locus. There was a marked decrease in legumin accumulation in the rr (wrinkled-seeded) genotype compared to the RR (round-seeded) genotype. The accumulation of vicilin did not differ greatly between the two genotypes. Pulse-labelling studies indicated that the differences in rates of accumulation of legumin between the rr and RR genotypes were a consequence of differences in rates of protein synthesis. Measurements of relative amounts of specific mRNAs, using cDNA clones as probes, showed lower amounts of legumin mRNA in developing cotyledons of the rr, compared to the RR, genotype. Both vicilin mRNAs and convicilin mRNA, the latter of which shows a similar temporal pattern of expression to those of the major legumin species, are relatively unaffected by the nature of the allele at the r locus. Nuclear run-on transcription experiments indicated no differences in the rate of synthesis of legumin transcripts in the rr and RR near-isolines. The consequences of homozygosity for the r allele on storage protein mRNA levels in vitro may be mimicked by manipulating the sucrose concentration of the culture medium.

  6. Differential Gene Expression in the Meristem and during Early Fruit Growth of Pisum sativum L. Identifies Potential Targets for Breeding.

    PubMed

    Smitha Ninan, Annu; Shah, Anish; Song, Jiancheng; Jameson, Paula E

    2017-02-16

    For successful molecular breeding it is important to identify targets to the gene family level, and in the specific species of interest, in this case Pisum sativum L. The cytokinins have been identified as a key breeding target due to their influence on plant architecture, and on seed size and sink activity. We focused on the cytokinin biosynthetic gene family (the IPTs) and the gene family key to the destruction of cytokinins (the CKXs), as well as other gene families potentially affected by changing cytokinin levels. These included key meristem genes (WUS and BAM1) and the transporter gene families, sucrose transporters (SUTs) and amino acid permeases (AAPs). We used reverse transcription quantitative PCR (RT-qPCR) to monitor gene expression in the vegetative meristem and in pre- and post-fertilisation young pea fruits. PsWUS expression was specific to the shoot apical meristem while PsBAM1 was highly expressed in the shoot apical meristem (SAM) but was also expressed at a low level in the young fruit. Differential expression was shown between genes and within gene families for IPT, CKX, SUT, and AAP. PsCKX7 showed strong gene family member-specific expression in the SAM, and was also expressed in young pea fruits. We suggest that PsCKX7 is a potential target for downregulation via molecular breeding or gene editing.

  7. Purification, properties and amino acid sequence of a low-Mr abundant seed protein from pea (Pisum sativum L.).

    PubMed

    Gatehouse, J A; Gilroy, J; Hoque, M S; Croy, R R

    1985-01-01

    The seeds of pea (Pisum sativum L.) contain several proteins in the albumin solubility fraction that are significant components of total cotyledonary protein (5-10%) and are accumulated in developing seeds concurrently with storage-protein synthesis. One of these proteins, of low Mr and designated 'Psa LA', has been purified, characterized and sequenced. Psa LA has an Mr of 11000 and contains polypeptides of Mr 6000, suggesting that the protein molecules are dimeric. The amino acid sequence contains 54 residues, with a high content (10/54) of asparagine/aspartate. It has no inhibitory action towards trypsin or chymotrypsin, and is distinct from the inhibitors of those enzymes found in pea seeds, nor does it inhibit hog pancreatic alpha-amylase. The protein contains no methionine, but significant amounts of cysteine (four residues per polypeptide), suggesting a possible role as a sulphur storage protein. However, its sequence is not homologous with low-Mr (2S) storage proteins from castor bean (Ricinus communis) or rape (Brassica napus). Psa LA therefore represents a new type of low-Mr seed protein.

  8. Pea (Pisum sativum L.) seed albumin extracts show anti-inflammatory effect in the DSS model of mouse colitis.

    PubMed

    Utrilla, Ma Pilar; Peinado, Ma Jesus; Ruiz, Raquel; Rodriguez-Nogales, Alba; Algieri, Francesca; Rodriguez-Cabezas, Ma Elena; Clemente, Alfonso; Galvez, Julio; Rubio, Luis A

    2015-04-01

    This study investigates the preventive effects of two pea (Pisum sativum) seed albumin extracts, either in the presence (pea seed extract [PSE]) or absence (albumin fraction from PSE [AF-PSE]) of soluble polysaccharides, in the dextran sodium sulfate (DSS) induced colitis in mice. Male C57BL/6J mice were assigned to five groups: one noncolitic and four colitic. Colitis was induced by incorporating DSS (3.5%) in the drinking water for 4 days, after which DSS was removed. Treated groups received orally PSE (15 g/kg⋅day), or AF-PSE (1.5 g/kg⋅day), or pure soy Bowman-Birk inhibitor (BBI; 50 mg/kg⋅day), starting 2 wk before colitis induction, and maintained for 9 days after. All treated groups showed intestinal anti-inflammatory effect, evidenced by reduced microscopic histological damage in comparison with untreated colitic mice. The treatments ameliorated the colonic mRNA expression of different proinflammatory markers: cytokines, inducible enzymes, metalloproteinases, adhesion molecules, and toll-like receptors, as well as proteins involved in maintaining the epithelial barrier function. Furthermore, the administration of PSE, AF-PSE, or soy BBI restored bacterial counts, partially or totally, to values in healthy mice. PSE and AF-PSE ameliorated DSS-induced damage to mice, their effects being due, at least partially, to the presence of active BBI. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Foliar Application of Plant Growth-Promoting Rhizobacteria Increases Antifungal Compounds in Pea (Pisum sativum) Against Erysiphe pisi

    PubMed Central

    Bahadur, A.; Sarma, B. K.; Singh, D. P.; Singh, K. P.; Singh, A.

    2007-01-01

    Systemic effect of two plant growth-promoting rhizobacterial (PGPR) strains,viz., Pseudomonas fluorescens (Pf4) and P. aeruginosa (Pag), was evaluated on pea (Pisum sativum) against the powdery mildew pathogen Erysiphe pisi. Foliar spray of the two PGPR strains was done on specific nodal leaves of pea and conidial germination of E. pisi was observed on other nodal leaves,distal to the treated ones. Conidial germination was reduced on distant leaves and at the same time,specific as well as total phenolic compounds increased in the leaves distal to those applied with PGPR strains,thereby indicating a positive correlation. The strains induced accumulation of phenolic compounds in pea leaves and the amount increased when such leaves were get inoculated with E. pisi conidia. Between the two strains, Pag was found to be more effective than Pf4 as its effect was more persistent in pea leaves. Foliar application of PGPR strains for the control of powdery mildew of pea is demonstrated in vitro while correlating it with the increased accumulation of plant phenolics. PMID:24015083

  10. Expression of small heat shock protein (sHSP) genes in the garden pea (Pisum sativum) under slow horizontal clinorotation.

    PubMed

    Talalaiev, Oleksandr; Korduym, Elizabeth

    2014-04-30

    Plant cells respond to stress conditions, such as high temperatures, by synthesizing small heat shock proteins (sHSPs). sHSPs are molecular chaperones that assist in protein folding and prevent irreversible protein aggregation. Although many sHSP genes are temperature-inducible, other variables, such as altered gravity, can induce significant changes in plant cell gene expression. Furthermore, not all subfamilies of sHSP genes share the same expression pattern. The objective of our research was to determine the effect of simulated microgravity (clinorotation) on the expression of sHSP gene subfamilies with different subcellular locations in etiolated pea (Pisum sativum) seedlings. sHSP gene expression levels were examined using quantitative real-time RT-PCR (qPCR). qPCR results demonstrated that sHSP genes were constitutively expressed in seedlings. High temperatures increased the expression of sHSP genes by several thousand-fold. However, simulated microgravity did not have any significant effects on sHSP gene expression.

  11. Massive Analysis of cDNA Ends (MACE) for transcript-based marker design in pea (Pisum sativum L.).

    PubMed

    Zhernakov, Aleksandr; Rotter, Björn; Winter, Peter; Borisov, Alexey; Tikhonovich, Igor; Zhukov, Vladimir

    2017-03-01

    Aimed at gene-based markers design, we generated and analyzed transcriptome sequencing datasets for six pea (Pisum sativum L.) genetic lines that have not previously been massively genotyped. Five cDNA libraries obtained from nodules or nodulated roots of genetic lines Finale, Frisson, Sparkle, Sprint-2 and NGB1238 were sequenced using a versatile 3'-RNA-seq protocol called MACE (Massive Analysis of cDNA Ends). MACE delivers a single next-generation sequence from the 3'-end of each individual cDNA molecule that precisely quantifies the respective transcripts. Since the contig generated from the 3'-end of the cDNA by assembling all sequences encompasses the highly polymorphic 3'-untranslated region (3'-UTR), MACE efficiently detects single nucleotide variants (SNVs). Mapping MACE reads to the reference nodule transcriptome assembly of the pea line SGE (Transcriptome Shotgun Assembly GDTM00000000.1) resulted in characterization of over 34,000 polymorphic sites in more than 9700 contigs. Several of these SNVs were located within recognition sequences of restriction endonucleases which allowed the design of co-dominant CAPS markers for the particular transcript. Cleaned reads of sequenced libraries are available from European Nucleotide Archive (http://www.ebi.ac.uk/) under accessions PRJEB18101, PRJEB18102, PRJEB18103, PRJEB18104, PRJEB17691.

  12. Stress induced β subunit of heterotrimeric G-proteins from Pisum sativum interacts with mitogen activated protein kinase

    PubMed Central

    Bhardwaj, Deepak; Sheikh, Arsheed Hussain; Sinha, Alok Krishna

    2011-01-01

    We here report in Pisum sativum system a novel protein-protein interaction of β-subunit of heterotrimeric G-proteins (PsGβ) with a Mitogen activated protein kinase (PsMPK3) during cDNA library screening by yeast-two-hybrid assay. The transcript of these two genes also showed co-regulation under abscisic acid (ABA) and methyl jasmonate (MeJA) treatments. The protein-protein interaction was further validated by performing one-to-one interaction and β-galactosidase assay in yeast system. β-subunit of G-proteins from a heterologous system Oryzae sativa also showed interaction with PsMPK3. The interaction between PsGβ and PsMPK3 was further confirmed by in vitro protein-protein interaction. This suggested that MPK3 function as effector molecule for Gβ, which may helps in the regulation of stomatal functioning. These findings also provide an evidence for a possible cross-talk between MPK3 and G-protein-mediated signaling pathways in plants. PMID:21350337

  13. Efficient production of human acidic fibroblast growth factor in pea (Pisum sativum L.) plants by agroinfection of germinated seeds.

    PubMed

    Fan, Yajun; Li, Wei; Wang, Junjie; Liu, Jingying; Yang, Meiying; Xu, Duo; Zhu, Xiaojuan; Wang, Xingzhi

    2011-05-06

    For efficient and large scale production of recombinant proteins in plants transient expression by agroinfection has a number of advantages over stable transformation. Simple manipulation, rapid analysis and high expression efficiency are possible. In pea, Pisum sativum, a Virus Induced Gene Silencing System using the pea early browning virus has been converted into an efficient agroinfection system by converting the two RNA genomes of the virus into binary expression vectors for Agrobacterium transformation. By vacuum infiltration (0.08 Mpa, 1 min) of germinating pea seeds with 2-3 cm roots with Agrobacteria carrying the binary vectors, expression of the gene for Green Fluorescent Protein as marker and the gene for the human acidic fibroblast growth factor (aFGF) was obtained in 80% of the infiltrated developing seedlings. Maximal production of the recombinant proteins was achieved 12-15 days after infiltration. Compared to the leaf injection method vacuum infiltration of germinated seeds is highly efficient allowing large scale production of plants transiently expressing recombinant proteins. The production cycle of plants for harvesting the recombinant protein was shortened from 30 days for leaf injection to 15 days by applying vacuum infiltration. The synthesized aFGF was purified by heparin-affinity chromatography and its mitogenic activity on NIH 3T3 cells confirmed to be similar to a commercial product.

  14. Effect of antifungal genes expressed in transgenic pea (Pisum sativum L.) on root colonization with Glomus intraradices.

    PubMed

    Hassan, Fathi; Noorian, Mojgan Sharifi; Jacobsen, Hans-Jörg

    2012-01-01

    Pathogenic fungi have always been a major problem in agriculture. One of the effective methods for controlling pathogen fungi to date is the introduction of resistance genes into the genome of crops. It is interesting to find out whether the induced resistance in crops will have a negative effect on non-target organisms such as root colonization with the AM fungi.   The objective of the present research was to study the influence of producing antifungal molecules by four transgenic pea (Pisum sativum L.) lines expressing PGIP gene from raspberry, VST-stilbene synthase from vine, a hybrid of PGIP/VST and bacterial Chitinase gene (Chit30) from Streptomyces olivaceoviridis respectively on the colonization potential of Glomus intraradices. Four different experiments were done in greenhouse and climate chamber, colonization was observed in all replications. The following parameters were used for evaluation: frequency of mycorrhization, the intensity of mycorrhization, the average presence of arbuscules within the colonized areas and the presence of arbuscules in the whole root system which showed insignificant difference between transgenic and non-transgenic plants. The root/shoot ratio exhibited different values according to the experiment condition. Compared with negative non-transgenic control all transgenic lines showed the ability to establish symbiosis and the different growth parameters had insignificant effect due to mycorrhization. The results of the present study proved that the introduced pathogen resistance genes did not affect the mycorrhization allocations in pea.

  15. Fatty acid alpha-dioxygenase from Pisum sativum: temporal and spatial regulation during germination and plant development.

    PubMed

    Meisner, Anke K; Saffert, Alexander; Schreier, Peter; Schön, Astrid

    2009-03-01

    alpha-Dioxygenases are expressed in plants in response to biotic and abiotic stress. They catalyze the enantioselective 2-hydroperoxidation of long-chain fatty acids, the initial step of the alpha-oxidation pathway of fatty acids in plants. In this study, the complete cDNA of an alpha-dioxygenase from germinating pea seeds (Pisum sativum) is presented. The deduced amino acid sequence establishes that the enzyme belongs to the recently characterized family of alpha-dioxygenating enzymes in plants. We also present the first systematic study on the expression of alpha-dioxygenase in germinating and developing pea plants. During germination, alpha-dioxygenase mRNA accumulates in the cotyledons and the embryonic axis of pea seeds de novo. In developing pea plants, the transcript is detected almost exclusively in roots. The accumulation of alpha-dioxygenase protein parallels transcript accumulation in that it is abundant in germinating as well as young plant tissue, and correlates with loss of mRNA during plant maturation. alpha-Dioxygenase enzymatic activity in plant extracts is highest in cotyledons during imbibition. In the embryonic axis and roots of developing plants comparable activity levels are observed, whereas in shoots little alpha-oxidation activity is detected. With this contribution, we present information on the temporal and spatial expression of alpha-dioxygenase during plant germination and development, supporting the hypothesis that the alpha-oxidation pathway of fatty acids plays a role during plant developmental processes.

  16. The role of the epidermis in auxin-induced and fusicoccin-induced elongation of Pisum sativum stem segments.

    PubMed

    Brummell, D A; Hall, J L

    1980-12-01

    The effects of peeling and wounding on the indole-3-acetic acid (IAA) and fusicoccin (FC) growth response of etiolated Pisum sativum L. cv. Alaska stem tissue were examined. Over a 5 h growth period, peeling was found to virtually eliminate the IAA response, but about 30% of the FC response remained. In contrast, unpeeled segments wounded with six vertical slits exhibited significant responses to both IAA and FC, indicating that peeling does not act by damaging the tissue. Microscopy showed that the epidermis was removed intact and that the underlying tissue was essentially undamaged. Neither the addition of 2% sucrose to the incubation medium nor the use of a range of IAA concentrations down to 10(-8) M restored IAA-induced growth in peeled segments, suggesting that lack of osmotic solutes and supra-optimal uptake of IAA were not important factors over this time period. It is concluded that, although the possibility remains that peeling merely allows leakage of hydrogen ions into the medium, it seems more likely that peeling off the epidermis removes the auxin responsive tissue.

  17. Distribution and Properties of a Potassium-dependent Asparaginase Isolated from Developing Seeds of Pisum sativum and Other Plants 1

    PubMed Central

    Sodek, Ladaslav; Lea, Peter J.; Miflin, Benjamin J.

    1980-01-01

    Asparaginase (EC 3.5.1.1) was isolated from the developing seed of Pisum sativum. The enzyme is dependent upon the presence of K+ for activity, although Na+ and Rb+ may substitute to a lesser extent. Maximum activity was obtained at K+ concentrations above 20 millimolar. Potassium ions protected the enzyme against heat denaturation. The enzyme has a molecular weight of 68,300. Asparaginase activity developed initially in the testa, with maximum activity (3.6 micromoles per hour per seed) being present 13 days after flowering. Maximum activity (1.2 micromoles per hour per seed) did not develop in the cotyledon until 21 days after flowering. Glutamine synthetase and glutamate dehydrogenase were also present in the testae and cotyledons but maximum activity developed later than that of asparaginase. Potassium-dependent asparaginase activity was also detected in the developing seeds of Vicia faba, Phaseolus multiflorus, Zea mays, Hordeum vulgare, and two Lupinus varieties. No stimulation of activity was detected with the enzyme isolated from Lupinus polyphyllus, which has previously been shown to contain a K+-independent enzyme. PMID:16661136

  18. Apparent equilibrium constant and mass-action ratio for sucrose-phosphate synthase in seeds of Pisum sativum.

    PubMed Central

    Lunn, J E; ap Rees, T

    1990-01-01

    The aim of this work was to use preparations from germinating seeds of Pisum sativum to determine the apparent equilibrium constant of the reaction catalysed by sucrose-phosphate synthase (EC 2.4.1.14) and to compare this with the mass-action ratio of the reaction in the seeds. The apparent equilibrium constant ranged from 5.3 at 0.25 mM-MgCl2, pH 7.0, to 62 at 10 mM-MgCl2, pH 7.5. The sucrose phosphate content of the seeds, 23 nmol/g fresh wt., was determined by separating sucrose phosphate from sucrose by ion-exchange chromatography and then measuring the sucrose released by alkaline phosphatase. Comparison of equilibrium constants and mass-action ratios in the cotyledons of 38 h-germinated seeds showed that the reactions catalysed by glucose-6-phosphate isomerase, phosphoglucomutase and UDP-glucose pyrophosphorylase are close to equilibrium, and those catalysed by sucrose-phosphate synthase and sucrose phosphatase are considerably displaced from equilibrium in vivo. PMID:2140258

  19. Pinolide, a new nonenolide produced by Didymella pinodes , the causal agent of ascochyta blight on Pisum sativum.

    PubMed

    Cimmino, Alessio; Andolfi, Anna; Fondevilla, Sara; Abouzeid, Mohamed A; Rubiales, Diego; Evidente, Antonio

    2012-05-30

    An aggressive isolate of Didymella pinodes isolated from pea ( Pisum sativum ) produced four different metabolites in vitro. The metabolites isolated from the culture filtrates were characterized by spectroscopic and optical methods. A new nonenolide, named pinolide, was isolated and characterized as (2S*,7R*,8S*,5E,9R*)-2,7,8-trihydroxy-9-propyl-5-nonen-9-olide. Pinolidoxin, the main toxin produced by D. pinodes, was also isolated together with two other closely related nonenolides, identified as herbarumin II and 2-epi-herbarumin II. Herbarumin II and 2-epi-herbarumin II have been previously isolated from the fungi Phoma herbarum and Paraphaeosphaeria recurvifoliae , respectively, but described here to be isolated for the first time from D. pinodes. When tested on leaves of the host plant and other legumes and weeds, pinolidoxin was phytotoxic in all of the plant species, whereas the other three nonenolides did not produce any symptoms. The importance of the stereochemistry of the hydroxy group at C-7 on phytotoxicity also is discussed.

  20. The movement of 2,4-dichlorophenoxy acetic acid in root segments of Pisum sativum L.

    PubMed

    Wilkins, H; Wilkins, M B

    1975-01-01

    The movement of 2,4-Dichlorophenoxy acetic acid (2,4-D) through subapical segments of the primary roots of Pisum seedlings has been investigated using [1-(14)C]2,4-D.Donation of [1-(14)C]2,4-D to the apical or basal ends of Pisum root segments at 25°C in darkness revealed a preferential movement of the compound towards the root apex i.e. an acropetal polarisation. Thus the movement of [1-(14)C]2,4-D into receiver blocks applied to the apical ends of the segments is greater than that into receiver blocks applied to the basal ends of the segments The low level of basipetal transport appears to be associated with a restriction of the movement of [1-(14)C]2,4-D to the half of the segment nearest the donor block.Acropetal transport of 2,4-D is faster than basipetal transport in root segments maintained at 15° and 35° C but is slower than basipetal transport if the segments are maintained at 25°C. Maximum velocitees are 0.71 and 0.83 mm h(-1) for acropetal and basipetal transport respectively.Evidence from experiments carried out (a) in an anaerobic environment in the presence or absence of sodium fluoride and (b) over a range of temperatures from 1-35°C, indicates that the movement of [1-(14)C]2,4-D is dependent on the metabolic activity of the Pisum root segments.Release of (14)CO2 during transport of [1-(14)C]2,4-D is small and supports chromatographic evidence that negligible degradation of the 2,4-D molecules takes place during transport through the root segments.

  1. Optimization of Agroinfiltration in Pisum sativum Provides a New Tool for Studying the Salivary Protein Functions in the Pea Aphid Complex.

    PubMed

    Guy, Endrick; Boulain, Hélène; Aigu, Yoann; Le Pennec, Charlotte; Chawki, Khaoula; Morlière, Stéphanie; Schädel, Kristina; Kunert, Grit; Simon, Jean-Christophe; Sugio, Akiko

    2016-01-01

    Aphids are piercing-sucking insect pests and feed on phloem sap. During feeding, aphids inject a battery of salivary proteins into host plant. Some of these proteins function like effectors of microbial pathogens and influence the outcome of plant-aphid interactions. The pea aphid (Acyrthosiphon pisum) is the model aphid and encompasses multiple biotypes each specialized to one or a few legume species, providing an opportunity to investigate the underlying mechanisms of the compatibility between plants and aphid biotypes. We aim to identify the aphid factors that determine the compatibility with host plants, hence involved in the host plant specialization process, and hypothesize that salivary proteins are one of those factors. Agrobacterium-mediated transient gene expression is a powerful tool to perform functional analyses of effector (salivary) proteins in plants. However, the tool was not established for the legume species that A. pisum feeds on. Thus, we decided to optimize the method for legume plants to facilitate the functional analyses of A. pisum salivary proteins. We screened a range of cultivars of pea (Pisum sativum) and alfalfa (Medicago sativa). None of the M. sativa cultivars was suitable for agroinfiltration under the tested conditions; however, we established a protocol for efficient transient gene expression in two cultivars of P. sativum, ZP1109 and ZP1130, using A. tumefaciens AGL-1 strain and the pEAQ-HT-DEST1 vector. We confirmed that the genes are expressed from 3 to 10 days post-infiltration and that aphid lines of the pea adapted biotype fed and reproduced on these two cultivars while lines of alfalfa and clover biotypes did not. Thus, the pea biotype recognizes these two cultivars as typical pea plants. By using a combination of ZP1109 and an A. pisum line, we defined an agroinfiltration procedure to examine the effect of in planta expression of selected salivary proteins on A. pisum fitness and demonstrated that transient expression of

  2. Optimization of Agroinfiltration in Pisum sativum Provides a New Tool for Studying the Salivary Protein Functions in the Pea Aphid Complex

    PubMed Central

    Guy, Endrick; Boulain, Hélène; Aigu, Yoann; Le Pennec, Charlotte; Chawki, Khaoula; Morlière, Stéphanie; Schädel, Kristina; Kunert, Grit; Simon, Jean-Christophe; Sugio, Akiko

    2016-01-01

    Aphids are piercing-sucking insect pests and feed on phloem sap. During feeding, aphids inject a battery of salivary proteins into host plant. Some of these proteins function like effectors of microbial pathogens and influence the outcome of plant–aphid interactions. The pea aphid (Acyrthosiphon pisum) is the model aphid and encompasses multiple biotypes each specialized to one or a few legume species, providing an opportunity to investigate the underlying mechanisms of the compatibility between plants and aphid biotypes. We aim to identify the aphid factors that determine the compatibility with host plants, hence involved in the host plant specialization process, and hypothesize that salivary proteins are one of those factors. Agrobacterium-mediated transient gene expression is a powerful tool to perform functional analyses of effector (salivary) proteins in plants. However, the tool was not established for the legume species that A. pisum feeds on. Thus, we decided to optimize the method for legume plants to facilitate the functional analyses of A. pisum salivary proteins. We screened a range of cultivars of pea (Pisum sativum) and alfalfa (Medicago sativa). None of the M. sativa cultivars was suitable for agroinfiltration under the tested conditions; however, we established a protocol for efficient transient gene expression in two cultivars of P. sativum, ZP1109 and ZP1130, using A. tumefaciens AGL-1 strain and the pEAQ-HT-DEST1 vector. We confirmed that the genes are expressed from 3 to 10 days post-infiltration and that aphid lines of the pea adapted biotype fed and reproduced on these two cultivars while lines of alfalfa and clover biotypes did not. Thus, the pea biotype recognizes these two cultivars as typical pea plants. By using a combination of ZP1109 and an A. pisum line, we defined an agroinfiltration procedure to examine the effect of in planta expression of selected salivary proteins on A. pisum fitness and demonstrated that transient expression of

  3. Nickel availability to pea (Pisum sativum L.) plants limits hydrogenase activity of Rhizobium leguminosarum bv. viciae bacteroids by affecting the processing of the hydrogenase structural subunits.

    PubMed

    Brito, B; Palacios, J M; Hidalgo, E; Imperial, J; Ruiz-Argüeso, T

    1994-09-01

    Rhizobium leguminosarum bv. viciae UPM791 induces the synthesis of an [NiFe] hydrogenase in pea (Pisum sativum L.) bacteroids which oxidizes the H2 generated by the nitrogenase complex inside the root nodules. The synthesis of this hydrogenase requires the genes for the small and large hydrogenase subunits (hupS and hupL, respectively) and 15 accessory genes clustered in a complex locus in the symbiotic plasmid. We show here that the bacteroid hydrogenase activity is limited by the availability of nickel to pea plants. Addition of Ni2+ to plant nutrient solutions (up to 10 mg/liter) resulted in sharp increases (up to 15-fold) in hydrogenase activity. This effect was not detected when other divalent cations (Zn2+, Co2+, Fe2+, and Mn2+) were added at the same concentrations. Determinations of the steady-state levels of hupSL-specific mRNA indicated that this increase in hydrogenase activity was not due to stimulation of transcription of structural genes. Immunoblot analysis with antibodies raised against the large and small subunits of the hydrogenase enzyme demonstrated that in the low-nickel situation, both subunits are mainly present in slow-migrating, unprocessed forms. Supplementation of the plant nutrient solution with increasing nickel concentrations caused the conversion of the slow-migrating forms of both subunits into fast-moving, mature forms. This nickel-dependent maturation process of the hydrogenase subunits is mediated by accessory gene products, since bacteroids from H2 uptake-deficient mutants carrying Tn5 insertions in hupG and hupK and in hypB and hypE accumulated the immature forms of both hydrogenase subunits even in the presence of high nickel levels.

  4. Nickel availability to pea (Pisum sativum L.) plants limits hydrogenase activity of Rhizobium leguminosarum bv. viciae bacteroids by affecting the processing of the hydrogenase structural subunits.

    PubMed Central

    Brito, B; Palacios, J M; Hidalgo, E; Imperial, J; Ruiz-Argüeso, T

    1994-01-01

    Rhizobium leguminosarum bv. viciae UPM791 induces the synthesis of an [NiFe] hydrogenase in pea (Pisum sativum L.) bacteroids which oxidizes the H2 generated by the nitrogenase complex inside the root nodules. The synthesis of this hydrogenase requires the genes for the small and large hydrogenase subunits (hupS and hupL, respectively) and 15 accessory genes clustered in a complex locus in the symbiotic plasmid. We show here that the bacteroid hydrogenase activity is limited by the availability of nickel to pea plants. Addition of Ni2+ to plant nutrient solutions (up to 10 mg/liter) resulted in sharp increases (up to 15-fold) in hydrogenase activity. This effect was not detected when other divalent cations (Zn2+, Co2+, Fe2+, and Mn2+) were added at the same concentrations. Determinations of the steady-state levels of hupSL-specific mRNA indicated that this increase in hydrogenase activity was not due to stimulation of transcription of structural genes. Immunoblot analysis with antibodies raised against the large and small subunits of the hydrogenase enzyme demonstrated that in the low-nickel situation, both subunits are mainly present in slow-migrating, unprocessed forms. Supplementation of the plant nutrient solution with increasing nickel concentrations caused the conversion of the slow-migrating forms of both subunits into fast-moving, mature forms. This nickel-dependent maturation process of the hydrogenase subunits is mediated by accessory gene products, since bacteroids from H2 uptake-deficient mutants carrying Tn5 insertions in hupG and hupK and in hypB and hypE accumulated the immature forms of both hydrogenase subunits even in the presence of high nickel levels. Images PMID:8071205

  5. Effects of Arsenite, Sulfite, and Sulfate on Photosynthetic Carbon Metabolism in Isolated Pea (Pisum sativum L., cv Little Marvel) Chloroplasts 1

    PubMed Central

    Marques, Ivano A.; Anderson, Louise E.

    1986-01-01

    Photosynthetic CO2-fixation in isolated pea (Pisum sativum L., cv Little Marvel) chloroplasts during induction is markedly inhibited by 0.4 millimolar sulfite. Sulfate at the same concentration has almost no effect. The 14CO2-fixation pattern indicates that the primary effect of sulfite is inhibition of the reaction catalyzed by ribulose bisphosphate carboxylase and a stimulation of export of intermediates out of the chloroplasts. Inhibition of light modulation of stromal enzyme activity does not appear to account for the toxicity of SO2 in this Pisum variety. Arsenite at 0.2 millimolar concentrations inhibits light activation and inhibits photosynthetic CO2 fixation. The 14CO2-fixation pattern indicates that the primary effect of arsenite is inhibition of light activation of reductive pentose phosphate pathway enzyme activity. PMID:16665056

  6. [Influence of calcium and rhizobial infections (Rhizobium leguminosarum) on the dynamics of nitric oxide (NO) content in roots of etiolated pea (Pisum sativum L.) seedlings].

    PubMed

    Glian'ko, A K; Ishchenko, A A; Stepanov, A V

    2014-01-01

    The effect of exogenous calcium (Ca2+) and rhizobial infections (Rhizobium leguminosarum bv viceae) on the dynamics of the level of nitric oxide (NO) was studied in cross cuts of roots of two-day-old etiolated pea seedlings (Pisum sativum L.) using a DAF-2DA fluorescent probe. Fluctuations of the NO level, indicating the presence of a rhythm in the generation of NO in roots, were observed during the incubation of seedlings in water, a CaCl2 solution, and with rhizobial infections. Exogenous factors (Ca2+ and two rhizobial stamms) change the time dynamics of the NO level in comparison with the control (water).

  7. The auxin conjugate indole-3-acetyl-aspartate affects responses to cadmium and salt stress in Pisum sativum L.

    PubMed

    Ostrowski, Maciej; Ciarkowska, Anna; Jakubowska, Anna

    2016-02-01

    The synthesis of IAA-amino acid conjugates is one of the crucial regulatory mechanisms for the control of auxin activity during physiological and pathophysiological responses. Indole-3-acetyl-aspartate (IAA-Asp) is a low molecular weight amide conjugate that predominates in pea (Pisum sativum L.) tissues. IAA-Asp acts as an intermediate during the auxin degradation pathway. However, some recent investigations suggest a direct signaling function of this conjugate in various processes. In this study, we examine the effect of 100 μM IAA-Asp alone and in combination with salt stress (160 mM NaCl) or heavy metal stress (250 μM CdCl2) on H2O2 concentration, protein carbonylation as well as catalase and ascorbate (APX) and guaiacol peroxidase (GPX) activities in 7-day-old pea seedlings. As revealed by spectrophotometric analyses, IAA-Asp increased the carbonylated protein level and reduced the H2O2 concentration. Moreover, IAA-aspartate potentiated the effect of both Cd(2+) ions and NaCl on the H2O2 level. The enzymatic activities (catalase and peroxidases) were examined using spectrophotometric and native-PAGE assays. IAA-Asp alone did not affect catalase activity, whereas the two peroxidases were regulated differently. IAA-Asp reduced the APX activity during 48h cultivation. APX activity was potentiated by IAA-Asp+NaCl after 48h. Guaiacol peroxidase activity was diminished by all tested compounds. Based on these results, we suggest that IAA-Asp can directly and specifically affect the pea responses to abiotic stress. Copyright © 2015 Elsevier GmbH. All rights reserved.

  8. Symbiotic N2 fixation activity in relation to C economy of Pisum sativum L. as a function of plant phenology.

    PubMed

    Voisin, A S; Salon, C; Jeudy, C; Warembourg, F R

    2003-12-01

    The relationships between symbiotic nitrogen fixation (SNF) activity and C fluxes were investigated in pea plants (Pisum sativum L. cv. Baccara) using simultaneous 13C and 15N labelling. Analysis of the dynamics of labelled CO2 efflux from the nodulated roots allowed the different components associated with SNF activity to be calculated, together with root and nodule synthetic and maintenance processes. The carbon costs for the synthesis of roots and nodules were similar and decreased with time. Carbon lost by turnover, associated with maintenance processes, decreased with time for nodules while it increased in the roots. Nodule turnover remained higher than root turnover until flowering. The effect of the N source on SNF was investigated using plants supplied with nitrate or plants only fixing N2. SNF per unit nodule biomass (nodule specific activity) was linearly related to the amount of carbon allocated to the nodulated roots regardless of the N source, with regression slopes decreasing across the growth cycle. These regression slopes permitted potential values of SNF specific activity to be defined. SNF activity decreased as the plants aged, presumably because of the combined effects of both increasing C costs of SNF (from 4.0 to 6.7 g C g-1 N) and the limitation of C supply to the nodules. SNF activity competed for C against synthesis and maintenance processes within the nodulated roots. Synthesis was the main limiting factor of SNF, but its importance decreased as the plant aged. At seed-filling, SNF was probably more limited by nodule age than by C supply to the nodulated roots.

  9. Effects of Glycolate Pathway Intermediates on Glycine Decarboxylation and Serine Synthesis in Pea (Pisum sativum L.) 1

    PubMed Central

    Shingles, Richard; Woodrow, Lorna; Grodzinski, Bernard

    1984-01-01

    Glycine decarboxylation and serine synthesis were studied in pea (Pisum sativum L.) leaf discs, in metabolically active intact chloroplasts, and in mitochondria isolated both partially by differential centrifugation (i.e. `crude') and by further purification on a Percoll gradient. Glycolate, glyoxylate, and formate reduced glycine decarboxylase activity (14CO2 and NH3 release) in the crude green-colored mitochondrial fractions, and in the leaf discs without markedly altering serine synthesis from [1-14C]glycine. Glycolate acted because it was converted to glyoxylate which behaves as a noncompetitive inhibitor (Ki = 5.1 ± 0.5 millimolar) on the mitochondrial glycine decarboxylation reaction in both crude and Percoll-purified mitochondria. In contrast, formate facilitates glycine to serine conversion by a route which does not involve glycine breakdown in the crude mitochondrial fraction and leaf discs. Formate does not alter the conversion of two molecules of glycine to one CO2, one NH3, and one serine molecule in the Percoll-purified mitochondria. In chloroplasts which were unable to break glycine down to CO2 and NH3, serine was labeled equally from [14C]formate and [1-14C]glycine. The maximum rate of serine synthesis observed in chloroplasts is similar to that in isolated metabolically active mitochondria. Formate does not appear to be able to substitute for the one-carbon unit produced during mitochondrial glycine breakdown but can facilitate serine synthesis from glycine in a chloroplast reaction which is probably a secondary one in vivo. PMID:16663485

  10. Enzymes of serine and glycine metabolism in leaves and non-photosynthetic tissues of Pisum sativum L.

    PubMed

    Walton, N J; Woolhouse, H W

    1986-01-01

    A comparative study is presented of the activities of enzymes of glycine and serine metabolism in leaves, germinated cotyledons and root apices of pea (Pisum sativum L.). Data are given for aminotransferase activities with glyoxylate, hydroxypyruvate and pyruvate, for enzymes associated with serine synthesis from 3-phosphoglycerate and for glycine decarboxylase and serine hydroxymethyltransferase. Aminotransferase activities differ between the tissues in that, firstly, appreciable transamination of serine, hydroxypyruvate and asparagine occurs only in leaf extracts and, secondly, glyoxylate is transaminated more actively than pyruvate in leaf extracts, whereas the converse is true of extracts of cotyledons and root apices. Alanine is the most active amino-group donor to both glyoxylate and hydroxypyruvate. 3-Phosphoglycerate dehydrogenase and glutamate: O-phosphohydroxypyruvate aminotransferase have comparable activities in all three tissues, except germinated cotyledons, in which the aminotransferase appears to be undetectable. Glycollate oxidase is virtually undetectable in the non-photosynthetic tissues and in these tissues the activity of glycerate dehydrogenase is much lower than that of 3-phosphoglycerate dehydrogenase. Glycine decarboxylase activity in leaves, measured in the presence of oxaloacetate, is equal to about 30-40% of the measured rate of CO2 fixation and is therefore adequate to account for the expected rate of photorespiration. The activity of glycine decarboxylase in the non-photosynthetic tissues is calculated to be about 2-5% of the activity in leaves and has the characteristics of a pyridoxal-and tetrahydrofolate-dependent mitochondrial reaction; it is stimulated by oxaloacetate, although not by ADP. In leaves, the measured activity of serine hydroxymethyltransferase is somewhat lower than that of glycine decarboxylase, whereas in root apices it is substantially higher. Differential centrifugation of extracts of root apices suggests that an

  11. Enzymology of Glutamine Metabolism Related to Senescence and Seed Development in the Pea (Pisum sativum L.) 1

    PubMed Central

    Storey, Richard; Beevers, Leonard

    1978-01-01

    The metabolism of glutamine in the leaf and subtended fruit of the aging pea (Pisum sativum L. cv. Burpeeana) has been studied in relation to changes in the protein, chlorophyll, and free amino acid content of each organ during ontogenesis. Glutamine synthetase [EC 6.3.1.2] activity was measured during development and senescence in each organ. Glutamate synthetase [EC 2.6.1.53] activity was followed in the pod and cotyledon during development and maturation. Maximal glutamine synthetase activity and free amino acid accumulation occurred together in the young leaf. Glutamine synthetase (in vitro) in leaf extracts greatly exceeded the requirement (in vivo) for reduced N in the organ. Glutamine synthetase activity, although declining in the senescing leaf, was sufficient (in vitro) to produce glutamine from all of the N released during protein hydrolysis (in vivo). Maximal glutamine synthetase activity in the pod was recorded 6 days after the peak accumulation of the free amino acids in this organ. In the young pod, free amino acids accumulated as glutamate synthetase activity increased. Maximal pod glutamate synthetase activity occurred simultaneously with maximal leaf glutamine synthetase activity, but 6 days prior to the corresponding maximum of glutamine synthetase in the pod. Cotyledonary glutamate synthetase activity increased during the assimilatory phase of embryo growth which coincided with the loss of protein and free amino acids from the leaf and pod; maximal activity was recorded simultaneously with maximal pod glutamine synthetase. We suggest that the activity of glutamine synthetase in the supply organs (leaf, pod) furnishes the translocated amide necessary for the N nutrition of the cotyledon. The subsequent activity of glutamate synthetase could provide a mechanism for the transfer of imported amide N to alpha amino N subsequently used in protein synthesis. In vitro measurements of enzyme activity indicate there was sufficient catalytic potential in

  12. Expression of the le Mutation in Young Ovaries of Pisum sativum and Its Effect on Fruit Development.

    PubMed Central

    Santes, C. M.; Hedden, P.; Sponsel, V. M.; Reid, J. B.; Garcia-Martinez, J. L.

    1993-01-01

    The effect of the le mutation on the growth and gibberellin (GA) content of developing fruits was investigated using the near-isogenic lines of Pisum sativum L. 205+ (LeLe) and 205- (lele). Although stem elongation is known to be reduced in 205- plants by approximately 65%, the growth of pods and seeds was unaffected by the le mutation. GA1, GA3, and GA20 stimulated parthenocarpic development of unpollinated ovaries on both 205+ and 205- plants. GA20 was less active on 205- ovaries than on 205+, whereas GA1 had similar, high activity in both lines. The activity of GA3 was even higher than that of GA1 in both lines. Decapitation of 205+ plants induced parthenocarpic development of unpollinated ovaries, but this treatment was much less effective on 205- plants. The contents of GA1 and GA8 in entire ovaries 6 d after anthesis, as well as in the pod and fertilized ovules, were substantially lower in 205- than in 205+ plants, whereas the reverse was true for the levels of GA20 and GA29. These results suggest that 3[beta]-hydroxylation of GA20 to GA1 is reduced in ovaries as well as in vegetative tissues. Thus, the le mutation appears to be expressed in young reproductive organs of the 205- line, even though it does not affect the fruit phenotype. Because the content of GA3 in the ovary was similar in the two lines, one explanation for the normal fruit size in the 205- line is that GA3 is the native regulator of pod growth. Alternatively, sufficient GA1 may still be produced in 205- fruits to maintain normal pod growth. PMID:12231727

  13. Expression of protein complexes and individual proteins upon transition of etioplasts to chloroplasts in pea (Pisum sativum).

    PubMed

    Kanervo, Eira; Singh, Munna; Suorsa, Marjaana; Paakkarinen, Virpi; Aro, Eveliina; Battchikova, Natalia; Aro, Eva-Mari

    2008-03-01

    The protein complexes of pea (Pisum sativum L.) etioplasts, etio-chloroplasts and chloroplasts were examined using 2D Blue Native/SDS-PAGE. The most prominent protein complexes in etioplasts were the ATPase and the Clp and FtsH protease complexes which probably have a crucial role in the biogenesis of etioplasts and chloroplasts. Also the cytochrome b(6)f (Cyt b(6)f) complex was assembled in the etioplast membrane, as well as Rubisco, at least partially, in the stroma. These complexes are composed of proteins encoded by both the plastid and nuclear genomes, indicating that a functional cross-talk exists between pea etioplasts and the nucleus. In contrast, the proteins and protein complexes that bind chlorophyll, with the PetD subunit and the entire Cyt b(6)f complex as an exception, did not accumulate in etioplasts. Nevertheless, some PSII core components such as PsbE and the luminal oxygen-evolvong complex (OEC) proteins PsbO and PsbP accumulated efficiently in etioplasts. After 6 h de-etiolation, a complete PSII core complex appeared with 40% of the maximal photochemical efficiency, but a fully functional PSII was recorded only after 24 h illumination. Similarly, the core complex of PSI was assembled after 6 h illumination, whereas the PSI-light-harvesting complex I was stably assembled only in chloroplasts illuminated for 24 h. Moreover, a battery of proteins responsible for defense against oxidative stress accumulated particularly in etioplasts, including the stromal and thylakoidal forms of ascorbate peroxidase, glutathione reductase and PsbS.

  14. Boron Supply Enhances Aluminum Tolerance in Root Border Cells of Pea (Pisum sativum) by Interacting with Cell Wall Pectins.

    PubMed

    Li, Xue Wen; Liu, Jia You; Fang, Jing; Tao, Lin; Shen, Ren Fang; Li, Ya Lin; Xiao, Hong Dong; Feng, Ying Ming; Wen, Hai Xiang; Guan, Jia Hua; Wu, Li Shu; He, Yong Ming; Goldbach, Heiner E; Yu, Min

    2017-01-01

    Aluminum (Al) toxicity is the primary factor limiting crop growth in acidic soils. Boron (B) alleviates Al toxicity in plants, which is mainly considered to be due to the formation of Rhamnogalacturonan II-B (RGII-B) complexes, which helps to stabilize the cytoskeleton. It is unclear yet whether this is due to the increasing of net negative charges and/or further mechanisms. Kinetics of Al accumulation and adsorption were investigated using entire cells, cell wall and pectin of root border cells (RBCs) of pea (Pisum sativum), to reveal the mechanism of B in interacting with alkali-soluble and chelator-soluble pectin for an increased Al tolerance in RBCs. The results show that B could rescue RBCs from Al-induced cell death by accumulating more Al in the cell wall, predominately in alkali-soluble pectin. Boron also promotes Al(3+) adsorption and inhibits Al(3+) desorption from alkali-soluble pectin. Thus, more Al(3+) is immobilized within the alkali-soluble pectin fraction and less in the chelator-soluble pectin, rendering Al(3+) less mobile. Boron induces an increase of RG-II (KDO,2-keto-3-deoxyoctonic acid) content for forming more borate-RGII complexes, and the decrease of pectin methyl-esterification, thus creates more negative charges to immobilize Al(3+) in cell wall pectin. The study provides evidence that abundant B supply enhances the immobilization of Al in alkali-soluble pectin, thus most likely reducing the entry of Al(3+) into the symplast from the surroundings.

  15. Key metabolic traits of Pisum sativum maintain cell vitality during Didymella pinodes infection: cultivar resistance and the microsymbionts' influence.

    PubMed

    Turetschek, Reinhard; Desalegn, Getinet; Epple, Tamara; Kaul, Hans-Peter; Wienkoop, Stefanie

    2017-03-06

    Ascochyta blight causes severe losses in field pea production and the search for resistance traits towards the causal agent Didymella pinodes is of particular importance for farmers. Various microsymbionts have been reported to shape the plants' immune response. However, regardless their contribution to resistance, they are hardly included in experimental designs. We delineate the effect of symbionts (rhizobia, mycorrhiza) on the leaf proteome and metabolome of two field pea cultivars with varying resistance levels against D. pinodes and, furthermore, show cultivar specific symbiont colonisation efficiency. The pathogen infection showed a stronger influence on the interaction with the microsymbionts in the susceptible cultivar, which was reflected in decreased nodule weight and root mycorrhiza colonisation. Vice versa, symbionts induced variation of the host's infection response which, however, was overruled by genotypic resistance associated traits of the tolerant cultivar such as maintenance of photosynthesis and provision of sugars and carbon back bones to fuel secondary metabolism. Moreover, resistance appears to be linked to sulphur metabolism, a functional glutathione-ascorbate hub and fine adjustment of jasmonate and ethylene synthesis to suppress induced cell death. We conclude that these metabolic traits are essential for sustainment of cell vitality and thus, a more efficient infection response. The infection response of two Pisum sativum cultivars with varying resistance levels towards Didymella pinodes was analysed most comprehensively at proteomic and metabolomic levels. Enhanced tolerance was linked to newly discovered cultivar specific metabolic traits such as hormone synthesis and presumably suppression of cell death. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  16. Xyloglucan oligosaccharides promote growth and activate cellulase: Evidence for a role of cellulase in cell expansion. [Pisum sativum L

    SciTech Connect

    McDougall, G.J.; Fry, S.C. )

    1990-07-01

    Oligosaccharides produced by the action of fungal cellulase on xyloglucans promoted the elongation of etiolated pea (Pisum sativum L.) stem segments in a straight-growth bioassay designed for the determination of auxins. The oligosaccharides were most active at about 1 micromolar. We tested the relative growth-promoting activities of four HPLC-purified oligosaccharides which shared a common glucose{sub 4} {center dot} xylose{sub 3} (XG7) core. The substituted oligosaccharides XG8 (glucose{sub 4} {center dot} xylose{sub 3} {center dot} galactose) and XG9n (glucose{sub 4} {center dot} xylose{sub 3} {center dot} galactose{sub 2}) were more effective than XG7 itself and XG9 (glucose{sub 4} {center dot} xylose{sub 3} {center dot} galactose {center dot} fucose). The same oligosaccharides also promoted the degradation, assayed viscometrically, of xyloglucan by an acidic cellulase from bean (Phaseolus vulgaris L.) leaves. The oligosaccharides were highly active at 10{sup {minus}4} molar, causing up to a fourfold increase in activity, but the effect was still detectable at 1 micromolar. Those oligosaccharides (XG8 and XG9n) which best promoted growth, stimulated cellulase activity to the greatest extent. The oligosaccharides did not stimulate the action of the cellulase in an assay based on the conversion of ({sup 3}H)xyloglucan to ethanol-soluble fragments. This suggests that the oligosaccharides enhanced the midchain hydrolysis of xyloglucan molecules (which would rapidly reduce the viscosity of the solution), at the expense of cleavage near the termini (which would yield ethanol-soluble products).

  17. Virus-induced plasma membrane aquaporin PsPIP2;1 silencing inhibits plant water transport of Pisum sativum.

    PubMed

    Song, Juanjuan; Ye, Guoliang; Qian, Zhengjiang; Ye, Qing

    2016-12-01

    Aquaporins (AQPs) are known to facilitate water transport across cell membranes, but the role of a single AQP in regulating plant water transport, particularly in plants other than Arabidopsis remains largely unexplored. In the present study, a virus-induced gene silencing (VIGS) technique was employed to suppress the expression of a specific plasma membrane aquaporin PsPIP2;1 of Pea plants (Pisum sativum), and subsequent effects of the gene suppression on root hydraulic conductivity (Lpr), leaf hydraulic conductivity (K leaf ), root cell hydraulic conductivity (Lprc), and leaf cell hydraulic conductivity (Lplc) were investigated, using hydroponically grown Pea plants. Compared with control plants, VIGS-PsPIP2;1 plants displayed a significant suppression of PsPIP2;1 in both roots and leaves, while the expression of other four PIP isoforms (PsPIP1;1, PsPIP1;2, PsPIP2;2, and PsPIP2;3) that were simultaneously monitored were not altered. As a consequence, significant declines in water transport of VIGS-PsPIP2;1 plants were observed at both organ and cell levels, i.e., as compared to control plants, Lpr and K leaf were reduced by 29 %, and Lprc and Lplc were reduced by 20 and 29 %, respectively. Our results demonstrate that PsPIP2;1 alone contributes substantially to root and leaf water transport in Pea plants, and highlight VIGS a useful tool for investigating the role of a single AQP in regulating plant water transport.

  18. Metabolism of inositol(1,4,5)trisphosphate by a soluble enzyme fraction from pea (Pisum sativum) roots

    SciTech Connect

    Drobak, B.K.; Watkins, P.A.C.; Roberts, K. ); Chattaway, J.A. Univ. of East Anglia, Norwich ); Dawson, A.P. )

    1991-02-01

    Metabolism of the putative messenger molecule D-myo-inositol(1,4,5)trisphosphate (Ins(1,4,5)P{sub 3}) in plant cells has been studied using a soluble fraction from pea (pisum sativum) roots as enzyme source and (5-{sup 32}P)Ins(1,4,5)P{sub 3} and (2-{sup 3}H)Ins(1,4,5)P{sub 3} as tracers. Ins(1,4,5)P{sub 3} was rapidly converted into both lower and higher inositol phosphates. The major dephosphorylation product was inositol (4,5) bisphosphate (Ins(4,5)P{sub 2}) whereas inositol(1,4)bisphosphate (Ins(1,4)P{sub 2}) was only present in very small quantities throughout a 15 minute incubation period. In addition to these compounds, small amounts of nine other metabolites were produced including inositol and inositol(1,4,5,X)P{sub 4}. Dephosphorylation of Ins(1,4,5)P{sub 3} to Ins(4,5)P{sub 2} was dependent on Ins(1,4,5)P{sub 3} concentration and was partially inhibited by the phosphohydrolase inhibitors 2,3-diphosphoglycerate, glucose 6-phosphate, and p-nitrophenylphosphate. Conversion of Ins(1,4,5)P{sub 3} to Ins(4,5)P{sub 2} and Ins(1,4,5,X)P{sub 4} was inhibited by 55 micromolar Ca{sup 2+}. This study demonstrates that enzymes are present in plant tissues which are capable of rapidly converting Ins(1,4,5)P{sub 3} and that pathways of inositol phosphate metabolism exist which may prove to be unique to the plant kingdom.

  19. Boron Supply Enhances Aluminum Tolerance in Root Border Cells of Pea (Pisum sativum) by Interacting with Cell Wall Pectins

    PubMed Central

    Fang, Jing; Tao, Lin; Shen, Ren Fang; Li, Ya Lin; Xiao, Hong Dong; Feng, Ying Ming; Wen, Hai Xiang; Guan, Jia Hua; Wu, Li Shu; He, Yong Ming; Goldbach, Heiner E.; Yu, Min

    2017-01-01

    Aluminum (Al) toxicity is the primary factor limiting crop growth in acidic soils. Boron (B) alleviates Al toxicity in plants, which is mainly considered to be due to the formation of Rhamnogalacturonan II-B (RGII-B) complexes, which helps to stabilize the cytoskeleton. It is unclear yet whether this is due to the increasing of net negative charges and/or further mechanisms. Kinetics of Al accumulation and adsorption were investigated using entire cells, cell wall and pectin of root border cells (RBCs) of pea (Pisum sativum), to reveal the mechanism of B in interacting with alkali-soluble and chelator-soluble pectin for an increased Al tolerance in RBCs. The results show that B could rescue RBCs from Al-induced cell death by accumulating more Al in the cell wall, predominately in alkali-soluble pectin. Boron also promotes Al3+ adsorption and inhibits Al3+ desorption from alkali-soluble pectin. Thus, more Al3+ is immobilized within the alkali-soluble pectin fraction and less in the chelator-soluble pectin, rendering Al3+ less mobile. Boron induces an increase of RG-II (KDO,2-keto-3-deoxyoctonic acid) content for forming more borate-RGII complexes, and the decrease of pectin methyl-esterification, thus creates more negative charges to immobilize Al3+ in cell wall pectin. The study provides evidence that abundant B supply enhances the immobilization of Al in alkali-soluble pectin, thus most likely reducing the entry of Al3+ into the symplast from the surroundings. PMID:28533794

  20. Pisum sativum p68 DEAD-box protein is ATP-dependent RNA helicase and unique bipolar DNA helicase.

    PubMed

    Tuteja, Narendra; Tarique, Mohammed; Banu, Mst Sufara Akhter; Ahmad, Moaz; Tuteja, Renu

    2014-08-01

    DEAD-box helicases play essential role in DNA and RNA metabolism such as replication, repair, recombination, transcription, translation, ribosome biogenesis and splicing which regulate plant growth and development. The presence of helicases in the stress-induced ORFs identified by cDNA microarray indicates that helicases might be playing an important role in stabilizing growth in plants under stress. p68 DEAD-box helicase has been identified and characterized from animal systems but the properties and functions of plant p68 are poorly understood. In this study, the identification, purification and characterization of recombinant p68 from Pisum sativum (Psp68) is presented. Psp68 possesses all the characteristic motifs like DEAD-box ATP-binding and helicase C terminal motifs and is structurally similar to human p68 homologue. Psp68 exhibits ATPase activity in the presence of both DNA and RNA and it binds to DNA as well as RNA. It contains the characteristic RNA helicase activity. Interestingly Psp68 also shows the unique DNA helicase activity, which is bipolar in nature (unwinds DNA in both the 5'-3' and 3'-5' directions). The Km values of Psp68 for ATPase are 0.5126 and 0.9142 mM in the presence of DNA and RNA, respectively. The Km values of Psp68 are 1.6129 and 1.14 nM for DNA helicase and RNA helicase, respectively. The unique properties of Psp68 suggest that it could be a multifunctional protein involved in different aspect of DNA and RNA metabolism. This discovery should make an important contribution to better understanding of nucleic acids metabolism plants.

  1. Expression of arginine decarboxylase is induced during early fruit development and in young tissues of Pisum sativum (L.).

    PubMed

    Pérez-Amador, M A; Carbonell, J; Granell, A

    1995-09-01

    A cDNA coding for arginine decarboxylase (ADC, EC 4.1.1.19) has been isolated from a cDNA library of parthenocarpic young fruits of Pisum sativum (L.). The deduced aminoacid sequence is 74%, 46% and 35% identical to ADCs from tomato, oat and Escherichia coli, respectively. When the pea ADC cDNA was put under the control of the galactose inducible yeast promoter CYC1-GAL10 and introduced into Saccharomyces cerevisiae, it conferred galactose-regulated expression of the ADC activity. The ADC activity expressed in S. cerevisiae was inhibited 99% by alpha-DL-difluoromethylarginine (DFMA), a specific inhibitor of ADC activity. No activity was detected in the untransformed S. cerevisiae, nor when it was transformed with an antisense ADC construct. This provides direct evidence that the ADC cDNA from pea encoded a functional, specific ADC activity and that S. cerevisiae is able to process correctly the protein. In the pea plant, gene expression of the ADC is high in young developing tissues like shoot tips, young leaflets and flower buds. Fully expanded leaflets and roots have much lower, but still detectable, levels of the ADC transcript. In the ovary and fruit, they are developmentally regulated, showing high levels of expression during the early stages of fruit growth, which in pea is mainly due to cell expansion. The observed changes in the steady-state levels of ADC mRNA alone, however, cannot account for the differences in ADC activity suggesting that other regulatory mechanisms must be acting.

  2. A DNA helicase from Pisum sativum is homologous to translation initiation factor and stimulates topoisomerase I activity.

    PubMed

    Pham, X H; Reddy, M K; Ehtesham, N Z; Matta, B; Tuteja, N

    2000-10-01

    DNA helicases play an essential role in all aspects of nucleic acid metabolism, by providing a duplex-unwinding function. This is the first report of the isolation of a cDNA (1.6 kb) clone encoding functional DNA helicase from a plant (pea, Pisum sativum). The deduced amino-acid sequence has eight conserved helicase motifs of the DEAD-box protein family. It is a unique member of this family, containing DESD and SRT motifs instead of DEAD/H and SAT. The encoded 45.5 kDa protein has been overexpressed in bacteria and purified to homogeneity. The purified protein contains ATP-dependent DNA and RNA helicase, DNA-dependent ATPase, and ATP-binding activities. The protein sequence contains striking homology with eIF-4A, which has not so far been reported as DNA helicase. The antibodies against pea helicase inhibit in vitro translation. The gene is expressed as 1.6 kb mRNA in different organs of pea. The enzyme is localized in the nucleus and cytosol, and unwinds DNA in the 3' to 5' direction. The pea helicase interacts with pea topoisomerase I protein and stimulates its activity. These results suggest that pea DNA helicase could be an important multifunctional protein involved in protein synthesis, maintaining the basic activities of the cell, and in upregulation of topoisomerase I activity. The discovery of such a protein with intrinsic multiple activity should make an important contribution to our better understanding of DNA and RNA transactions in plants.

  3. Cadmium accumulation and buffering of cadmium-induced stress by arbuscular mycorrhiza in three Pisum sativum L. genotypes.

    PubMed

    Rivera-Becerril, Facundo; Calantzis, Catherine; Turnau, Katarzyna; Caussanel, Jean-Pierre; Belimov, Andrei A; Gianinazzi, Silvio; Strasser, Reto J; Gianinazzi-Pearson, Vivienne

    2002-05-01

    The role of arbuscular mycorrhiza in reducing Cd stress was investigated in three genotypes of Pisum sativum L. (cv. Frisson, VIR4788, VIR7128), grown in soil/sand pot cultures in the presence and absence of 2-3 mg kg(-1) bioavailable Cd, and inoculated or not with the arbuscular mycorrhizal fungus Glomus intraradices. Shoot, root and pod biomass were decreased by Cd in non-mycorrhizal plants. The presence of mycorrhiza attenuated the negative effect of Cd so that shoot biomass and activity of photosystem II, based on chlorophyll a fluorescence, were not significantly different between mycorrhizal plants growing in the presence or absence of the heavy metal (HM). Total P concentrations were not significantly different between mycorrhizal and non-mycorrhizal plants treated with Cd. From 20-50-fold more Cd accumulated in roots than in shoots of Cd-treated plants, and overall levels were comparable to other metal-accumulating plants. Genetic variability in Cd accumulation existed between the pea genotypes. Concentration of the HM was lowest in roots of VIR4788 and in pods of VIR4788 and VIR7128. G. intraradices inoculation decreased Cd accumulation in roots and pods of cv. Frisson, whilst high concentrations were maintained in roots and pods of mycorrhizal VIR7128. Shoot concentrations of Cd increased in mycorrhizal cv. Frisson and VIR4788. Sequestration of Cd in root cell walls and/or cytoplasm, measured by EDS/SEM, was comparable between non-mycorrhizal pea genotypes but considerably decreased in mycorrhizal cv. Frisson and VIR7128. Possible mechanisms for mycorrhiza buffering of Cd-induced stress in the pea genotypes are discussed.

  4. Photosynthetic carbon metabolism in leaflets, stipules and tendrils of Pisum sativum L

    SciTech Connect

    Cote, R.; Grodzinski, B. )

    1990-05-01

    Gas exchange and photosynthetic carbon metabolism have been investigated for each of the dominant parts of the pea leaf (P. sativum) in a normal and a semi-leafless phenotype (cv. Improved Laxton's Progress, and cv. Curly, respectively). On a fresh weight basis, net photosynthesis of leaflets and stipules have similar rates, while in tendrils the rte is 40% lower. However, on a surface area basis, tendrils are only 5-10% less efficient photosynthetically when the area is corrected by a factor {pi}/2. Transpiration rates are similar for leaflets and stipules, but double for tendrils even though stomatal frequency on tendrils is reduced by 50%. Dark respiration is higher in tendrils than leaflets and stipules. Gas exchange is comparable in both cultivars. The early {sup 14}C-labelled products of stipules, leaflets and tendrils are similar in both phenotypes, however the tendrils clearly partition about 2-3 times more of the newly fixed {sup 14}CO{sub 2} into the amino acid fraction. These data will be discussed in relation to the anatomy and function of pea tendrils.

  5. High-Throughput Development of SSR Markers from Pea (Pisum sativum L.) Based on Next Generation Sequencing of a Purified Chinese Commercial Variety

    PubMed Central

    Zhang, Xiaoyan; Hu, Jinguo; Bao, Shiying; Hao, Junjie; Li, Ling; He, Yuhua; Jiang, Junye; Wang, Fang; Tian, Shufang; Zong, Xuxiao

    2015-01-01

    Pea (Pisum sativum L.) is an important food legume globally, and is the plant species that J.G. Mendel used to lay the foundation of modern genetics. However, genomics resources of pea are limited comparing to other crop species. Application of marker assisted selection (MAS) in pea breeding has lagged behind many other crops. Development of a large number of novel and reliable SSR (simple sequence repeat) or microsatellite markers will help both basic and applied genomics research of this crop. The Illumina HiSeq 2500 System was used to uncover 8,899 putative SSR containing sequences, and 3,275 non-redundant primers were designed to amplify these SSRs. Among the 1,644 SSRs that were randomly selected for primer validation, 841 yielded reliable amplifications of detectable polymorphisms among 24 genotypes of cultivated pea (Pisum sativum L.) and wild relatives (P. fulvum Sm.) originated from diverse geographical locations. The dataset indicated that the allele number per locus ranged from 2 to 10, and that the polymorphism information content (PIC) ranged from 0.08 to 0.82 with an average of 0.38. These 1,644 novel SSR markers were also tested for polymorphism between genotypes G0003973 and G0005527. Finally, 33 polymorphic SSR markers were anchored on the genetic linkage map of G0003973 × G0005527 F2 population. PMID:26440522

  6. High-Throughput Development of SSR Markers from Pea (Pisum sativum L.) Based on Next Generation Sequencing of a Purified Chinese Commercial Variety.

    PubMed

    Yang, Tao; Fang, Li; Zhang, Xiaoyan; Hu, Jinguo; Bao, Shiying; Hao, Junjie; Li, Ling; He, Yuhua; Jiang, Junye; Wang, Fang; Tian, Shufang; Zong, Xuxiao

    2015-01-01

    Pea (Pisum sativum L.) is an important food legume globally, and is the plant species that J.G. Mendel used to lay the foundation of modern genetics. However, genomics resources of pea are limited comparing to other crop species. Application of marker assisted selection (MAS) in pea breeding has lagged behind many other crops. Development of a large number of novel and reliable SSR (simple sequence repeat) or microsatellite markers will help both basic and applied genomics research of this crop. The Illumina HiSeq 2500 System was used to uncover 8,899 putative SSR containing sequences, and 3,275 non-redundant primers were designed to amplify these SSRs. Among the 1,644 SSRs that were randomly selected for primer validation, 841 yielded reliable amplifications of detectable polymorphisms among 24 genotypes of cultivated pea (Pisum sativum L.) and wild relatives (P. fulvum Sm.) originated from diverse geographical locations. The dataset indicated that the allele number per locus ranged from 2 to 10, and that the polymorphism information content (PIC) ranged from 0.08 to 0.82 with an average of 0.38. These 1,644 novel SSR markers were also tested for polymorphism between genotypes G0003973 and G0005527. Finally, 33 polymorphic SSR markers were anchored on the genetic linkage map of G0003973 × G0005527 F2 population.

  7. Nitric oxide alleviates silver nanoparticles (AgNps)-induced phytotoxicity in Pisum sativum seedlings.

    PubMed

    Tripathi, Durgesh Kumar; Singh, Swati; Singh, Shweta; Srivastava, Prabhat Kumar; Singh, Vijay Pratap; Singh, Samiksha; Prasad, Sheo Mohan; Singh, Prashant Kumar; Dubey, Nawal Kishore; Pandey, Avinash Chand; Chauhan, Devendra Kumar

    2017-01-01

    Understanding the adverse impact of nanoparticles in crop plants has emerged as one of the most interesting fields of plant research. Therefore, this study has been conducted to investigate the impact of silver nanoparticles (AgNps) on Pisium sativum seedlings. Besides this, we have also tested whether nitric oxide (NO) is capable of reducing toxicity of AgNps or not. NO has been found as one of the most fascinating molecules, capable of enhancing plant tolerance to different environmental stresses. The results of the present study showed that AgNps treatments (1000 μM and 3000 μM) significantly declined growth parameters, photosynthetic pigments and chlorophyll fluorescence of pea seedlings, which could be correlated with increased accumulation of Ag in root and shoot of pea seedlings. In contrast, addition of SNP (100 μM; a donor of NO) successfully ameliorated AgNp-induced adverse effects on these parameters as it reduced accumulation of Ag and repaired damaged tissues. Levels of oxidative stress markers (SOR, H2O2 and MDA) were enhanced while their levels significantly reduced under SNP addition. AgNps (1000 μM and 3000 μM) significantly stimulated the activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX) while inhibited activities of glutathione reductase (GR) and dehydroascorbate reductase (DHAR). AgNps also considerably declined the total ascorbate and glutathione contents and severely damaged leaf and root anatomical structures. On the other hand, addition of SNP further increased the level of SOD, APX, GR and DHAR and significantly increased the decreased levels of total ascorbate and glutathione contents, and repaired anatomical structures. In conclusion, this study suggests that AgNps treatments adversely decreased growth, pigments and photosynthesis due to enhanced level of Ag and oxidative stress. However, SNP addition successfully ameliorates adverse impact of AgNps on pea seedlings by regulating the Ag uptake, antioxidant

  8. Interaction between cochleata and stipule-reduced mutations results in exstipulate hypertrophied leaves in Pisum sativum L.

    PubMed

    Kumar, Arvind; Sharma, Vishakha; Kumar, Sushil

    2013-07-01

    In the wild type P. sativum, each of the adult plant stem nodes, bears a pair of sessile foliaceous stipules and a petiolated unipinnately compound leaf of 4 to 6 leaflets and 7-9 tendrils. The stipule-reduced (st) and cochleata (coch) single null mutants and coch st double null mutant differ fom the wild type in respectively having sessile stipules of much reduced size, petiolated simple and/or compound leaf-like stipules and no stipules. It is also known that coch leaves are somewhat bigger than st and wild type leaves. Here, pleiotropic phenotype of coch st double mutant was investigated. The morphologies of stipules and leaf were quantified in the field grown plants and microcultured shoots, latter in the presence and absence of gibberellic acid and N-1-naphthylphthalamic acid. The observations showed that as compared to the corresponding plants or shoots of COCH ST (WT) genotype, (a) coch st plants bore leaves in which all the organs were hypertrophied; (b) full complement of leaflets and 3-5 tendrils were formed on leaf; (c) the microcultured coch st shoots were taller despite lower number of nodes, and (d) they also produced leaves in which all the organs were bigger and the ratio of leaflets/tendrils was higher. It was concluded that in coch st double mutant (a) ST function is essential for stipule primordium differentiation, in the absence of COCH function and (b) absence of negative feedback loops between simple stipules and compound leaf for metabolite utilization allows hypertrophied growth in leaves.

  9. Recovery from Photoinhibition in Peas (Pisum sativum L.) Acclimated to Varying Growth Irradiances (Role of D1 Protein Turnover).

    PubMed Central

    Aro, E. M.; McCaffery, S.; Anderson, J. M.

    1994-01-01

    D1 protein turnover and restoration of the photochemical efficiency of photosystem II (PSII) after photoinhibition of pea leaves (Pisum sativum L. cv Greenfeast) acclimated to different light intensities were investigated. All peas acclimated to different light intensities were able to recover from photoinhibition, at least partially, at light intensities far above their growth light irradiance. However, the capacity of pea leaves to recover from photoinhibition under increasing high irradiances was strictly dependent on the light acclimation of the leaves; i.e. the higher the irradiance during growth, the better the capacity of pea leaves to recover from photoinhibition at moderate and high light. In our experimental conditions, mainly D1 protein turnover-dependent recovery was monitored, since in the presence of an inhibitor of chloroplast-encoded protein synthesis, lincomycin, only negligible recovery took place. In darkness, neither the restoration of PSII photochemical efficiency nor any notable degradation of damaged D1 protein took place. In low light, however, good recovery of PSII occurred in all peas acclimated to different light intensities and was accompanied by fast degradation of the D1 protein. The rate of degradation of the D1 protein was estimated to be 3 to 4 times faster in photoinhibited leaves than in nonphotoinhibited leaves under the recovery conditions of 50 [mu]mol of photons m-2 s-1. In moderate light of 400 [mu]mol of photons m-2 s-1, the photoinhibited low-light peas were not able to increase further the rate of D1 protein degradation above that observed in nonphotoinhibited leaves, nor was the restoration of PSII function possible. On the other hand, photoinhibited high-light leaves were able to increase the rate of D1 protein degradation above that of nonphotoinhibited leaves even in moderate and high light, ensuring at least partial restoration of PSII function. We conclude that the capacity of photoinhibited leaves to restore PSII

  10. Photosynthesis light-independent reactions are sensitive biomarkers to monitor lead phytotoxicity in a Pb-tolerant Pisum sativum cultivar.

    PubMed

    Rodriguez, Eleazar; da Conceição Santos, Maria; Azevedo, Raquel; Correia, Carlos; Moutinho-Pereira, José; Ferreira de Oliveira, José Miguel Pimenta; Dias, Maria Celeste

    2015-01-01

    Lead (Pb) environmental contamination remains prevalent. Pisum sativum L. plants have been used in ecotoxicological studies, but some cultivars showed to tolerate and accumulate some levels of Pb, opening new perspectives to their use in phytoremediation approaches. However, the putative use of pea plants in phytoremediation requires reliable toxicity endpoints. Here, we evaluated the sensitivity of a large number of photosynthesis-related biomarkers in Pb-exposed pea plants. Plants (cv. "Corne de Bélier") were exposed to Pb concentrations up to 1,000 mg kg(-1) soil during 28 days. The photosynthetic potential biomarkers that were analyzed included pigments, chlorophyll (Chl) a fluorescence, gas exchange, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) activity, and carbohydrates. Flow cytometry (FCM) was also used to assess the morpho-functional status of chloroplasts. Finally, Pb-induced nutrient disorders were also evaluated. Net CO2 assimilation rate (A) and RuBisCO activity decreased strongly in Pb-exposed plants. Plant dry mass (DM) accumulation, however, was only reduced in the higher Pb concentrations tested (500 and 1,000 mg kg(-1) soil). Pigment contents increased solely in plants exposed to the largest Pb concentration, and in addition, the parameters related to the light-dependent reactions of photosynthesis, Fv/Fm and ΦPSII, were not affected by Pb exposure. In contrast to this, carbohydrates showed an overall tendency to increase in Pb-exposed plants. The morphological status of chloroplasts was affected by Pb exposure, with a general trend of volume decrease and granularity increase. These results point the endpoints related to the light-independent reactions of photosynthesis as more sensitive predictors of Pb-toxicity than the light-dependent reactions ones. Among the endpoints related to the light-independent photosynthesis reactions, RuBisCO activity and A were found to be the most sensitive. We discuss here the advantages of using

  11. Solvent hydrogen isotope effects and anion inhibition of CO2 hydration catalysed by carbonic anhydrase from Pisum sativum.

    PubMed

    Johansson, I M; Forsman, C

    1994-09-15

    Chloroplast carbonic anhydrase from Pisum sativum has been studied to elucidate the catalytic mechanism and to test if the mechanism proposed for human carbonic anhydrase II is also valid for pea carbonic anhydrase. The catalytic activity was found to depend on the chemical nature of the buffer. Barbital buffer gives the highest turnover number at infinite buffer concentration and the lowest Km value with respect to the buffer, while the kinetic parameters obtained in the imidazole-type buffer, 1-methylimidazole, do not differ from those obtained using the biological-type buffer Mops. The anion inhibition of CO2 hydration was investigated using SCN- at pH 6-9. The binding of the anion was found to be pH dependent with the strongest interaction at low pH. We obtained an uncompetitive inhibition pattern at high pH and noncompetitive inhibition patterns at pH 7 and low pH. The catalytic mechanism was further tested by measurements of the solvent hydrogen isotope effects on the kinetic parameters for CO2 hydration. The observed effects were comparatively small with a kcat value of approximately 2 irrespective of the pH. The effect on kcat/Km and on Km changes when going from high pH to pH 7 and low pH. At high pH, the solvent isotope effect in Km is at least 3, giving a value below 1 for kcat/Km, while at pH 7 and low pH the major effect is found in kcat/Km with values of 2.6 and 2.9. The dependence of the CO2-hydration activity on the buffer concentration is in agreement with a ping-pong mechanism with buffer acting as a second substrate. This is analogous to the behaviour of human carbonic anhydrase II. The inhibition patterns and the observed isotope effects at high pH can also be explained within the framework of the catalytic mechanism for human carbonic anhydrase II, with a rate-determining and buffer-dependent part. The results are consistent with a mechanism involving a proton transfer that contributes to rate limitation. However, the isotope effects found at p

  12. SNP marker discovery, linkage map construction and identification of QTLs for enhanced salinity tolerance in field pea (Pisum sativum L.)

    PubMed Central

    2013-01-01

    Background Field pea (Pisum sativum L.) is a self-pollinating, diploid, cool-season food legume. Crop production is constrained by multiple biotic and abiotic stress factors, including salinity, that cause reduced growth and yield. Recent advances in genomics have permitted the development of low-cost high-throughput genotyping systems, allowing the construction of saturated genetic linkage maps for identification of quantitative trait loci (QTLs) associated with traits of interest. Genetic markers in close linkage with the relevant genomic regions may then be implemented in varietal improvement programs. Results In this study, single nucleotide polymorphism (SNP) markers associated with expressed sequence tags (ESTs) were developed and used to generate comprehensive linkage maps for field pea. From a set of 36,188 variant nucleotide positions detected through in silico analysis, 768 were selected for genotyping of a recombinant inbred line (RIL) population. A total of 705 SNPs (91.7%) successfully detected segregating polymorphisms. In addition to SNPs, genomic and EST-derived simple sequence repeats (SSRs) were assigned to the genetic map in order to obtain an evenly distributed genome-wide coverage. Sequences associated with the mapped molecular markers were used for comparative genomic analysis with other legume species. Higher levels of conserved synteny were observed with the genomes of Medicago truncatula Gaertn. and chickpea (Cicer arietinum L.) than with soybean (Glycine max [L.] Merr.), Lotus japonicus L. and pigeon pea (Cajanus cajan [L.] Millsp.). Parents and RIL progeny were screened at the seedling growth stage for responses to salinity stress, imposed by addition of NaCl in the watering solution at a concentration of 18 dS m-1. Salinity-induced symptoms showed normal distribution, and the severity of the symptoms increased over time. QTLs for salinity tolerance were identified on linkage groups Ps III and VII, with flanking SNP markers suitable for

  13. Discovery of a Novel er1 Allele Conferring Powdery Mildew Resistance in Chinese Pea (Pisum sativum L.) Landraces

    PubMed Central

    Sun, Suli; Fu, Haining; Wang, Zhongyi; Duan, Canxing; Zong, Xuxiao; Zhu, Zhendong

    2016-01-01

    Pea powdery mildew, caused by Erysiphe pisi D.C., is an important disease worldwide. Deployment of resistant varieties is the main way to control this disease. This study aimed to screen Chinese pea (Pisum sativum L.) landraces resistant to E. pisi, and to characterize the resistance gene(s) at the er1 locus in the resistant landraces, and to develop functional marker(s) specific to the novel er1 allele. The 322 landraces showed different resistance levels. Among them, 12 (3.73%), 4 (1.24%) and 17 (5.28%) landraces showed immunity, high resistance and resistance to E. pisi, respectively. The other landraces appeared susceptible or highly susceptible to E. pisi. Most of the immune and highly resistant landraces were collected from Yunnan province. To characterize the resistance gene at the er1 locus, cDNA sequences of PsMLO1 gene were determined in 12 immune and four highly resistant accessions. The cDNAs of PsMLO1 from the immune landrace G0005576 produced three distinct transcripts, characterized by a 129-bp deletion, and 155-bp and 220-bp insertions, which were consistent with those of er1-2 allele. The PsMLO1 cDNAs in the other 15 resistant landraces produced identical transcripts, which had a new point mutation (T→C) at position 1121 of PsMLO1, indicating a novel er1 allele, designated as er1-6. This mutation caused a leucine to proline change in the amino acid sequence. Subsequently, the resistance allele er1-6 in landrace G0001778 was confirmed by resistance inheritance analysis and genetic mapping on the region of the er1 locus using populations derived from G0001778 × Bawan 6. Finally, a functional marker specific to er1-6, SNP1121, was developed using the high-resolution melting technique, which could be used in pea breeding via marker-assisted selection. The results described here provide valuable genetic information for Chinese pea landraces and a powerful tool for pea breeders. PMID:26809053

  14. Discovery of a Novel er1 Allele Conferring Powdery Mildew Resistance in Chinese Pea (Pisum sativum L.) Landraces.

    PubMed

    Sun, Suli; Fu, Haining; Wang, Zhongyi; Duan, Canxing; Zong, Xuxiao; Zhu, Zhendong

    2016-01-01

    Pea powdery mildew, caused by Erysiphe pisi D.C., is an important disease worldwide. Deployment of resistant varieties is the main way to control this disease. This study aimed to screen Chinese pea (Pisum sativum L.) landraces resistant to E. pisi, and to characterize the resistance gene(s) at the er1 locus in the resistant landraces, and to develop functional marker(s) specific to the novel er1 allele. The 322 landraces showed different resistance levels. Among them, 12 (3.73%), 4 (1.24%) and 17 (5.28%) landraces showed immunity, high resistance and resistance to E. pisi, respectively. The other landraces appeared susceptible or highly susceptible to E. pisi. Most of the immune and highly resistant landraces were collected from Yunnan province. To characterize the resistance gene at the er1 locus, cDNA sequences of PsMLO1 gene were determined in 12 immune and four highly resistant accessions. The cDNAs of PsMLO1 from the immune landrace G0005576 produced three distinct transcripts, characterized by a 129-bp deletion, and 155-bp and 220-bp insertions, which were consistent with those of er1-2 allele. The PsMLO1 cDNAs in the other 15 resistant landraces produced identical transcripts, which had a new point mutation (T→C) at position 1121 of PsMLO1, indicating a novel er1 allele, designated as er1-6. This mutation caused a leucine to proline change in the amino acid sequence. Subsequently, the resistance allele er1-6 in landrace G0001778 was confirmed by resistance inheritance analysis and genetic mapping on the region of the er1 locus using populations derived from G0001778 × Bawan 6. Finally, a functional marker specific to er1-6, SNP1121, was developed using the high-resolution melting technique, which could be used in pea breeding via marker-assisted selection. The results described here provide valuable genetic information for Chinese pea landraces and a powerful tool for pea breeders.

  15. Modulation of CuO nanoparticles toxicity to green pea (Pisum sativum Fabaceae) by the phytohormone indole-3-acetic acid.

    PubMed

    Ochoa, Loren; Medina-Velo, Illya A; Barrios, Ana C; Bonilla-Bird, Nestor J; Hernandez-Viezcas, Jose A; Peralta-Videa, Jose R; Gardea-Torresdey, Jorge L

    2017-11-15

    The response of plants to copper oxide nanoparticles (nano-CuO) in presence of exogenous phytohormones is unknown. In this study, green pea (Pisum sativum) plants were cultivated to full maturity in soil amended with nano-CuO (10-100nm, 74.3% Cu), bulk-CuO (bCuO, 100-10,000nm, 79.7% Cu), and CuCl2 at 50 and 100mg/kg and indole-3-acetic acid (IAA) at 10 and 100μM. Results showed that IAA at 10 and 100μM, averaged over all Cu treatments, reduced the number of plants by ~23% and ~34%, respectively. IAA at 10μM, nano-CuO at 50mg/kg, b-CuO at 50mg/kg, and CuCl2 at 100mg/kg reduced pod biomass by about 50%. Although some combinations of IAA, mainly at 100μM, with the Cu compounds altered nutrient accumulation in tissues, none of them affected pod elements. Conversely, without IAA, nano-CuO at 50mg/kg, increased pod Fe and Ni by 258% and 325%, respectively, while bCuO at 100mg/kg increased pod Ni by 275%, compared with control. With IAA at 10μM, nano-CuO (100mg/kg) and bCuO (50mg/kg) increased stem Cu by ~84% and ~78%. When IAA increased to 100μM, nano-CuO and bCuO reduced stem Ca by 32% and 37%, and Mg by ~35%. Results suggest that both the nano-CuO and bCuO could improve the nutritional quality of pea pods, while exogenous IAA combined with Cu-based compounds could impact green pea production since these treatments reduced the number of plants and pod biomass. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Differential Toxicity of Bare and Hybrid ZnO Nanoparticles in Green Pea (Pisum sativum L.): A Life Cycle Study.

    PubMed

    Mukherjee, Arnab; Sun, Youping; Morelius, Erving; Tamez, Carlos; Bandyopadhyay, Susmita; Niu, Genhua; White, Jason C; Peralta-Videa, Jose R; Gardea-Torresdey, Jorge L

    2015-01-01

    The effect of surface or lattice modification of nanoparticles (NPs) on terrestrial plants is poorly understood. We investigated the impact of different zinc oxide (ZnO) NPs on green pea (Pisum sativum L.), one of the highest consumed legumes globally. Pea plants were grown for 65 d in soil amended with commercially available bare ZnO NPs (10 nm), 2 wt% alumina doped (Al2O3@ZnO NPs, 15 nm), or 1 wt% aminopropyltriethoxysilane coated NPs (KH550@ZnO NP, 20 nm) at 250 and 1000 mg NP/kg soil inside a greenhouse. Bulk (ZnO) and ionic Zn (zinc chloride) were included as controls. Plant fresh and dry biomass, changes in leaf pigment concentrations, elements (Zn, Al, Si), and protein and carbohydrate profile of green pees were quantified upon harvest at 65 days. With the exception of the coated 1000 mg/kg NP treatment, fresh and dry weight were unaffected by Zn exposure. Although, all treated plants showed higher tissue Zn than controls, those exposed to Al2O3@ZnO NPs at 1000 mg/kg had greater Zn concentration in roots and seeds, compared to bulk Zn and the other NP treatments, keeping Al and Si uptake largely unaffected. Higher Zn accumulation in green pea seeds were resulted in coated ZnO at 250 mg/kg treatments. In leaves, Al2O3@ZnO NP at 250 mg/kg significantly increased Chl-a and carotenoid concentrations relative to the bulk, ionic, and the other NP treatments. The protein and carbohydrate profiles remained largely unaltered across all treatments with the exception of Al2O3@ZnO NPs at 1000 mg/kg where sucrose concentration of green peas increased significantly, which is likely a biomarker of stress. Importantly, these findings demonstrate that lattice and surface modification can significantly alter the fate and phytotoxic effects of ZnO NPs in food crops and seed nutritional quality. To the authors' knowledge, this is the first report of a life cycle study on comparative toxicity of bare, coated, and doped ZnO NPs on a soil-grown food crop.

  17. Differential Toxicity of Bare and Hybrid ZnO Nanoparticles in Green Pea (Pisum sativum L.): A Life Cycle Study

    PubMed Central

    Mukherjee, Arnab; Sun, Youping; Morelius, Erving; Tamez, Carlos; Bandyopadhyay, Susmita; Niu, Genhua; White, Jason C.; Peralta-Videa, Jose R.; Gardea-Torresdey, Jorge L.

    2016-01-01

    The effect of surface or lattice modification of nanoparticles (NPs) on terrestrial plants is poorly understood. We investigated the impact of different zinc oxide (ZnO) NPs on green pea (Pisum sativum L.), one of the highest consumed legumes globally. Pea plants were grown for 65 d in soil amended with commercially available bare ZnO NPs (10 nm), 2 wt% alumina doped (Al2O3@ZnO NPs, 15 nm), or 1 wt% aminopropyltriethoxysilane coated NPs (KH550@ZnO NP, 20 nm) at 250 and 1000 mg NP/kg soil inside a greenhouse. Bulk (ZnO) and ionic Zn (zinc chloride) were included as controls. Plant fresh and dry biomass, changes in leaf pigment concentrations, elements (Zn, Al, Si), and protein and carbohydrate profile of green pees were quantified upon harvest at 65 days. With the exception of the coated 1000 mg/kg NP treatment, fresh and dry weight were unaffected by Zn exposure. Although, all treated plants showed higher tissue Zn than controls, those exposed to Al2O3@ZnO NPs at 1000 mg/kg had greater Zn concentration in roots and seeds, compared to bulk Zn and the other NP treatments, keeping Al and Si uptake largely unaffected. Higher Zn accumulation in green pea seeds were resulted in coated ZnO at 250 mg/kg treatments. In leaves, Al2O3@ZnO NP at 250 mg/kg significantly increased Chl-a and carotenoid concentrations relative to the bulk, ionic, and the other NP treatments. The protein and carbohydrate profiles remained largely unaltered across all treatments with the exception of Al2O3@ZnO NPs at 1000 mg/kg where sucrose concentration of green peas increased significantly, which is likely a biomarker of stress. Importantly, these findings demonstrate that lattice and surface modification can significantly alter the fate and phytotoxic effects of ZnO NPs in food crops and seed nutritional quality. To the authors' knowledge, this is the first report of a life cycle study on comparative toxicity of bare, coated, and doped ZnO NPs on a soil-grown food crop. PMID:26793219

  18. The lld mutation in Pisum sativum used as a genetic tool to discern the plant leaflet/leaf developmental process.

    PubMed

    Sharma, Vishakha; Tripathi, Bhumi Nath; Kumar, Sushil

    2013-06-01

    Leaves of P. sativum the double mutant genotype tendril-less (tl) leaflet-development (lld), due to the action of lld mutation, produce many leaflets that are aborted at different stages of development. Morphological, vein pattern and histological observations showed that aborted leaflets became cup/bell/trumpet (cup) shaped because of segmental differentiation in the leaflet primordium. Cup's inside lamina surface was adaxial and outer surfaces of cup and its stem were abaxial. The lld cups were phenotypically homologous to aborted leaves described in Arabidopsis thaliana mutants, angustifolia and those which underexpressed the HD-ZIP III proteins. Leaflet primordium was found to grow and establish three dimensional polarities apex-downwards. Primordium produced lateral outgrowth on one side of midvein. Differentiation, in the outgrowth of secondary veins, whose xylem tissues faced each other, established the adaxial-abaxial polarity. Lateral outgrowth then developed a cavity which got bounded by future adaxial epidermis. Further growth, veinlet formation, differentiation of palisade parenchyma and spongy parenchyma followed. Opening of lateral outgrowth at its outer midline produced a flat leaflet with lateral lamina spans. The structural and functional correspondence between leaflet and simple leaves suggested commonality between leaf and leaflet development mechanisms. A molecular model for the lld led leaflet abortion was also provided.

  19. Development and Partial Characterization of Nearly Isogenic Pea Lines (Pisum sativum L.) that Alter Uptake Hydrogenase Activity in Symbiotic Rhizobium1

    PubMed Central

    Phillips, Donald A.; Kapulnik, Yoram; Bedmar, Eulogio J.; Joseph, Cecillia M.

    1990-01-01

    Some Rhizobium bacteria have H2-uptake (Hup) systems that oxidize H2 evolved from nitrogenase in leguminous root nodules. Pea (Pisum sativum L.) cultivars `JI1205' and `Alaska' produce high Hup (Hup++) and moderate Hup (Hup+) phenotypes, respectively, in Rhizobium leguminosarum 128C53. The physiological significance and biochemical basis of this host plant genetic effect are unknown. The purpose of this investigation was to advance basic Hup studies by developing nearly isogenic lines of peas that alter Hup phenotypes in R. leguminosarum strains containing hup genes. Eight pairs of nearly isogenic pea lines that produce Hup++ and Hup+ phenotypes in R. leguminosarum 128C53 were identified in 173 F2-derived F6 families produced from crosses between JI1205 and Alaska. Tests with the pea isolines and three strains of hup-containing R. leguminosarum showed that the isolines altered Hup activity significantly (P ≤ 0.05) in 19 of 24 symbiotic combinations. Analyses of Hup phenotypes in F6 families, the F1 population, and two backcrosses suggested involvement of a single genetic locus. Three of the eight pairs of isolines were identified as being suitable for physiological studies, because the two lines in each pair showed similar growth, N assimilation, and flowering traits under nonsymbiotic conditions. Tests of those lines under N2-dependent conditions with isogenic Hup+ and negligible Hup (Hup−) mutants of R. leguminosarum 128C53 showed that, in symbioses with Hup+ rhizobia, two out of three Hup++ pea lines decreased N2 fixation relative to Hup+ peas. In one of those cases, however, the Hup++ plant line also decreased fixation by Hup− rhizobia. When results were averaged across all rhizobia tested, Hup+ pea isolines had 8.2% higher dry weight (P ≤ 0.05) and fixed 12.6% more N2 (P ≤ 0.05) than Hup++ isolines. Pea lines described here may help identify host plant factors that influence rhizobial Hup activity and should assist in clarifying how Hup systems

  20. Das Lektin aus der Erbse Pisum sativum : Bindungsstudien, Monomer-Dimer-Gleichgewicht und Rückfaltung aus Fragmenten

    NASA Astrophysics Data System (ADS)

    Küster, Frank

    2002-11-01

    Das Lektin aus Pisum sativum, der Gartenerbse, ist Teil der Familie der Leguminosenlektine. Diese Proteine haben untereinander eine hohe Sequenzhomologie, und die Struktur ihrer Monomere, ein all-ß-Motiv, ist hoch konserviert. Dagegen gibt es innerhalb der Familie eine große Vielfalt an unterschiedlichen Quartärstrukturen, die Gegenstand kristallographischer und theoretischer Arbeiten waren. Das Erbsenlektin ist ein dimeres Leguminosenlektin mit einer Besonderheit in seiner Struktur: Nach der Faltung in der Zelle wird aus einem Loop eine kurze Aminosäuresequenz herausgeschnitten, so dass sich in jeder Untereinheit zwei unabhängige Polypeptidketten befinden. Beide Ketten sind aber stark miteinander verschränkt und bilden eine gemeinsame strukturelle Domäne. Wie alle Lektine bindet Erbsenlektin komplexe Oligosaccharide, doch sind seine physiologische Rolle und der natürliche Ligand unbekannt. In dieser Arbeit wurden Versuche zur Entwicklung eines Funktionstests für Erbsenlektin durchgeführt und seine Faltung, Stabilität und Monomer-Dimer-Gleichgewicht charakterisiert. Um die spezifische Rolle der Prozessierung für Stabilität und Faltung zu untersuchen, wurde ein unprozessiertes Konstrukt in E. coli exprimiert und mit der prozessierten Form verglichen. Beide Proteine zeigen die gleiche kinetische Stabilität gegenüber chemischer Denaturierung. Sie denaturieren extrem langsam, weil nur die isolierten Untereinheiten entfalten können und das Monomer-Dimer-Gleichgewicht bei mittleren Konzentrationen an Denaturierungsmittel auf der Seite der Dimere liegt. Durch die extrem langsame Entfaltung zeigen beide Proteine eine apparente Hysterese im Gleichgewichtsübergang, und es ist nicht möglich, die thermodynamische Stabilität zu bestimmen. Die Stabilität und die Geschwindigkeit der Assoziation und Dissoziation in die prozessierten bzw. nichtprozessierten Untereinheiten sind für beide Proteine gleich. Darüber hinaus konnte gezeigt werden, dass auch unter

  1. Induction of host defences by Rhizobium during ineffective nodulation of pea (Pisum sativum L.) carrying symbiotically defective mutations sym40 (PsEFD), sym33 (PsIPD3/PsCYCLOPS) and sym42.

    PubMed

    Ivanova, Kira A; Tsyganova, Anna V; Brewin, Nicholas J; Tikhonovich, Igor A; Tsyganov, Viktor E

    2015-11-01

    Rhizobia are able to establish a beneficial interaction with legumes by forming a new organ, called the symbiotic root nodule, which is a unique ecological niche for rhizobial nitrogen fixation. Rhizobial infection has many similarities with pathogenic infection and induction of defence responses accompanies both interactions, but defence responses are induced to a lesser extent during rhizobial infection. However, strong defence responses may result from incompatible interactions between legumes and rhizobia due to a mutation in either macro- or microsymbiont. The aim of this research was to analyse different plant defence reactions in response to Rhizobium infection for several pea (Pisum sativum) mutants that result in ineffective symbiosis. Pea mutants were examined by histochemical and immunocytochemical analyses, light, fluorescence and transmission electron microscopy and quantitative real-time PCR gene expression analysis. It was observed that mutations in pea symbiotic genes sym33 (PsIPD3/PsCYCLOPS encoding a transcriptional factor) and sym40 (PsEFD encoding a putative negative regulator of the cytokinin response) led to suberin depositions in ineffective nodules, and in the sym42 there were callose depositions in infection thread (IT) and host cell walls. The increase in deposition of unesterified pectin in IT walls was observed for mutants in the sym33 and sym42; for mutant in the sym42, unesterified pectin was also found around degrading bacteroids. In mutants in the genes sym33 and sym40, an increase in the expression level of a gene encoding peroxidase was observed. In the genes sym40 and sym42, an increase in the expression levels of genes encoding a marker of hypersensitive reaction and PR10 protein was demonstrated. Thus, a range of plant defence responses like suberisation, callose and unesterified pectin deposition as well as activation of defence genes can be triggered by different pea single mutations that cause perception of an otherwise

  2. Biochemical Evidence for the Role of the Waxy Protein from Pea (Pisum sativum L.) as a Granule-Bound Starch Synthase.

    PubMed

    Sivak, M. N.; Wagner, M.; Preiss, J.

    1993-12-01

    Proteins were solubilized from starch extracted from developing pea (Pisum sativum L.) embryos and chromatography of these proteins on a Mono-Q column separated two peaks of starch synthase activity. The major activity peak comprised more than 80% of the total activity. This fraction contained only the Waxy protein, as shown by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate followed by staining for proteins or by immunoblot. A 77-kD polypeptide associated with the starch granules and presumed by others to be a starch synthase could not be detected in any of the active fractions. The native molecular weight of the solubilized starch synthase was 59,600 [plus or minus] 1700 as determined by sucrose density gradient. It is concluded that in pea seeds the Waxy protein and the starch synthase bound to the granule are the same protein.

  3. [Activity of agglutinin inhibitor of the kujavian pea (Pisum sativum L.) in mothers' blood and umbilical cord blood considering the course of pregnancy and delivery].

    PubMed

    Lange-Konior, K

    1999-01-01

    The aim of the paper was to evaluate the activity of inhibitor of the phytoagglutinin Pisum sativum (IfPs) in sera of mothers' and umbilical blood of their newborns in confrontation with the course of pregnancy and delivery. The investigations involved 152 tests of sera collected from women delivering at Department of Obstetrics and Perinatology in the Institute of Gynecology and Obstetrics PMU in Szczecin in the years 1992-1993, as well as 156 samples of sera stemming from their newborn infants and were taken from the umbilical cord vessels. The method of investigations being used in the paper was the reaction of inhibiting the phytohemagglutination, wherein the inhibiting action of sera in bearing women and of sera in umbilical blood exerted on agglutinating one was assessed in relation to human erythrocytes of the group 0 with Pisum sativum lectin properties. The accepted titer of inhibitor of the agglutinin Pisum sativum (IfPs) was expressed as the highest dilution of serum, at which complete inhibition of phytohemagglutination was still preserved. The performed investigations have disclosed statistically significant differences between the activity of IfPs occurring in sera of the mothers and the inhibiting factor in umbilical blood sera of the newborns (Tab. 1). No effect of the duration of pregnancy and the course of pregnancy on the IfPs activity in sera of mothers was disclosed. The absence of inhibitor of Pisum sativum lectin in umbilical blood sera was essentially frequently recorded in premature termination of pregnancy between 31-37 weeks of its duration as well as in sera of newborns born by cesarean section and newborns with birth mass being equal or lower than 2500 g in comparison to sera of full term newborns born by forces of nature (Tab. 2, 3, 5). The birth status of newborns according to Apgar scale did not have any influence of IfPs activity in their sera, however, IfPs activity in sera of umbilical blood was statistically significantly more

  4. Polysaccharide fraction from higher plants which strongly interacts with the cytosolic phosphorylase isozyme. I. Isolation and characterization. [Spinacia oleracea L. ; Pisum sativum L

    SciTech Connect

    Yang, Yi; Steup, M. )

    1990-11-01

    From leaves of Spinacia oleracea L. or from Pisum sativum L. and from cotyledons of germinating pea seeds a high molecular weight polysaccharide fraction was isolated. The apparent size of the fraction, as determined by gel filtration, was similar to that of dextran blue. Following acid hydrolysis the monomer content of the polysaccharide preparation was studied using high pressure liquid and thin layer chromatography. Glucose, galactose, arabinose, and ribose were the main monosaccharide compounds. The native polysaccharide preparation interacted strongly with the cytosolic isozyme of phosphorylase (EC 2.4.1.1). Interaction with the plastidic phosphorylase isozyme(s) was by far weaker. Interaction with the cytosolic isozyme was demonstrated by affinity electrophoresis, kinetic measurements, and by {sup 14}C-labeling experiments in which the glucosyl transfer from ({sup 14}C)glucose 1-phosphate to the polysaccharide preparation was monitored.

  5. Combinatorial variation in coding and promoter sequences of genes at the Tri locus in Pisum sativum accounts for variation in trypsin inhibitor activity in seeds.

    PubMed

    Page, D; Aubert, G; Duc, G; Welham, T; Domoney, C

    2002-05-01

    Cultivars of Pisum sativum that differ with respect to the quantitative expression of trypsin/chymotrypsin inhibitor proteins in seeds have been examined in terms of the structure of the corresponding genes. The patterns of divergence in the promoter and coding sequences are described, and the divergence among these exploited for the development of facile DNA-based assays to distinguish genotypes. Quantitative effects on gene expression may be attributed to the overall gene complement and to particular promoter/coding sequence combinations, as well as to the existence of distinct active-site variants that ultimately influence protein activity. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s00438-002-0667-4.

  6. Influence of s-Triazines on Some Enzymes of Carbohydrates and Nitrogen Metabolism in Leaves of Pea (Pisum sativum L.) and Sweet Corn (Zea mays L.)

    PubMed Central

    Wu, M. T.; Singh, B.; Salunkhe, D. K.

    1971-01-01

    Foliar applications of 2 milligrams per liter of 2-chloro-4,6-bis (ethylamino)-s-triazine, 2-methylmercapto-4-ethylamino-6-isobutylamino-s-triazine, and 2-methoxy-4-isopropylamino-6-butylamino-s-triazine caused increases in the activities of starch phosphorylase, pyruvate kinase, cytochrome oxidase, and glutamate dehydrogenase 5, 10, and 15 days after treatment in the leaves of 3-week-old seedlings of pea (Pisum sativum L.) and sweet corn (Zea mays L.). The results indicate that sublethal concentrations of s-triazine compounds affect the physiological and biochemical events in plants which favor more utilization of carbohydrates for nitrate reduction and synthesis of amino acids and proteins. PMID:16657830

  7. Stomatal closure induced by phytosphingosine-1-phosphate and sphingosine-1-phosphate depends on nitric oxide and pH of guard cells in Pisum sativum.

    PubMed

    Puli, Mallikarjuna Rao; Rajsheel, Pidakala; Aswani, Vetcha; Agurla, Srinivas; Kuchitsu, Kazuyuki; Raghavendra, Agepati S

    2016-10-01

    Phyto-S1P and S1P induced stomatal closure in epidermis of pea ( Pisum sativum ) by raising the levels of NO and pH in guard cells. Phosphosphingolipids, such as phytosphingosine-1-phosphate (phyto-S1P) and sphingosine-1-phosphate (S1P), are important signaling components during drought stress. The biosynthesis of phyto-S1P or S1P is mediated by sphingosine kinases (SPHKs). Although phyto-S1P and S1P are known to be signaling components in higher plants, their ability to induce stomatal closure has been ambiguous. We evaluated in detail the effects of phyto-S1P, S1P and SPHK inhibitors on signaling events leading to stomatal closure in the epidermis of Pisum sativum. Phyto-S1P or S1P induced stomatal closure, along with a marked rise in nitric oxide (NO) and cytoplasmic pH of guard cells, as in case of ABA. Two SPHK inhibitors, DL-threo dihydrosphingosine and N',N'-dimethylsphingosine, restricted ABA-induced stomatal closure and prevented the increase of NO or pH by ABA. Modulators of NO or pH impaired both stomatal closure and increase in NO or pH by phyto-S1P/S1P. The stomatal closure by phyto-S1P/S1P was mediated by phospholipase D and phosphatidic acid (PA). When present, PA elevated the levels of pH, but not NO of guard cells. Our results demonstrate that stomatal closure induced by phyto-S1P and S1P depends on rise in pH as well as NO of guard cells. A scheme of signaling events initiated by phyto-S1P/S1P, and converging to cause stomatal closure, is proposed.

  8. Pea (Pisum sativum L.)

    USDA-ARS?s Scientific Manuscript database

    Pea belongs to the Leguminosae plant family, the third largest flowering plant family with 800 genera and over 18,000 species. Tribe Fabeae is considered one of the youngest groups in the legumes and Bayesian molecular clock and ancestral range analysis suggest a crown age of 23 – 16 Mya, in the mi...

  9. Peas (Pisum sativum L.).

    PubMed

    Grant, Jan; Cooper, Pauline

    2006-01-01

    In this chapter we describe a robust method for transformation of peas that has been successfully used in our laboratory since 1992. Using immature pea seed collected from field- or greenhouse-grown plants, we have produced transgenic lines for over 30 genotypes including named pea cultivars and advanced breeding lines. This method uses immature cotyledons as the explant, and the transformation efficiency is in the range 0.2 to 13.5% of cotyledons producing at least one independently transformed line. Agrobacterium tumefaciens strains AGL1 and KYRT1 are the most successful in our procedure, and kanamycin, phosphinothricin, and hygromycin are reliable selectable markers. Potentially useful genes have been introduced for pest and disease resistance, altering quality traits, and investigating metabolic pathways and are being studied in transgenic pea lines.

  10. COCHLEATA controls leaf size and secondary inflorescence architecture via negative regulation of UNIFOLIATA (LEAFY ortholog) gene in garden pea Pisum sativum.

    PubMed

    Sharma, Vishakha; Chaudhary, Swati; Kumar, Arvind; Kumar, Sushil

    2012-12-01

    UNIFOLIATA [(UNI) or UNIFOLIATA-TENDRILLED ACACIA (UNI-TAC)] expression is known to be negatively regulated by COCHLEATA (COCH) in the differentiating stipules and flowers of Pisum sativum. In this study, additional roles of UNI and COCH in P. sativum were investigated. Comparative phenotyping revealed pleiotropic differences between COCH (UNI-TAC and uni-tac) and coch (UNI-TAC and uni-tac) genotypes of common genetic background. Secondary inflorescences were bracteole-less and bracteolated in COCH and coch genotypes, respectively. In comparison to the leaves and corresponding sub-organs and tissues produced on COCH plants, coch plants produced leaves of 1.5-fold higher biomass, 1.5-fold broader petioles and leaflets that were 1.8-fold larger in span and 1.2-fold dorso-ventrally thicker. coch leaflets possessed epidermal cells 1.3-fold larger in number and size, 1.4-fold larger spongy parenchyma cells and primary vascular bundles with 1.2-fold larger diameter. The transcript levels of UNI were at least 2-fold higher in coch leaves and secondary inflorescences than the corresponding COCH organs. It was concluded that COCH negatively regulated UNI in the differentiating leaves and secondary inflorescences and thereby controlled their sizes and/or structures. It was also surmised that COCH and UNI (LFY homolog) occur together widely in stipulate flowering plants.

  11. A novel lipid transfer protein from the pea Pisum sativum: isolation, recombinant expression, solution structure, antifungal activity, lipid binding, and allergenic properties.

    PubMed

    Bogdanov, Ivan V; Shenkarev, Zakhar O; Finkina, Ekaterina I; Melnikova, Daria N; Rumynskiy, Eugene I; Arseniev, Alexander S; Ovchinnikova, Tatiana V

    2016-04-30

    Plant lipid transfer proteins (LTPs) assemble a family of small (7-9 kDa) ubiquitous cationic proteins with an ability to bind and transport lipids as well as participate in various physiological processes including defense against phytopathogens. They also form one of the most clinically relevant classes of plant allergens. Nothing is known to date about correlation between lipid-binding and IgE-binding properties of LTPs. The garden pea Pisum sativum is widely consumed crop and important allergenic specie of the legume family. This work is aimed at isolation of a novel LTP from pea seeds and characterization of its structural, functional, and allergenic properties. Three novel lipid transfer proteins, designated as Ps-LTP1-3, were found in the garden pea Pisum sativum, their cDNA sequences were determined, and mRNA expression levels of all the three proteins were measured at different pea organs. Ps-LTP1 was isolated for the first time from the pea seeds, and its complete amino acid sequence was determined. The protein exhibits antifungal activity and is a membrane-active compound that causes a leakage from artificial liposomes. The protein binds various lipids including bioactive jasmonic acid. Spatial structure of the recombinant uniformly (13)C,(15)N-labelled Ps-LTP1 was solved by heteronuclear NMR spectroscopy. In solution the unliganded protein represents the mixture of two conformers (relative populations ~ 85:15) which are interconnected by exchange process with characteristic time ~ 100 ms. Hydrophobic residues of major conformer form a relatively large internal tunnel-like lipid-binding cavity (van der Waals volume comes up to ~1000 Å(3)). The minor conformer probably corresponds to the protein with the partially collapsed internal cavity. For the first time conformational heterogeneity in solution was shown for an unliganded plant lipid transfer protein. Heat denaturation profile and simulated gastrointestinal digestion assay showed that Ps

  12. ( sup 14 C)-Sucrose uptake by guard cell protoplasts of pisum sativum, argenteum mutant

    SciTech Connect

    Rohrig, K.; Raschke, K. )

    1991-05-01

    Guard cells rely on import for their supply with reduced carbon. The authors tested by silicone oil centrifugation the ability of guard cell protoplasts to accumulated ({sup 14}C)-sucrose. Uptake rates were corrected after measurement of {sup 14}C-sorbitol and {sup 3}H{sub 2}O spaces. Sucrose uptake followed biphasic kinetics, with a high-affinity component below 1 mM external sucrose (apparent K{sub m} 0.8 mM at 25C) and a low-affinity nonsaturable component above. Uptake depended on pH (optimum at pH 5.0). Variations in the concentrations of external KCl, CCCP, and valinomycin indicated that about one-half of the sucrose uptake rate could be related to an electrochemical gradient across the plasmalemma. Total uptake rates measured at 5 mM external sucrose seem to be sufficient to replenish emptied plastids with starch within a few hours.

  13. Pretreatment of Cr(VI)-amended soil with chromate-reducing rhizobacteria decreases plant toxicity and increases the yield of Pisum sativum.

    PubMed

    Soni, Sumit K; Singh, Rakshapal; Singh, Mangal; Awasthi, Ashutosh; Wasnik, Kundan; Kalra, Alok

    2014-05-01

    Pot culture experiments were performed under controlled greenhouse conditions to investigate whether four Cr(VI)-reducing bacterial strains (SUCR44, SUCR140, SUCR186, and SUCR188) were able to decrease Cr toxicity to Pisum sativum plants in artificially Cr(VI)-contaminated soil. The effect of pretreatment of soil with chromate-reducing bacteria on plant growth, chromate uptake, bioaccumulation, nodulation, and population of Rhizobium was found to be directly influenced by the time interval between bacterial treatment and seed sowing. Pretreatment of soil with SUCR140 (Microbacterium sp.) 15 days before sowing (T+15) showed a maximum increase in growth and biomass in terms of root length (93 %), plant height (94 %), dry root biomass (99 %), and dry shoot biomass (99 %). Coinoculation of Rhizobium with SUCR140 further improved the aforementioned parameter. Compared with the control, coinoculation of SUCR140+R showed a 117, 116, 136, and 128 % increase, respectively, in root length, plant height, dry root biomass, and dry shoot biomass. The bioavailability of Cr(VI) decreased significantly in soil (61 %) and in uptake (36 %) in SUCR140-treated plants; the effects of Rhizobium, however, either alone or in the presence of SUCR140, were not significant. The populations of Rhizobium (126 %) in soil and nodulation (146 %) in P. sativum improved in the presence of SUCR140 resulting in greater nitrogen (54 %) concentration in the plants. This study shows the usefulness of efficient Cr(VI)-reducing bacterial strain SUCR140 in improving yields probably through decreased Cr toxicity and improved symbiotic relationship of the plants with Rhizobium. Further decrease in the translocation of Cr(VI) through improved nodulation by Rhizobium in the presence of efficient Cr-reducing bacterial strains could also decrease the accumulation of Cr in shoots.

  14. Evidence for Phytochrome Regulation of Gibberellin A20 3β-Hydroxylation in Shoots of Dwarf (lele) Pisum sativum L. 1

    PubMed Central

    Campell, Bruce R.; Bonner, Bruce A.

    1986-01-01

    The effect of light on the dwarfing allele, le, in Pisum sativum L. was tested as the growth response to gibberellins prior to or beyond the presumed block in the gibberellin biosynthetic pathway. The response to the substrate (GA20), the product (GA1), and a nonendogenous early precursor (steviol) was compared in plants bearing the normal Le and the deficient lele genotypes in plants made low in gibberellin content genetically (nana lines) or by paclobutrazol treatment to tall (cv Alaska) and dwarf (cv Progress) peas. Both genotypes responded to GA1 under red irradiation and in darkness. The lele plants grew in response to GA20 and steviol in darkness but showed a much smaller response when red irradiated. The Le plants responded to GA20 and steviol in both light and darkness. The red effects on lele plants were largely reversible by far-red irradiation. It is concluded that the deficiency in 3β-hydroxylation of GA20 to GA1 in genotype lele is due to a Pfr-induced blockage in the expression of that activity. PMID:16665165

  15. Reactive oxygen species from type-I photosensitized reactions contribute to the light-induced wilting of dark-grown pea (Pisum sativum) epicotyls.

    PubMed

    Hideg, Eva; Vitányi, Beáta; Kósa, Annamária; Solymosi, Katalin; Bóka, Károly; Won, Sungae; Inoue, Yumi; Ridge, Robert W; Böddi, Béla

    2010-04-01

    Type-II, singlet oxygen-mediated photosensitized damage has already been shown to occur in epicotyls of dark-germinated pea (Pisum sativum L.) seedlings upon illumination, resulting in fast turgor loss and wilting. In this study we show evidence that the palette of reactive oxygen species (ROS) is more complex. Hydrogen peroxide, superoxide and hydroxyl radicals are also formed, suggesting the occurrence of type-I reactions as well. Moreover, hydrogen peroxide injection into the epicotyls in the dark was able to provoke wilting directly. Formation of hydroxyl radicals could also be triggered by the addition of hydrogen peroxide in the dark, preferentially in the mid-sections where wilting occurs, showing that potential mediators of a Fenton reaction are present in the epicotyls, but unevenly distributed. Localization of light-inducible ROS formation fully (hydrogen peroxide) or partially (superoxide radicals) overlaps with the distribution of monomer protochlorophyllide complexes, showing that these pigment forms are capable of provoking both type-I and type-II reactions.

  16. Biogenic synthesis of Fe3O4 magnetic nanoparticles using Pisum sativum peels extract and its effect on magnetic and Methyl orange dye degradation studies

    NASA Astrophysics Data System (ADS)

    Prasad, Cheera; Yuvaraja, Gutha; Venkateswarlu, Ponneri

    2017-02-01

    We have been developed facile and ecofriendly method for the synthesis of Fe3O4 magnetic nanoparticles (MNPs) using an aqueous extract of Pisum sativum peels (PS) is used as reducing and capping agent. The as synthesized PS-Fe3O4 MNPs are characterized by diverse techniques such as FTIR, powder XRD, TEM, BET and Raman spectroscopy measurements. The results show that the obtained Fe3O4 nanoparticles exhibits high specific surface area (∼17.6 m2/g) and agglomerated spherical in shape with the size range of 20-30 nm. The magnetic properties of PS-Fe3O4 MNPs sample clearly exhibits ferromagnetic nature with a saturation magnetization of 64.2 emu/g. Further, the catalytic properties of PS-Fe3O4 MNPs for degradation of Methyl orange (MO) dye in aqueous solution have been investigated by UV-visible spectroscopy. The results show that PS-Fe3O4 MNPs is an efficient catalyst for degradation of Methyl orange dye than previously reported ones.

  17. Comparative investigation of concentrations of major and trace elements in organic and conventional Danish agricultural crops. 1. Onions (Allium cepa Hysam) and peas (Pisum sativum ping pong).

    PubMed

    Gundersen, V; Bechmann, I E; Behrens, A; Stürup, S

    2000-12-01

    210 samples of onions (Allium cepa Hysam) from 11 conventionally and 10 organically cultivated sites and 190 samples of peas (Pisum sativum Ping Pong) from 10 conventionally and 9 organically cultivated sites in Denmark were collected and analyzed for 63 and 55 major and trace elements, respectively, by high-resolution inductively coupled plasma mass spectrometry. Sampling, sample preparation, and analysis of the samples were performed under carefully controlled contamination-free conditions. Comparative statistical tests of the element concentration mean values for each site show significantly (p < 0.05) different levels of Ca, Mg, B, Bi, Dy, Eu, Gd, Lu, Rb, Sb, Se, Sr, Ti, U, and Y between the organically and conventionally grown onions and significantly (p < 0.05) different levels of P, Gd, and Ti between the organically and conventionally grown peas. Principal component analysis (PCA) applied to the 63 elements measured in the individual onion samples from the 21 sites split up the sites into two groups according to the cultivation method when the scores of the first and third principal components were plotted against each other. Correspondingly, for peas, a PCA applied to the 55 elements measured as mean values for each site split up the 19 sites into two groups according to the cultivation method when the scores of the third and fourth principal component were plotted against each other. The methodology may be used as authenticity control for organic cultivation after further method development.

  18. The major nucleoside triphosphatase in pea (Pisum sativum L.) nuclei and in rat liver nuclei share common epitopes also present in nuclear lamins

    NASA Technical Reports Server (NTRS)

    Tong, C. G.; Dauwalder, M.; Clawson, G. A.; Hatem, C. L.; Roux, S. J.

    1993-01-01

    The major nucleoside triphosphatase (NTPase) activities in mammalian and pea (Pisum sativum L.) nuclei are associated with enzymes that are very similar both biochemically and immunochemically. The major NTPase from rat liver nuclei appears to be a 46-kD enzyme that represents the N-terminal portion of lamins A and C, two lamina proteins that apparently arise from the same gene by alternate splicing. Monoclonal antibody (MAb) G2, raised to human lamin C, both immunoprecipitates the major (47 kD) NTPase in pea nuclei and recognizes it in western blot analyses. A polyclonal antibody preparation raised to the 47-kD pea NTPase (pc480) reacts with the same lamin bands that are recognized by MAb G2 in mammalian nuclei. The pc480 antibodies also bind to the same lamin-like bands in pea nuclear envelope-matrix preparations that are recognized by G2 and three other MAbs known to bind to mammalian lamins. In immunofluorescence assays, pc480 and anti-lamin antibodies stain both cytoplasmic and nuclear antigens in plant cells, with slightly enhanced staining along the periphery of the nuclei. These results indicate that the pea and rat liver NTPases are structurally similar and that, in pea nuclei as in rat liver nuclei, the major NTPase is probably derived from a lamin precursor by proteolysis.

  19. Increasing the rate of drying reduces metabolic imbalance, lipid peroxidation and critical water content in radicles of garden pea (Pisum sativum L.).

    PubMed

    Ntuli, Tobias M; Pammenter, Norman W; Berjak, Patricia

    2013-01-01

    Orthodox seeds become desiccation-sensitive as they undergo germination. As a result, germinating seeds serve as a model to study desiccation sensitivity in plant tissues. The effects of the rate of drying on the viability, respiratory metabolism and free radical processes were thus studied during dehydration and wet storage of radicles of Pisum sativum. For both drying regimes desiccation could be described by exponential and inverse modified functions. Viability, as assessed by germination capacity and tetrazolium staining, remained at 100% during rapid (< 24 h) desiccation. However, it declined sharply at c. 0.26 g g¹ dm following slow (c. 5 days) drying. Increasing the rate of dehydration thus lowered the critical water content for survival. Rapid desiccation was also associated with higher activities and levels of malate dehydrogenase and the oxidized form of nicotinamide adenine dinucleotide. It was also accompanied by lower hydroperoxide levels and membrane damage. In addition, the activitiy of glutathione reductase was greater during rapid drying. Ageing may have contributed to increased damage during slow dehydration, since viability declined even in wet storage after two weeks. The results presented are consistent with rapid desiccation reducing the accumulation of damage resulting from desiccation-induced aqueous-based deleterious reactions. In addition, they show that radicles are a useful model to study desiccation sensitivity in plant tissues.

  20. Characterization of a rapid, blue light-mediated change in detectable phosphorylation of a plasma membrane protein from etiolated pea (Pisum sativum L. ) seedlings

    SciTech Connect

    Short, T.W.; Briggs, W.R. )

    1990-01-01

    When crude microsomal membranes from apical stem segments of etiolated Pisum sativum L. cv Alaska are mixed in vitro with {gamma}-({sup 32}P)ATP, a phosphorylated band of apparent molecular mass 120 kilodaltons can be detected on autoradiographs of sodium dodecyl sulfate electrophoresis gels. If the stem sections are exposed to blue light immediately prior to membrane isolation, this band is not evident. Comparisons of the kinetics, tissue distribution, and dark recovery of the phosphorylation response with those published for blue light mediated phototropism or rapid growth inhibition indicate that the phosphorylation could be linked to one or both of the reactions described. However, the fluence-response relationships for the change in detectable phosphorylation match quite closely those reported for phototropism but not those for growth inhibition. Blue light has also been found to regulate the capacity for in vitro phosphorylation of a second protein. It has an apparent molecular mass of 84 kilodaltons and is localized primarily in basal stem sections.

  1. Seasonal Patterns of 13C Partitioning Between Shoots and Nodulated Roots of N2‐ or Nitrate‐fed Pisum sativum L.

    PubMed Central

    VOISIN, A. S.; SALON, C.; JEUDY, C.; WAREMBOURG, F. R.

    2003-01-01

    The effect of nitrogen source (N2 or nitrate) on carbon assimilation by photosynthesis and on carbon partitioning between shoots and roots was investigated in pea (Pisum sativum L. ‘Baccara’) plants at different growth stages using 13C labelling. Plants were grown in the greenhouse on different occasions in 1999 and 2000. Atmospheric [CO2] and growth conditions were varied to alter the rate of photosynthesis. Carbon allocation to nodulated roots was unaffected by N source. At the beginning of the vegetative period, nodulated roots had priority for assimilates over shoots; this priority decreased during later stages and became identical to that of the shoot during seed filling. Carbon allocation to nodulated roots was always limited by competition with shoots, and could be predicted for each phenological stage: during vegetative and flowering stages a single, negative exponential relationship was established between sink intensity (percentage of C allocated to the nodulated root per unit biomass) and net photosynthesis. At seed filling, the amount of carbon allocated to the nodulated root was directly related to net photosynthesis. Respiration of nodulated roots accounted for more than 60 % of carbon allocated to them during growth. Only at flowering was respiration affected by N supply: it was significantly higher for strictly N2‐fixing plants (83 %) than for plants fed with nitrate (71 %). At the vegetative stage, the increase in carbon in nodulated root biomass was probably limited by respiration losses. PMID:12646498

  2. A Comparison of the Effects of Chilling on Leaf Gas Exchange in Pea (Pisum sativum L.) and Cucumber (Cucumis sativus L.) 1

    PubMed Central

    Peeler, Thomas C.; Naylor, Aubrey W.

    1988-01-01

    The effects of chilling on the photosynthesis of a chilling-resistant species, pea (Pisum sativum L. cv Alaska) and a chilling-sensitive species, cucumber (Cucumis sativus L. cv Ashley) were compared in order to determine the differences in the photosynthetic chilling sensitivity of these two species. For these experiments, plants were chilled (5°C) for different lengths of time in the dark or light. Following a 1 hour recovery period at 25°C, photosynthetic activity was measured by gas exchange (CO2 uptake and H2O release), quantum yield, and induced chlorophyll fluorescence. The results show that pea photosynthesis was largely unaffected by two consecutive nights of chilling in the dark, or by chilling during a complete light and dark cycle (15 hours/9 hours). Cucumber gas exchange was reduced by one night of chilling, but its quantum yield and variable fluorescence were unaffected by dark chilling. However, chilling cucumber in the light led to reduced CO2 fixation, increased internal leaf CO2 concentration, decreased quantum yield, and loss of variable fluorescence. These results indicate that chilling temperatures in conjunction with light damaged the light reactions of photosynthesis, while chilling in the dark did not. PMID:16665856

  3. The active site architecture of Pisum sativum β-carbonic anhydrase is a mirror image of that of α-carbonic anhydrases

    PubMed Central

    Kimber, Matthew S.; Pai, Emil F.

    2000-01-01

    We have determined the structure of the β–carbonic anhydrase from the dicotyledonous plant Pisum sativum at 1.93 Å resolution, using a combination of multiple anomalous scattering off the active site zinc ion and non-crystallographic symmetry averaging. The mol– ecule assembles as an octamer with a novel dimer of dimers of dimers arrangement. Two distinct patterns of conservation of active site residues are observed, implying two potentially mechanistically distinct classes of β–carbonic anhydrases. The active site is located at the interface between two monomers, with Cys160, His220 and Cys223 binding the catalytic zinc ion and residues Asp162 (oriented by Arg164), Gly224, Gln151, Val184, Phe179 and Tyr205 interacting with the substrate analogue, acetic acid. The substrate binding groups have a one to one correspondence with the functional groups in the α–carbonic anhydrase active site, with the corresponding residues being closely superimposable by a mirror plane. Therefore, despite differing folds, α- and β–carbonic anhydrase have converged upon a very similar active site design and are likely to share a common mechanism. PMID:10747009

  4. Discriminant Analysis of Defective and Non-Defective Field Pea (Pisum sativum L.) into Broad Market Grades Based on Digital Image Features.

    PubMed

    McDonald, Linda S; Panozzo, Joseph F; Salisbury, Phillip A; Ford, Rebecca

    2016-01-01

    Field peas (Pisum sativum L.) are generally traded based on seed appearance, which subjectively defines broad market-grades. In this study, we developed an objective Linear Discriminant Analysis (LDA) model to classify market grades of field peas based on seed colour, shape and size traits extracted from digital images. Seeds were imaged in a high-throughput system consisting of a camera and laser positioned over a conveyor belt. Six colour intensity digital images were captured (under 405, 470, 530, 590, 660 and 850nm light) for each seed, and surface height was measured at each pixel by laser. Colour, shape and size traits were compiled across all seed in each sample to determine the median trait values. Defective and non-defective seed samples were used to calibrate and validate the model. Colour components were sufficient to correctly classify all non-defective seed samples into correct market grades. Defective samples required a combination of colour, shape and size traits to achieve 87% and 77% accuracy in market grade classification of calibration and validation sample-sets respectively. Following these results, we used the same colour, shape and size traits to develop an LDA model which correctly classified over 97% of all validation samples as defective or non-defective.

  5. Spatial and Temporal Influences on the Cell-Specific Distribution of Glycine Decarboxylase in Leaves of Wheat (Triticum aestivum L.) and Pea (Pisum sativum L.) 1

    PubMed Central

    Tobin, Alyson K.; Thorpe, Julian R.; Hylton, Christopher M.; Rawsthorne, Stephen

    1989-01-01

    The distribution of glycine decarboxylase (GDC) in leaves of pea (Pisum sativum L.) and wheat (Triticum aestivum L.) has been investigated using immunogold labeling of the P-protein subunit of the GDC complex. Mitochondria in photosynthetic mesophyll cells were densely labeled, whereas those in nonphotosynthetic vascular parenchyma and epidermal cells were only weakly labeled. In pea leaves the density of immunogold labeling on mitochondria in the chloroplast-containing bundle sheath and stomatal guard cells was intermediate between that in mesophyll and epidermal cells. In both species the density of labeling on mitochondria in a cell appeared to reflect the photosynthetic capacity of the cell. This relationship was further examined in wheat where a natural developmental gradient exists along the lamina such that cell maturity increases with distance from the basal meristem. In this case the density of labeling on mesophyll cell mitochondria increased with photosynthetic development and with increasing maturity of the cell. Vascular cell mitochondria, however, became less densely labeled as the cells matured. The results indicate a close, positive correlation between the concentration of GDC in the mitochondria and the photosynthetic status of the host cell. This relationship is maintained effectively under the influence of both spatial (i.e. cellular differentiation across the lamina) and temporal (i.e. cellular development along the lamina) constraints. Images Figure 1 Figure 2 PMID:16667135

  6. Purification and Characterization of S-Adenosyl-l-methionine:6a-Hydroxymaackiain 3-O-Methyltransferase from Pisum sativum1

    PubMed Central

    Preisig, Carol L.; Matthews, David E.; VanEtten, Hans D.

    1989-01-01

    The isoflavonoid phytoalexin pisatin is synthesized by Pisum sativum in response to microbial infection and certain other forms of stress. An enzyme which synthesizes pisatin by methylating the 3-hydroxyl of (+)6a-hydroxymaackiain (HMK) was extracted from CuCl2-stressed pea seedlings. The enzyme was enriched 370-fold by (NH4)2SO4 precipitation, DEAE chromatography, chromatofocusing, and hydrophobic interaction chromatography (HIC), to a specific activity of 8.2 microkatals per gram protein. Enzyme activity profiles from chromatofocusing and HIC columns suggested the presence of two isozymes, of pl 5.2 and 4.9. Nondenaturing gel filtration of the HIC-purified enzyme gave a single peak of activity at the same elution volume as BSA (66 kilodaltons); the active fractions showed two proteins upon SDS-PAGE, of Mr 66,000 and 43,000. The smaller protein was most abundant in chromatographic fractions containing peak enzyme activity throughout purification. In a partially purified preparation, this 43 kilodalton protein was the only one photoaffinity labelled by [3H]S-adenosyl-l-methionine. The purified enzyme preferred the (+) over the (−) stereoisomer of HMK and other pterocarpans; overall, (+)HMK was the best substrate. Km values were 2.3 micromolar for (+)HMK and 35 micromolar for S-adenosyl-l-methionine. The methyltransferase had a pH optimum of 7.9 and no apparent divalent cation requirement. Images Figure 2 Figure 3 Figure 5 Figure 6 PMID:16667069

  7. The Use of Fura-2 Fluorescence to Monitor the Movement of Free Calcium Ions into the Matrix of Plant Mitochondria (Pisum sativum and Helianthus tuberosus).

    PubMed

    Zottini, M.; Zannoni, D.

    1993-06-01

    Purified mitochondria isolated from pea (Pisum sativum L. cv Alaska) stems and Jerusalem artichoke (Helianthus tuberosus L. cv OB1) tubers were loaded with the acetoxymethyl ester of the fluorescent Ca2+ indicator fura-2. This made possible the continuous monitoring of free [Ca2+] in the matrix ([Ca2+]m) without affecting the apparent viability of the mitochondria. Pea stem mitochondria contained an initial [Ca2+]m of approximately 60 to 100 nM, whereas [Ca2+]m was severalfold higher (400-600 nM) in mitochondria of Jerusalem artichoke tubers. At low extramitochondrial Ca2+ concentrations ([greater than or equal to]100 nM), there was an energy-dependent membrane potential increase in [Ca2+]m; the final [Ca2+]m was phosphate-dependent in Jerusalem artichoke but was phosphate-independent in pea stem mitochondria. The data presented indicate that (a) there is no absolute requirement for phosphate in Ca2+ uptake; (b) plant mitochondria can accumulate external free Ca2+ by means of an electrophoretic Ca2+ uniporter with an apparent affinity for Ca2+ (Km approximately 150 nM) that is severalfold lower than that measured by conventional methods (isotopes and Ca2+-sensitive electrodes); and (c) [Ca2+]m is within the regulatory range of mammalian intramitochondrial dehydrogenases.

  8. Nuclear-Cytoplasmic Conflict in Pea (Pisum sativum L.) Is Associated with Nuclear and Plastidic Candidate Genes Encoding Acetyl-CoA Carboxylase Subunits

    PubMed Central

    Bogdanova, Vera S.; Zaytseva, Olga O.; Mglinets, Anatoliy V.; Shatskaya, Natalia V.; Kosterin, Oleg E.; Vasiliev, Gennadiy V.

    2015-01-01

    In crosses of wild and cultivated peas (Pisum sativum L.), nuclear-cytoplasmic incompatibility frequently occurs manifested as decreased pollen fertility, male gametophyte lethality, sporophyte lethality. High-throughput sequencing of plastid genomes of one cultivated and four wild pea accessions differing in cross-compatibility was performed. Candidate genes for involvement in the nuclear-plastid conflict were searched in the reconstructed plastid genomes. In the annotated Medicago truncatula genome, nuclear candidate genes were searched in the portion syntenic to the pea chromosome region known to harbor a locus involved in the conflict. In the plastid genomes, a substantial variability of the accD locus represented by nucleotide substitutions and indels was found to correspond to the pattern of cross-compatibility among the accessions analyzed. Amino acid substitutions in the polypeptides encoded by the alleles of a nuclear locus, designated as Bccp3, with a complementary function to accD, fitted the compatibility pattern. The accD locus in the plastid genome encoding beta subunit of the carboxyltransferase of acetyl-coA carboxylase and the nuclear locus Bccp3 encoding biotin carboxyl carrier protein of the same multi-subunit enzyme were nominated as candidate genes for main contribution to nuclear-cytoplasmic incompatibility in peas. Existence of another nuclear locus involved in the accD-mediated conflict is hypothesized. PMID:25789472

  9. Evaluation of the protection exerted by Pisum sativum Ferredoxin-NADP(H) Reductase against injury induced by hypothermia on Cos-7 cells.

    PubMed

    Pucci Molineris, M; Di Venanzio, G; Mamprin, M E; Mediavilla, M G

    2013-08-01

    Hypothermia is employed as a method to diminish metabolism rates and preserve tissues and cells. However, low temperatures constitute a stress that produces biochemical changes whose extension depends on the duration and degree of cold exposure and is manifested when physiological temperature is restored. For many cellular types, cold induces an oxidative stress that is dependent on the elevation of intracellular iron, damages macromolecules, and is prevented by the addition of iron chelators. Pisum sativum Ferredoxin-NADP(H) Reductase (FNR) has been implicated in protection from injury mediated by intracellular iron increase and successfully used to reduce oxidative damage on bacterial, plant and mammalian systems. In this work, FNR was expressed in Cos-7 cells; then, they were submitted to cold incubation and iron overload to ascertain whether this enzyme was capable of diminishing the harm produced by these challenges. Contrary to expected, FNR was not protective and even exacerbated the damage under certain circumstances. It was also found that the injury induced by hypothermia in Cos-7 cells presented both iron-dependent and iron-independent components of damage when cells were actively dividing but only iron-independent component when cells were in an arrested state. This is in agreement with previous findings which showed that iron-dependent damage is also an energy-dependent process.

  10. Systemic Induction of the Defensin and Phytoalexin Pisatin Pathways in Pea (Pisum sativum) against Aphanomyces euteiches by Acetylated and Nonacetylated Oligogalacturonides.

    PubMed

    Selim, Sameh; Sanssené, Jean; Rossard, Stéphanie; Courtois, Josiane

    2017-06-19

    Oligogalacturonides (OGs) are known for their powerful ability to stimulate the plant immune system but little is known about their mode of action in pea (Pisum sativum). In the present study, we investigated the elicitor activity of two fractions of OGs, with polymerization degrees (DPs) of 2-25, in pea against Aphanomyces euteiches. One fraction was nonacetylated (OGs - Ac) whereas the second one was 30% acetylated (OGs + Ac). OGs were applied by injecting the upper two rachises of the plants at three- and/or four-weeks-old. Five-week-old roots were inoculated with 10⁵ zoospores of A. euteiches. The root infection level was determined at 7, 10 and 14 days after inoculation using the quantitative real-time polymerase chain reaction (qPCR). Results showed significant root infection reductions namely 58, 45 and 48% in the plants treated with 80 µg OGs + Ac and 59, 56 and 65% with 200 µg of OGs - Ac. Gene expression results showed the upregulation of genes involved in the antifungal defensins, lignans and the phytoalexin pisatin pathways and a priming effect in the basal defense, SA and ROS gene markers as a response to OGs. The reduction of the efficient dose in OGs + Ac is suggesting that acetylation is necessary for some specific responses. Our work provides the first evidence for the potential of OGs in the defense induction in pea against Aphanomyces root rot.

  11. Genetic Changes Accompanying the Domestication of Pisum sativum: Is there a Common Genetic Basis to the ‘Domestication Syndrome’ for Legumes?

    PubMed Central

    Weeden, Norman F.

    2007-01-01

    Background and Aims The changes that occur during the domestication of crops such as maize and common bean appear to be controlled by relatively few genes. This study investigates the genetic basis of domestication in pea (Pisum sativum) and compares the genes involved with those determined to be important in common bean domestication. Methods Quantitative trait loci and classical genetic analysis are used to investigate and identify the genes modified at three stages of the domestication process. Five recombinant inbred populations involving crosses between different lines representing different stages are examined. Key Results A minimum of 15 known genes, in addition to a relatively few major quantitative trait loci, are identified as being critical to the domestication process. These genes control traits such as pod dehiscence, seed dormancy, seed size and other seed quality characters, stem height, root mass, and harvest index. Several of the genes have pleiotropic effects that in species possessing a more rudimentary genetic characterization might have been interpreted as clusters of genes. Very little evidence for gene clustering was found in pea. When compared with common bean, pea has used a different set of genes to produce the same or similar phenotypic changes. Conclusions Similar to results for common bean, relatively few genes appear to have been modified during the domestication of pea. However, the genes involved are different, and there does not appear to be a common genetic basis to ‘domestication syndrome’ in the Fabaceae. PMID:17660515

  12. Identification of Phenolic Compounds from Seed Coats of Differently Colored European Varieties of Pea (Pisum sativum L.) and Characterization of Their Antioxidant and In Vitro Anticancer Activities.

    PubMed

    Stanisavljević, Nemanja S; Ilić, Marija D; Matić, Ivana Z; Jovanović, Živko S; Čupić, Tihomir; Dabić, Dragana Č; Natić, Maja M; Tešić, Živoslav Lj

    2016-01-01

    To date little has been done on identification of major phenolic compounds responsible for anticancer and antioxidant properties of pea (Pisum sativum L.) seed coat extracts. In the present study, phenolic profile of the seed coat extracts from 10 differently colored European varieties has been determined using ultrahigh-performance liquid chromatography-linear trap quadrupole orbitrap mass spectrometer technique. Extracts of dark colored varieties with high total phenolic content (up to 46.56 mg GAE/g) exhibited strong antioxidant activities (measured by 2,2-diphenyl-1-picrylhydrazyl or DPPH assay, and ferric ion reducing and ferrous ion chelating capacity assays) which could be attributed to presence of gallic acid, epigallocatechin, naringenin, and apigenin. The aqueous extracts of dark colored varieties exert concentration-dependent cytotoxic effects on all tested malignant cell lines (human colon adenocarcinoma LS174, human breast carcinoma MDA-MB-453, human lung carcinoma A594, and myelogenous leukemia K562). Correlation analysis revealed that intensities of cytotoxic activity of the extracts strongly correlated with contents of epigallocatechin and luteolin. Cell cycle analysis on LS174 cells in the presence of caspase-3 inhibitor points out that extracts may activate other cell death modalities besides caspase-3-dependent apoptosis. The study provides evidence that seed coat extracts of dark colored pea varieties might be used as potential cancer-chemopreventive and complementary agents in cancer therapy.

  13. Quantification of Pea enation mosaic virus 1 and 2 during infection of Pisum sativum by one step real-time RT-PCR.

    PubMed

    Doumayrou, Juliette; Sheber, Melissa; Bonning, Bryony C; Miller, W Allen

    2017-02-01

    Pea enation mosaic virus 1 (PEMV1) and Pea enation mosaic virus 2 (PEMV2) are two viruses in an obligate symbiosis that cause pea enation mosaic disease mainly in plants in the Fabaceae family. This virus system is a valuable model to investigate plant virus replication, movement and vector transmission. Thus, here we describe growth conditions, virus detection methods, and virus accumulation behavior. To measure the accumulation and movement of PEMV1 and PEMV2 in plants during the course of infection, we developed a quantitative real-time one-step reverse transcription PCR procedure using the SYBR-green(®) technology. Viral primers were designed that anneal to conserved but distinct regions in the RNA-dependent RNA polymerase gene of each virus. Moreover, the normalization of viral accumulation was performed to correct for sample-to-sample variation by designing primers to two different Pisum sativum housekeeping genes: actin and β-tubulin. Transcript levels for these housekeeping genes did not change significantly in response to PEMV infection. Conditions were established for maximum PCR efficiency for each gene, and quantification using QuBit(®) technology. Both viruses reached maximum accumulation around 21days post-inoculation of pea plants. These results provide valuable tools and knowledge to allow reproducible studies of this emerging model virus system virus complex. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Role of cyanide-resistant respiration in the response of two pea hybrids to CO/sub 2/ enrichment. [Pisum sativum

    SciTech Connect

    Musgrave, M.E.; Strain, B.R.; Siedow, J.N.

    1986-04-01

    Two cultivars of pea (Pisum sativum L., cvs. Alaska and Progress No. 9) were reciprocally crossed to yield hybrids differing in the presence or absence of the cyanide-resistant (alternative) pathway of respiration. The growth of this material in greenhouses maintained at either 350 or 650 ppm CO/sub 2/ was compared. The objective was to assess the significance of the alternative pathway to whole plant carbon budgets and further to use CO/sub 2/ enrichment as a means of testing the hypothesis that the alternative pathway might be important in oxidizing excess carbohydrates. More structural and storage carbohydrates were available in the cross lacking the pathway than in the reciprocal cross as shown by greater plant height, leaf area, specific leaf weight, branching, total dry matter and seed production. Specific leaf weight increased strongly under CO/sub 2/ enrichment in the hybrid lacking the pathway while it was the same at 350 and 650 ppm in the reciprocal cross. These results suggest that respiration via the alternative pathway may be energetically wasteful in terms of whole plant carbon budgets.

  15. Etioplasts with protochlorophyll and protochlorophyllide forms in the under-soil epicotyl segments of pea (Pisum sativum) seedlings grown under natural light conditions.

    PubMed

    Vitányi, Beáta; Kósa, Annamária; Solymosi, Katalin; Böddi, Béla

    2013-06-01

    To study if etiolation symptoms exist in plants grown under natural illumination conditions, under-soil epicotyl segments of light-grown pea (Pisum sativum) plants were examined and compared to those of hydroponically dark-grown plants. Light-, fluorescence- and electron microscopy, 77 K fluorescence spectroscopy, pigment extraction and pigment content determination methods were used. Etioplasts with prolamellar bodies and/or prothylakoids, protochlorophyll (Pchl) and protochlorophyllide (Pchlide) forms (including the flash-photoactive 655 nm emitting form) were found in the (pro)chlorenchyma of epicotyl segments under 3 cm soil depth; their spectral properties were similar to those of hydroponically grown seedlings. However, differences were found in etioplast sizes and Pchlide:Pchl molar ratios, which indicate differences in the developmental rates of the under-soil and of hydroponically developed cells. Tissue regions closer to the soil surface showed gradual accumulation of chlorophyll, and in parallel, decrease of Pchl and Pchlide. These results proved that etioplasts and Pchlide exist in soil-covered parts of seedlings even if they have a 3-4-cm long photosynthetically active shoot above the soil surface. This underlines that etiolation symptoms do develop under natural growing conditions, so they are not merely artificial, laboratory phenomena. Consequently, dark-grown laboratory plants are good models to study the early stages of etioplast differentiation and the Pchlide-chlorophyllide phototransformation. Copyright © Physiologia Plantarum 2012.

  16. The major nucleoside triphosphatase in pea (Pisum sativum L.) nuclei and in rat liver nuclei share common epitopes also present in nuclear lamins

    NASA Technical Reports Server (NTRS)

    Tong, C. G.; Dauwalder, M.; Clawson, G. A.; Hatem, C. L.; Roux, S. J.

    1993-01-01

    The major nucleoside triphosphatase (NTPase) activities in mammalian and pea (Pisum sativum L.) nuclei are associated with enzymes that are very similar both biochemically and immunochemically. The major NTPase from rat liver nuclei appears to be a 46-kD enzyme that represents the N-terminal portion of lamins A and C, two lamina proteins that apparently arise from the same gene by alternate splicing. Monoclonal antibody (MAb) G2, raised to human lamin C, both immunoprecipitates the major (47 kD) NTPase in pea nuclei and recognizes it in western blot analyses. A polyclonal antibody preparation raised to the 47-kD pea NTPase (pc480) reacts with the same lamin bands that are recognized by MAb G2 in mammalian nuclei. The pc480 antibodies also bind to the same lamin-like bands in pea nuclear envelope-matrix preparations that are recognized by G2 and three other MAbs known to bind to mammalian lamins. In immunofluorescence assays, pc480 and anti-lamin antibodies stain both cytoplasmic and nuclear antigens in plant cells, with slightly enhanced staining along the periphery of the nuclei. These results indicate that the pea and rat liver NTPases are structurally similar and that, in pea nuclei as in rat liver nuclei, the major NTPase is probably derived from a lamin precursor by proteolysis.

  17. In situ expression of two storage protein genes in relation to histo-differentiation at mid-embryogenesis in Medicago truncatula and Pisum sativum seeds.

    PubMed

    Abirached-Darmency, M; Abdel-gawwad, M R; Conejero, G; Verdeil, J L; Thompson, R

    2005-08-01

    The seed consists of several layers of specialized cell-types that divide and differentiate following a highly regulated programme in time and space. A cytological approach was undertaken in order to study the histo-differentiation at mid-embryogenesis in Medicago truncatula as a model legume, and in Pisum sativum using serial sections of embedded immature seed. Little published information is available about seed development in Medicago species. The observations from this study revealed a number of distinctive features of Medicago seed development and differentiation. Transfer cells, involved in nutrient transfer to the embryo, were clearly identified in the thin-walled parenchyma of the innermost integument. Histological Schiff-naphthol enabled carbohydrate accumulation to be followed in the different seed compartments, and revealed the storage protein bodies. Non-radioactive mRNA in situ hybridization, was carried out using mRNA probes from two highly expressed genes encoding the major vicilin and legumin A storage protein types. The timing of mRNA expression was related to that of the corresponding proteins already identified.

  18. Physiological changes in Triticum durum, Zea mays, Pisum sativum and Lens esculenta cultivars, caused by irrigation with water contaminated with microcystins: a laboratory experimental approach.

    PubMed

    Saqrane, Sana; Ouahid, Youness; El Ghazali, Issam; Oudra, Brahim; Bouarab, Lahcen; del Campo, Francisca F

    2009-06-01

    The aim of the present study was to investigate the effect of exposure to a microcystin (MC)-containing extract from a cyanobacteria bloom on growth, development, mineral nutrient accumulation, and photosynthetic activity of Triticum durum, Zea mays, Pisum sativum and Lens esculenta cultivars. The MCs in the extract, identified by HPLC and/or mass spectrometry (MS) were: MC-RR, -LR, -YR, -(H4)YR, -WR, and -FR. Plant growth and development was tested along 30 exposure days. After this period, MC-extract caused a clear reduction in plant growth and productivity, as well as deleterious effects on development and Photosystem II activity, measured by Fv/Fm fluorescence. However, the chlorophyll (a + b) content hardly varied, and the accumulation of Na+, K+, Ca2+, P and N was enhanced. All the effects observed were plant species, MC concentration, and exposure-time dependent. Relative accumulation of each MC variant greatly varied among plant species and plant organ. The data obtained supports the idea that the use of surface water containing MCs for crop irrigation can affect both plant yield and quality, and secondly, that MC accumulation in edible plants might pose a potential risk for human and animal health, if the MC intake exceeded the recommended tolerable limits.

  19. Aqueous pathways dominate permeation of solutes across Pisum sativum seed coats and mediate solute transport via diffusion and bulk flow of water.

    PubMed

    Niemann, Sylvia; Burghardt, Markus; Popp, Christian; Riederer, Markus

    2013-05-01

    The permeability of seed coats to solutes either of biological or anthropogenic origin plays a major role in germination, seedling growth and seed treatment by pesticides. An experimental set-up was designed for investigating the mechanisms of seed coat permeation, which allows steady-state experiments with isolated seed coats of Pisum sativum. Permeances were measured for a set of organic model compounds with different physicochemical properties and sizes. The results show that narrow aqueous pathways dominate the diffusion of solutes across pea seed coats, as indicated by a correlation of permeances with the molecular sizes of the compounds instead of their lipophilicity. Further indicators for an aqueous pathway are small size selectivity and a small effect of temperature on permeation. The application of an osmotic water potential gradient across isolated seed coats leads to an increase in solute transfer, indicating that the aqueous pathways form a water-filled continuum across the seed coat allowing the bulk flow of water. Thus, the uptake of organic solutes across pea testae has two components: (1) by diffusion and (2) by bulk water inflow, which, however, is relevant only during imbibition. © 2012 Blackwell Publishing Ltd.

  20. Molecular cloning of isoflavone reductase from pea (Pisum sativum L.): evidence for a 3R-isoflavanone intermediate in (+)-pisatin biosynthesis.

    PubMed

    Paiva, N L; Sun, Y; Dixon, R A; VanEtten, H D; Hrazdina, G

    1994-08-01

    Isoflavone reductase (IFR) reduces achiral isoflavones to chiral isoflavanones during the biosynthesis of chiral pterocarpan phytoalexins. A cDNA clone for IFR from pea (Pisum sativum) was isolated using the polymerase chain reaction and expressed in Escherichia coli. Analysis of circular dichroism (CD) spectra of the reduction product sophorol obtained using the recombinant enzyme indicated that the isoflavanone possessed the 3R stereochemistry, in contrast to previous reports indicating a 3S-isoflavanone as the product of the pea IFR. Analysis of CD spectra of sophorol produced using enzyme extracts of CuCl2-treated pea seedlings confirmed the 3R stereochemistry. Thus, the stereochemistry of the isoflavanone intermediate in (+)-pisatin biosynthesis in pea is the same as that in (-)-medicarpin biosynthesis in alfalfa, although the final pterocarpans have the opposite stereochemistry. At the amino acid level the pea IFR cDNA was 91.8 and 85.2% identical to the IFRs from alfalfa and chickpea, respectively. IFR appears to be encoded by a single gene in pea. Its transcripts are highly induced in CuCl2-treated seedlings, consistent with the appearance of IFR enzyme activity and pisatin accumulation.

  1. Interspecific variation in SO/sub 2/ flux: leaf surface versus internal flux, and components of leaf conductance. [Pisum sativum L. , Lycopersicon esculentum Mill. Flacca, Geranium carolinianum L. , Diplacus aurantiacus (Curtis) Jeps

    SciTech Connect

    Olszyk, D.M.; Tingey, D.T.

    1985-12-01

    The objective of this study was to clarify the relationships among stomatal, residual, and epidermal conductances in determining the flux of SO/sub 2/ air pollution to leaves. Variations in leaf SO/sub 2/ and H/sub 2/O vapor fluxes were determined using four plant species: Pisum sativum L. (garden pea), Lycopersicon esculentum Mill. flacca (mutant of tomato), Geranium carolinianum L. (wild geranium), and Diplacus aurantiacus (Curtis) Jeps. (a native California shrub). Fluxes were measured using the mass-balance approach during exposure to 4.56 micromoles per cubic meter (0.11 microliters per liter) SO/sub 2/ for 2 hours in a controlled environmental chamber. Flux through adaxial and abaxial leaf surfaces with closed stomata ranged from 1.9 to 9.4 nanomoles per square meter per second for SO/sub 2/, and 0.3 to 1.3 millimoles per square meter per second for H/sub 2/O vapor. Flux of SO/sub 2/ into leaves through stomata ranged from approx.0 to 8.5 (dark) and 3.8 to 16.0 (light) millimoles per square meter per second. Flux of H/sub 2/O vapor from leaves through stomata ranged from approx.0 to 0.6 (dark) to 0.4 to 0.9 (light) millimole per square meter per second. Lycopersicon had internal flux rates for both SO/sub 2/ and H/sub 2/O vapor over twice as high as for the other species. Stomatal conductance based on H/sub 2/O vapor flux averaged from 0.07 to 0.13 mole per square meter per second among the four species. Internal conductance of SO/sub 2/ as calculated from SO/sub 2/ flux was from 0.04 mole per square meter per second lower to 0.06 mole per square meter per second higher than stomatal conductance. For Pisum, Geranium, and Diplacus stomatal conductance was the same or slightly higher than internal conductance, indicating that, in general, SO/sub 2/ flux could be predicted from stomatal conductance for H/sub 2/O vapor.

  2. Comparative response of Pisum sativum nodulated with indigenous soil Rhizobium populations and/or co-inoculated with a Rhizobium leguminosarum strain. I. Acetylene-reducing, dihydrogen- and carbon dioxide-evolving activities.

    PubMed

    Skrdleta, V; Nĕmcová, M; Lisá, L; Novák, K; Kovárová, D

    1991-01-01

    No significant differences in the acetylene-reducing activity and evolution of H2 and CO2 from nodulated roots of Pisum sativum inoculated with soil Rhizobium populations from two soils with different acidities (Ruzynĕ soil 7.6; Lukavec soil 4.9) were observed. Rhizobium population from Lukavec soil formed nodules, exhibiting a higher H2 evolution. Co-inoculation with the Hup+ strain 128C30 (7 x 10(7) cells per seedling) eliminated, to some extent, the effect of soil populations on physiological activity.

  3. Gas exchange and chlorophyll fluorescence of pea (Pisum sativum L.) plants in response to ambient ozone at a rural site in Egypt.

    PubMed

    Ismail, I M; Basahi, J M; Hassan, I A

    2014-11-01

    Egyptian pea cultivars (Pisum sativum L. cultivars Little Marvel, Perfection and Victory) grown in open-top chambers were exposed to either charcoal-filtered (FA) or non-filtered air (NF) for five consecutive years (2009-2013) at a rural site in northern Egypt. Net photosynthetic rates (PN), stomatal conductance (gs), intercellular CO2 (Ci) and chlorophyll fluorescence were measured. Ozone (O3) was found to be the most prevalent pollutant common at the rural site and is suspected to be involved in the alteration of the physiological parameters measured in the present investigation. PN of different cultivars were found to respond similarly; decreases of 23, 29 and 39% were observed in the cultivars Perfection, Little Marvel and Victory, respectively (averaged over the five years) due to ambient O3. The maximum impairment in PN was recorded in the cultivar Victory (46%) in 2013 when the highest O3 levels were recorded (90 nL L(-1)). The average stomatal conductance decreased by 20 and 18% in the cultivars Little Marvel and Perfection, respectively, while the average stomatal conductance increased on average by 27% in the cultivar Victory. A significant correlation was found between PN and Ci, indicating the importance of non-stomatal limitations of photosynthesis, especially in the cultivar Victory. The PN vs. Ci curves were fitted to a non-rectangular hyperbolic model. The actual quantum yield (ΦPSII) and photochemical quenching coefficient (qP) were significantly decreased in the leaves of plants exposed to NF air. Non-photochemical quenching (NPQ) was increased in all cultivars. Exposure to NF air caused reductions in chlorophyll (Chl a) of 19, 16 and 30% in the Little Marvel, Perfection and Victory cultivars, respectively.

  4. Multiple impacts of the plant growth-promoting rhizobacterium Variovorax paradoxus 5C-2 on nutrient and ABA relations of Pisum sativum.

    PubMed

    Jiang, Fan; Chen, Lin; Belimov, Andrey A; Shaposhnikov, Alexander I; Gong, Fan; Meng, Xu; Hartung, Wolfram; Jeschke, Dieter W; Davies, William J; Dodd, Ian C

    2012-11-01

    Resolving the physiological mechanisms by which rhizobacteria enhance plant growth is difficult, since many such bacteria contain multiple plant growth-promoting properties. To understand further how the 1-aminocyclopropane-1-carboxylate (ACC) deaminase (ACCd)-containing rhizobacterium Variovorax paradoxus 5C-2 affects plant growth, the flows and partitioning of mineral nutrients and abscisic acid (ABA) and ABA metabolism were studied in pea (Pisum sativum) plants following rhizosphere bacterial inoculation. Although root architecture was not affected, inoculation increased root and shoot biomass, and stomatal conductance, by 20, 15, and 24%, respectively, and increased N, P, K, Ca, and Mg uptake by 16, 81, 50, 46, and 58%, respectively. P deposition in inoculated plant roots was 4.9 times higher than that in uninoculated controls. Rhizobacterial inoculation increased root to shoot xylem flows and shoot to root phloem flows of K by 1.8- and 2.1-fold, respectively. In control plants, major sinks for K deposition were the roots and upper shoot (43% and 49% of total uptake, respectively), while rhizobacterial inoculation increased K distribution to the lower shoot at the expense of other compartments (xylem, phloem, and upper shoot). Despite being unable to metabolize ABA in vitro, V. paradoxus 5C-2 decreased root ABA concentrations and accumulation by 40-60%. Although inoculation decreased xylem ABA flows, phloem ABA flows increased. Whether bacterial ACCd attenuates root to shoot ABA signalling requires further investigation, since ABA is critical to maintain growth of droughted plants, and ACCd-containing organisms have been advocated as a means of minimizing growth inhibition of plants in drying soil.

  5. Studies on the Control of Ascochyta Blight in Field Peas (Pisum sativum L.) Caused by Ascochyta pinodes in Zhejiang Province, China

    PubMed Central

    Liu, Na; Xu, Shengchun; Yao, Xiefeng; Zhang, Guwen; Mao, Weihua; Hu, Qizan; Feng, Zhijuan; Gong, Yaming

    2016-01-01

    Ascochyta blight, an infection caused by a complex of Ascochyta pinodes, Ascochyta pinodella, Ascochyta pisi, and/or Phoma koolunga, is a destructive disease in many field peas (Pisum sativum L.)-growing regions, and it causes significant losses in grain yield. To understand the composition of fungi associated with this disease in Zhejiang Province, China, a total of 65 single-pycnidiospore fungal isolates were obtained from diseased pea samples collected from 5 locations in this region. These isolates were identified as Ascochyta pinodes by molecular techniques and their morphological and physiological characteristics. The mycelia of ZJ-1 could penetrate pea leaves across the stomas, and formed specific penetration structures and directly pierced leaves. The resistance level of 23 available pea cultivars was tested against their representative isolate A. pinodes ZJ-1 using the excised leaf-assay technique. The ZJ-1 mycelia could penetrate the leaves of all tested cultivars, and they developed typical symptoms, which suggested that all tested cultivars were susceptible to the fungus. Chemical fungicides and biological control agents were screened for management of this disease, and their efficacies were further determined. Most of the tested fungicides (11 out of 14) showed high activity toward ZJ-1 with EC50 < 5 μg/mL. Moreover, fungicides, including tebuconazole, boscalid, iprodione, carbendazim, and fludioxonil, displayed more than 80% disease control efficacy under the recorded conditions. Three biocontrol strains of Bacillus sp. and one of Pantoea agglomerans were isolated from pea-related niches and significantly reduced the severity of disease under greenhouse and field conditions. To our knowledge, this is the first study on ascochyta blight in field peas, and results presented here will be useful for controlling the disease in this area. PMID:27148177

  6. Free-air CO2 enrichment (FACE) reduces the inhibitory effect of soil nitrate on N2 fixation of Pisum sativum

    PubMed Central

    Butterly, Clayton R.; Armstrong, Roger; Chen, Deli; Tang, Caixian

    2016-01-01

    Background and Aims Additional carbohydrate supply resulting from enhanced photosynthesis under predicted future elevated CO2 is likely to increase symbiotic nitrogen (N) fixation in legumes. This study examined the interactive effects of atmospheric CO2 and nitrate (NO3–) concentration on the growth, nodulation and N fixation of field pea (Pisum sativum) in a semi-arid cropping system. Methods Field pea was grown for 15 weeks in a Vertosol containing 5, 25, 50 or 90 mg NO3–-N kg–1 under either ambient CO2 (aCO2; 390 ppm) or elevated CO2 (eCO2; 550 ppm) using free-air CO2 enrichment (SoilFACE). Key Results Under aCO2, field pea biomass was significantly lower at 5 mg NO3–-N kg–1 than at 90 mg NO3–-N kg–1 soil. However, increasing the soil N level significantly reduced nodulation of lateral roots but not the primary root, and nodules were significantly smaller, with 85 % less nodule mass in the 90 NO3–-N kg–1 than in the 5 mg NO3–-N kg–1 treatment, highlighting the inhibitory effects of NO3–. Field pea grown under eCO2 had greater biomass (approx. 30 %) than those grown under aCO2, and was not affected by N level. Overall, the inhibitory effects of NO3– on nodulation and nodule mass appeared to be reduced under eCO2 compared with aCO2, although the effects of CO2 on root growth were not significant. Conclusions Elevated CO2 alleviated the inhibitory effect of soil NO3– on nodulation and N2 fixation and is likely to lead to greater total N content of field pea growing under future elevated CO2 environments. PMID:26346721

  7. Induction of premature mitosis in root meristem cells of Vicia faba and Pisum sativum by various agents is correlated with an increased level of protein phosphorylation.

    PubMed

    Rybaczek, Dorota; Polit, Justyna; Luchniak, Piotr; Maszewski, Janusz

    2002-01-01

    The intra-S-phase checkpoint response to hydroxyurea (HU)-mediated arrest of DNA replication was analysed in root meristems of two legumes, Pisum sativum and Vicia faba. The obtained results suggest that a molecular signal which invokes mechanisms allowing the cells to override the S-M dependency control system may be generated by caffeine (CF) and a number of alternative, yet related chemical agents, benzyl-6-aminopurine (BAP), 2-aminopurine (2-AP), and 6-dimethylaminopurine (DMAP). A variety of aberrant mitotic divisions included chromosomal breaks and gaps, lost and lagging chromatids and chromosomes, acentric fragments, chromosome bridges and micronuclei. Furthermore, similar effects induced by sodium vanadate, an inhibitor of protein phosphatases, extend the number of inhibitors capable of inducing premature chromosome condensation (PCC) in root meristem cells, as well as the range of possible regulatory pathways leading to the transition from S-phase arrest towards abnormal mitosis. Until preprophase, FITC-conjugated monoclonal antibodies (alpha-Y(a)b-FITC) that specifically recognize phosphorylated form of threonine indicate no evident cell cycle-dependent changes in an overall phosphorylation status of root meristem cells in the control plants. Irrespective of the stage of interphase, alpha-Y(p)ab-FITC was localized basically in the cytoplasm, whereas nuclear staining was considerably weaker, with a significant fluorescence confined merely to nucleolar regions. The intensity of alpha-Y(p)ab-FITC staining in HU/CF-treated seedlings was found higher than that in the control plants (with the exception of G2 cells), suggesting a general increase in the level of protein phosphorylation, a physiological response mediated probably by an enhanced activity of the cdc-like protein kinase(s).

  8. Iron absorption from experimental infant formulas based on pea (Pisum sativum)-protein isolate: the effect of phytic acid and ascorbic acid.

    PubMed

    Davidsson, L; Dimitriou, T; Walczyk, T; Hurrell, R F

    2001-01-01

    Infant formula based on pea (Pisum sativum)-protein isolate has been suggested as an alternative to soybean formula in countries where soybean is not a native crop, or when soybean protein cannot be used due to allergic reactions or intolerances. In the present study, Fe absorption from experimental infant formulas based on pea-protein isolate was measured in healthy non-anaemic young women. The influence of phytic acid and ascorbic acid on Fe absorption was evaluated, using a stable-isotope technique based on incorporation of Fe stable-isotope labels into erythrocytes 14 d after administration. Geometric mean Fe absorption increased from 20.7 (+1 SD 41.6, -1 SD 10.3) % to 33.1 (+1 SD 58.6, -1 SD 18.7) %; (P < 0.0001; n 10) after enzymic degradation of virtually all phytic acid. Doubling the molar ratio Fe:ascorbic acid from 1:2.1 to 1:4.2 in the infant formula with native phytic acid content also increased Fe absorption significantly (P < 0.0001; n 10); geometric mean Fe absorption increased from 14.8 (+1 SD 32.1, -1 SD 6.8) % to 22.1 (+1 SD 47.2, -1 SD 10.4) %. These results confirm the inhibitory and enhancing effects of phytic acid and ascorbic acid respectively on Fe absorption, but also indicate relatively high fractional Fe absorption from the pea-protein-based formulas. After adjusting for differences in Fe status, our data indicate that Fe absorption from dephytinised pea protein might be less inhibitory than dephytinised soybean protein as measured in a previous study (Hurrell et al. 1998).

  9. Pyrabactin, an ABA agonist, induced stomatal closure and changes in signalling components of guard cells in abaxial epidermis of Pisum sativum

    PubMed Central

    Puli, Mallikarjuna Rao; Raghavendra, Agepati S.

    2012-01-01

    Pyrabactin, a synthetic agonist of abscisic acid (ABA), inhibits seed germination and hypocotyl growth and stimulates gene expression in a very similar way to ABA, implying the possible modulation of stomatal function by pyrabactin as well. The effect of pyrabactin on stomatal closure and secondary messengers was therefore studied in guard cells of Pisum sativum abaxial epidermis. Pyrabactin caused marked stomatal closure in a pattern similar to ABA. In addition, pyrabactin elevated the levels of reactive oxygen species (ROS), nitric oxide (NO), and cytoplasmic pH levels in guard cells, as indicated by the respective fluorophores. However, apyrabactin, an inactive analogue of ABA, did not affect either stomatal closure or the signalling components of guard cells. The effects of pyrabactin-induced changes were reversed by pharmalogical compounds that modulate ROS, NO or cytoplasmic pH levels, quite similar to ABA effects. Fusicoccin, a fungal toxin, could reverse the stomatal closure caused by pyrabactin, as well as that caused by ABA. Experiments on stomatal closure by varying concentrations of ABA, in the presence of fixed concentration of pyrabactin, and vice versa, revealed that the actions of ABA and pyrabactin were additive. Further kinetic analysis of data revealed that the apparent KD of ABA was increased almost 4-fold in the presence of ABA, suggesting that pyrabactin and ABA were competing with each other either at the same site or close to the active site. It is proposed that pyrabactin could be used to examine the ABA-related signal-transduction components in stomatal guard cells as well as in other plant tissues. It is also suggested that pyrabactin can be used as an antitranspirant or as a priming agent for improving the drought tolerance of crop plants. PMID:22131162

  10. Multiple impacts of the plant growth-promoting rhizobacterium Variovorax paradoxus 5C-2 on nutrient and ABA relations of Pisum sativum

    PubMed Central

    Dodd, Ian C.

    2012-01-01

    Resolving the physiological mechanisms by which rhizobacteria enhance plant growth is difficult, since many such bacteria contain multiple plant growth-promoting properties. To understand further how the 1-aminocyclopropane-1-carboxylate (ACC) deaminase (ACCd)-containing rhizobacterium Variovorax paradoxus 5C-2 affects plant growth, the flows and partitioning of mineral nutrients and abscisic acid (ABA) and ABA metabolism were studied in pea (Pisum sativum) plants following rhizosphere bacterial inoculation. Although root architecture was not affected, inoculation increased root and shoot biomass, and stomatal conductance, by 20, 15, and 24%, respectively, and increased N, P, K, Ca, and Mg uptake by 16, 81, 50, 46, and 58%, respectively. P deposition in inoculated plant roots was 4.9 times higher than that in uninoculated controls. Rhizobacterial inoculation increased root to shoot xylem flows and shoot to root phloem flows of K by 1.8- and 2.1-fold, respectively. In control plants, major sinks for K deposition were the roots and upper shoot (43% and 49% of total uptake, respectively), while rhizobacterial inoculation increased K distribution to the lower shoot at the expense of other compartments (xylem, phloem, and upper shoot). Despite being unable to metabolize ABA in vitro, V. paradoxus 5C-2 decreased root ABA concentrations and accumulation by 40–60%. Although inoculation decreased xylem ABA flows, phloem ABA flows increased. Whether bacterial ACCd attenuates root to shoot ABA signalling requires further investigation, since ABA is critical to maintain growth of droughted plants, and ACCd-containing organisms have been advocated as a means of minimizing growth inhibition of plants in drying soil. PMID:23136167

  11. [Investigation of chromosomes in varieties and translocation lines of pea Pisum sativum L. by FISH, Ag-NOR, and differential DAPI staining].

    PubMed

    Samatadze, T E; Muravenko, O M; Bol'sheva, N L; Amosova, A B; Gostimsckiĭ, S A; Zelenin, A V

    2005-12-01

    The DNA intercalator 9-aminoachridine was used for obtaining high-resolution DAPI patterns of chromosomes of Pisum sativum L. with more than 300 bands per haploid chromosome set. The karyotypes of three pea varieties, Viola, Capital, and Rosa Crown, and two translocation lines, L-108 (T(2-4s)) and M-10 (T(2-7s)), were examined. Based on the results of DAPI staining, we have identified chromosomes, constructed idiograms, and established breakpoints of chromosome translocations. Lines L-108 (T(2-4s)) and M-10 (T(2-7s)) were shown to appear as a result of respectively one translocation between chromosomes 2 and 4 and two translocations between chromosomes 2 and 7. All varieties and translocation lines of pea were examined using fluorescence in situ hybridization (FISH) with telomere repetition probes, 5S and 45S wheat DNA probes. Transcriptional activity of 45S rRNA was detected by Ag-NOR staining. Telomere repetitions were shown to be located only in telomeric chromosome regions. Using high-resolution DAPI staining allowed us to verify localization of 5S genes on pea chromosomes 1, 3, and 5. 45S rDNAs were localized in the secondary constriction regions on the satellite and the satellite thread of chromosome and on the satellite thread and in more proximal satellite heterochromatic region of chromosome 7. The size of 45S rDNA signal on chromosome 7 was larger and its transcriptional activity, higher than the corresponding parameters on chromosome 4 in most of the forms studied. A visual comparison of the results of FISH and Ag-NOR staining of normal and translocated pea chromosomes did not reveal any significant differences between them. The translocations of the satellite chromosomes apparently did not cause significant changes either in the amount of the ribosomal genes or in their transcriptional activity.

  12. Evaluation of the effects of polymeric chitosan/tripolyphosphate and solid lipid nanoparticles on germination of Zea mays, Brassica rapa and Pisum sativum.

    PubMed

    Nakasato, Daniele Y; Pereira, Anderson E S; Oliveira, Jhones L; Oliveira, Halley C; Fraceto, Leonardo F

    2017-08-01

    Although the potential toxicity of many metallic and carbon nanoparticles to plants has been reported, few studies have evaluated the phytotoxic effects of polymeric and solid lipid nanoparticles. The present work described the preparation and characterization of chitosan/tripolyphosphate (CS/TPP) nanoparticles and solid lipid nanoparticles (SLN) and evaluated the effects of different concentrations of these nanoparticles on germination of Zea mays, Brassica rapa, and Pisum sativum. CS/TPP nanoparticles presented an average size of 233.6±12.1nm, polydispersity index (PDI) of 0.30±0.02, and zeta potential of +21.4±1.7mV. SLN showed an average size of 323.25±41.4nm, PDI of 0.23±0.103, and zeta potential of -13.25±3.2mV. Nanotracking analysis enabled determination of concentrations of 1.33×10(10) (CS/TPP) and 3.64×10(12) (SLN) nanoparticles per mL. At high concentrations, CS/TPP nanoparticles caused complete inhibition of germination, and thus negatively affected the initial growth of all tested species. Differently, SLN presented no phytotoxic effects. The different size and composition and the opposite charges of SLN and CS/TPP nanoparticles could be associated with the differential phytotoxicity of these nanomaterials. The present study reports the phytotoxic potential of polymeric CS/TPP nanoparticles towards plants, indicating that further investigation is needed on the effects of such formulations intended for future use in agricultural systems, in order to avoid damage to the environment. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Bean α-amylase inhibitor 1 in transgenic peas (Pisum sativum) provides complete protection from pea weevil (Bruchus pisorum) under field conditions

    PubMed Central

    Morton, Roger L.; Schroeder, Hart E.; Bateman, Kaye S.; Chrispeels, Maarten J.; Armstrong, Eric; Higgins, Thomas J. V.

    2000-01-01

    Two α-amylase inhibitors, called αAI-1 and αAI-2, that share 78% amino acid sequence identity and have a differential specificity toward mammalian and insect α-amylases are present in different accessions of the common bean (Phaseolus vulgaris). Using greenhouse-grown transgenic peas (Pisum sativum), we have shown previously that expression of αAI-1 in pea seeds can provide complete protection against the pea weevil (Bruchus pisorum). Here, we report that αAI-1 also protects peas from the weevil under field conditions. The high degree of protection is explained by our finding that αAI-1 inhibits pea bruchid α-amylase by 80% over a broad pH range (pH 4.5–6.5). αAI-2, on the other hand, is a much less effective inhibitor of pea bruchid α-amylase, inhibiting the enzyme by only 40%, and only in the pH 4.0–4.5 range. Nevertheless, this inhibitor was still partially effective in protecting field-grown transgenic peas against pea weevils. The primary effect of αAI-2 appeared to be a delay in the maturation of the larvae. This contrasts with the effect of αAI-1, which results in larval mortality at the first or second instar. These results are discussed in relationship to the use of amylase inhibitors with different specificities to bring about protection of crops from their insect pests or to decrease insect pest populations below the economic injury level. PMID:10759552

  14. Integrated effect of microbial antagonist, organic amendment and fungicide in controlling seedling mortality (Rhizoctonia solani) and improving yield in pea (Pisum sativum L.).

    PubMed

    Akhter, Wasira; Bhuiyan, Mohamed Khurshed Alam; Sultana, Farjana; Hossain, Mohamed Motaher

    2015-01-01

    The study evaluated the comparative performance of a few microbial antagonists, organic amendments and fungicides and their integration for the management of seedling mortality (Rhizoctonia solani Kühn) and yield improvement in pea (Pisum sativum L.). Before setting the experiment in field microplots, a series of in vitro and in vivo experiments were conducted to select a virulent isolate of R. solani, an effective antagonistic isolate of Trichoderma harzianum, a fungitoxic organic amendment and an appropriate fungicide. A greenhouse pathogenicity test compared differences in seedling mortality in pea inoculated by four isolates of R. solani and identified the isolate RS10 as the most virulent one. Among the 20 isolates screened in dual culture assay on PDA, T. harzianum isolate T-3 was found to show the highest (77.22%) inhibition of the radial growth of R. solani. A complete inhibition (100.00%) of colony growth of R. solani was observed when fungicide Bavistin 50 WP and Provax-200 at the rate of 100 and 250 ppm, respectively, were used, while Provax-200 was found to be highly compatible with T. harzianum. Mustard oilcake gave maximum inhibition (60.28%) of the radial growth of R. solani at all ratios, followed by sesame oilcake and tea waste. Integration of soil treatment with T. harzianum isolate T-3 and mustard oilcake and seed treatment with Provax-200 appeared to be significantly superior in reducing seedling mortality and improving seed yield in pea in comparison to any single or dual application of them in the experimental field. The research results will help growers develop integrated disease management strategies for the control of Rhizoctonia disease in pea. The research results show the need for an integrating selective microbial antagonist, organic amendment and fungicide to achieve appropriate management of seedling mortality (R. solani) and increase of seed yield in pea.

  15. Stomatal response and leaf injury of Pisum sativum L. with SO/sub 2/ and O/sub 3/ exposures. I. Influence of pollutant level and leaf maturity

    SciTech Connect

    Olszyk, D.M.; Tibbitts, T.W.

    1981-03-01

    Plants of Pisum sativum L. Alsweet were grown under a controlled environment and exposed to SO/sub 2/ and O/sub 3/ to determine whether changes in stomatal aperture during exposure were related to subsequent leaf injury. Stomata consistently closed with injurious levels of SO/sub 2/ and O/sub 3/. Measurements with diffusion porometers demonstrated approx. = 75 and 25% lower conductance with SO/sub 2/ and O/sub 3/ exposures, respectively, compared to the conductance of control plants. Stomata also showed a closing response with noninjurious levels of SO/sub 2/ but an opening response with noninjurious levels of O/sub 3/. Stomata closed to the same degree with combinations of SO/sub 2/ plus O/sub 3/ as with SO/sub 2/ alone. Stomata of expanding leaves closed more during pollutant exposures than stomata of expanded leaves. The abaxial and adaxial stomata both exhibited closure with SO/sub 2/ and combinations of SO/sub 2/ plus O/sub 3/, but abaxial stomata tended to close and adaxial stomata tended to open with exposure to O/sub 3/ alone. The changes in stomatal aperture were not closely correlated with the amount of leaf injury produced by different pollutant levels. Stomata closed, not only with exposure to pollutant levels that caused severe necrosis, but also with levels that caused only a trace of injury. There was no evidence of a reduced amount of closure or even stomatal opening with combinations of SO/sub 2/ and O/sub 3/ compared to plants exposed to the pollutants alone to explain the large amount of injury to plants exposed to pollutant combinations.

  16. Studies on the Control of Ascochyta Blight in Field Peas (Pisum sativum L.) Caused by Ascochyta pinodes in Zhejiang Province, China.

    PubMed

    Liu, Na; Xu, Shengchun; Yao, Xiefeng; Zhang, Guwen; Mao, Weihua; Hu, Qizan; Feng, Zhijuan; Gong, Yaming

    2016-01-01

    Ascochyta blight, an infection caused by a complex of Ascochyta pinodes, Ascochyta pinodella, Ascochyta pisi, and/or Phoma koolunga, is a destructive disease in many field peas (Pisum sativum L.)-growing regions, and it causes significant losses in grain yield. To understand the composition of fungi associated with this disease in Zhejiang Province, China, a total of 65 single-pycnidiospore fungal isolates were obtained from diseased pea samples collected from 5 locations in this region. These isolates were identified as Ascochyta pinodes by molecular techniques and their morphological and physiological characteristics. The mycelia of ZJ-1 could penetrate pea leaves across the stomas, and formed specific penetration structures and directly pierced leaves. The resistance level of 23 available pea cultivars was tested against their representative isolate A. pinodes ZJ-1 using the excised leaf-assay technique. The ZJ-1 mycelia could penetrate the leaves of all tested cultivars, and they developed typical symptoms, which suggested that all tested cultivars were susceptible to the fungus. Chemical fungicides and biological control agents were screened for management of this disease, and their efficacies were further determined. Most of the tested fungicides (11 out of 14) showed high activity toward ZJ-1 with EC50 < 5 μg/mL. Moreover, fungicides, including tebuconazole, boscalid, iprodione, carbendazim, and fludioxonil, displayed more than 80% disease control efficacy under the recorded conditions. Three biocontrol strains of Bacillus sp. and one of Pantoea agglomerans were isolated from pea-related niches and significantly reduced the severity of disease under greenhouse and field conditions. To our knowledge, this is the first study on ascochyta blight in field peas, and results presented here will be useful for controlling the disease in this area.

  17. De novo transcriptome assembly reveals high transcriptional complexity in Pisum sativum axillary buds and shows rapid changes in expression of diurnally regulated genes.

    PubMed

    Kerr, Stephanie C; Gaiti, Federico; Beveridge, Christine A; Tanurdzic, Milos

    2017-03-02

    The decision for a bud to grow into a branch is a key regulatory process affecting plant architecture. In order to study molecular processes regulating axillary bud outgrowth in the model plant garden pea (Pisum sativum), we sequenced the axillary bud transcriptome and performed de novo transcriptome assembly. We assembled a pea axillary bud transcriptome into 81,774 transcripts comprised of 194,067 isoforms. This new pea transcriptome resource is both comprehensive and representative, as shown by comparison to other available pea sequence resources. Over half of the transcriptome could be annotated based on sequence homology to Arabidopsis thaliana proteins, while almost one quarter of the isoforms were identified as putative long non-coding RNAs (lncRNAs). This transcriptome will be useful in studies of pea buds because it includes genes expressed specifically in buds which are not represented in other transcriptome studies. We also investigated the impact of a short time collection series on gene expression. Differential gene expression analysis identified 142 transcripts changing within the short 170 min time frame that the buds were harvested within. Thirty-three of these transcripts are implicated in diurnal fluctuations in other flowering plants, while the remaining transcripts include 31 putative lncRNA. Further investigation of the differentially expressed transcripts found an enrichment of genes involved in post-transcriptional regulation, including RNA processing and modification, as well as genes involved in fatty acid biosynthesis and oxidative phosphorylation. We have sequenced and assembled a high quality pea bud transcriptome containing both coding and non-coding RNA transcripts that will be useful for further studies into axillary bud outgrowth. Over the short sample collection time frame of just 170 min, we identified differentially expressed coding and non-coding RNA, some of which are implicated in diurnal regulation, highlighting the utility

  18. Ascochyta blight disease of pea (Pisum sativum L.): defence-related candidate genes associated with QTL regions and identification of epistatic QTL.

    PubMed

    Timmerman-Vaughan, Gail M; Moya, Leire; Frew, Tonya J; Murray, Sarah R; Crowhurst, Ross

    2016-05-01

    Advances have been made in our understanding of Ascochyta blight resistance genetics through mapping candidate genes associated with QTL regions and demonstrating the importance of epistatic interactions in determining resistance. Ascochyta blight disease of pea (Pisum sativum L.) is economically significant with worldwide distribution. The causal pathogens are Didymella pinodes, Phoma medicaginis var pinodella and, in South Australia, P. koolunga. This study aimed to identify candidate genes that map to quantitative trait loci (QTL) for Ascochyta blight field disease resistance and to explore the role of epistatic interactions. Candidate genes associated with QTL were identified beginning with 101 defence-related genes from the published literature. Synteny between pea and Medicago truncatula was used to narrow down the candidates for mapping. Fourteen pea candidate sequences were mapped in two QTL mapping populations, A26 × Rovar and A88 × Rovar. QTL peaks, or the intervals containing QTL peaks, for the Asc2.1, Asc4.2, Asc4.3 and Asc7.1 QTL were defined by four of these candidate genes, while another three candidate genes occurred within 1.0 LOD confidence intervals. Epistasis involving QTL × background marker and background marker × background marker interactions contributed to the disease response phenotypes observed in the two mapping populations. For each population, five pairwise interactions exceeded the 5% false discovery rate threshold. Two candidate genes were involved in significant pairwise interactions. Markers in three genomic regions were involved in two or more epistatic interactions. Therefore, this study has identified pea defence-related sequences that are candidates for resistance determination, and that may be useful for marker-assisted selection. The demonstration of epistasis informs breeders that the architecture of this complex quantitative resistance includes epistatic interactions with non-additive effects.

  19. Automated integrative high-throughput phenotyping of plant shoots: a case study of the cold-tolerance of pea (Pisum sativum L.).

    PubMed

    Humplík, Jan F; Lazár, Dušan; Fürst, Tomáš; Husičková, Alexandra; Hýbl, Miroslav; Spíchal, Lukáš

    2015-01-01

    Recently emerging approaches to high-throughput plant phenotyping have discovered their importance as tools in unravelling the complex questions of plant growth, development and response to the environment, both in basic and applied science. High-throughput methods have been also used to study plant responses to various types of biotic and abiotic stresses (drought, heat, salinity, nutrient-starving, UV light) but only rarely to cold tolerance. We present here an experimental procedure of integrative high-throughput in-house phenotyping of plant shoots employing automated simultaneous analyses of shoot biomass and photosystem II efficiency to study the cold tolerance of pea (Pisum sativum L.). For this purpose, we developed new software for automatic RGB image analysis, evaluated various parameters of chlorophyll fluorescence obtained from kinetic chlorophyll fluorescence imaging, and performed an experiment in which the growth and photosynthetic activity of two different pea cultivars were followed during cold acclimation. The data obtained from the automated RGB imaging were validated through correlation of pixel based shoot area with measurement of the shoot fresh weight. Further, data obtained from automated chlorophyll fluorescence imaging analysis were compared with chlorophyll fluorescence parameters measured by a non-imaging chlorophyll fluorometer. In both cases, high correlation was obtained, confirming the reliability of the procedure described. This study of the response of two pea cultivars to cold stress confirmed that our procedure may have important application, not only for selection of cold-sensitive/tolerant varieties of pea, but also for studies of plant cold-response strategies in general. The approach, provides a very broad tool for the morphological and physiological selection of parameters which correspond to shoot growth and the efficiency of photosystem II, and is thus applicable in studies of various plant species and crops.

  20. The photorespiratory hydrogen shuttle. Synthesis of phthalonic acid and its use in the characterization of the malate/aspartate shuttle in pea (Pisum sativum) leaf mitochondria.

    PubMed Central

    Dry, I B; Dimitriadis, E; Ward, A D; Wiskich, J T

    1987-01-01

    A method is presented for the preparation of pure phthalonic acid (PTA) in high yields. This PTA was used to determine the capacity of the malate/aspartate shuttle in pea (Pisum sativum) leaf mitochondria. The inhibition of glycine-dependent O2 uptake in the combined presence of 5 mM-aspartate and 5 mM-2-oxoglutarate (2-OG) was decreased by 55 +/- 22% (n = 13) in washed and 50 +/- 2% (n = 11) in purified mitochondria by 0.23 mM-PTA. This concentration of PTA had no effect on the oxidation of 5 mM-2-OG, suggesting that part of the observed inhibition of O2 uptake in the presence of aspartate and 2-OG was due to the production of oxaloacetate (OAA) by aspartate aminotransferase external to the mitochondrial inner membrane. Levels of external aspartate aminotransferase were estimated to be 24 +/- 1% (n = 4) and 13 +/- 1% (n = 4) of the total mitochondrial activity in washed and purified mitochondria respectively. Malate/aspartate-shuttle activity was estimated directly by measuring rates of malate efflux from isolated mitochondria and was found to match estimates of shuttle activity based on the PTA-insensitive inhibition of O2 uptake. Comparisons of malate/aspartate- and malate/OAA-shuttle activities indicated potentially similar rates of NADH export from pea leaf mitochondria under conditions in vivo. These extrapolated to whole-tissue rates of 5-11 mumol of NADH.h-1.mg of chlorophyll-1. The potential role of the malate/aspartate shuttle in the support of photorespiratory glycine oxidation in leaf tissue is discussed. PMID:3663185

  1. Cr(VI) induces DNA damage, cell cycle arrest and polyploidization: a flow cytometric and comet assay study in Pisum sativum.

    PubMed

    Rodriguez, Eleazar; Azevedo, Raquel; Fernandes, Pedro; Santos, Conceição

    2011-07-18

    Chromium(VI) is recognized as the most toxic valency of Cr, but its genotoxicity and cytostaticity in plants is still poorly studied. In order to analyze Cr(VI) cyto- and gentotoxicity, Pisum sativum L. plants were grown in soil and watered with solutions with different concentrations of Cr up to 2000 mg/L. After 28 days of exposure, leaves showed no significant variations in either cell cycle dynamics or ploidy level. As for DNA damage, flow cytometric (FCM) histograms showed significant differences in full peak coefficient of variation (FPCV) values, suggesting clastogenicity. This is paralleled by the Comet assay results, showing an increase in DNA damage for 1000 and 2000 mg/L. In roots, exposure to 2000 mg/L resulted in cell cycle arrest at the G(2)/M checkpoint. It was also verified that under the same conditions 40% of the individuals analyzed suffered polyploidization having both 2C and 4C levels. DNA damage analysis by the Comet assay and FCM revealed dose-dependent increases in DNA damage and FPCV. Through this, we have unequivocally demonstrated for the first time in plants that Cr exposure can result in DNA damage, cell cycle arrest, and polyploidization. Moreover, we critically compare the validity of the Comet assay and FCM in evaluating cytogenetic toxicity tests in plants and demonstrate that the data provided by both techniques complement each other and present high correlation levels. In conclusion, the data presented provides new insight on Cr effects in plants in general and supports the use of the parameters tested in this study as reliable endpoints for this metal toxicity in plants. © 2011 American Chemical Society

  2. GH3 expression and IAA-amide synthetase activity in pea (Pisum sativum L.) seedlings are regulated by light, plant hormones and auxinic herbicides.

    PubMed

    Ostrowski, Maciej; Jakubowska, Anna

    2013-03-01

    The formation of auxin conjugates is one of the important regulatory mechanisms for modulating IAA action. Several auxin-responsive GH3 genes encode IAA-amide synthetases that are involved in the maintenance of hormonal homeostasis by conjugating excess IAA to amino acids. Recently, the data have revealed novel regulatory functions of several GH3 proteins in plant growth, organ development, fruit ripening, light signaling, abiotic stress tolerance and plant defense responses. Indole-3-acetyl-aspartate (IAA-Asp) synthetase catalyzing IAA conjugation to aspartic acid in immature seeds of pea (Pisum sativum L.) was purified and characterized during our previous investigations. In this study, we examined the effect of auxin and other plant hormones (ABA, GA, kinetin, JA, MeJA, SA), different light conditions (red, far-red, blue, white light), and auxinic herbicides (2,4-D, Dicamba, Picloram) on the expression of a putative GH3 gene and IAA-amide synthesizing activity in 10-d-old pea seedlings. Quantitative RT-PCR analysis indicated that the PsGH3-5 gene, weakly expressed in control sample, was visibly induced in response to all plant hormones, different light wavelengths and the auxinic herbicides tested. Protein A immunoprecipitation/gel blot analysis using anti-AtGH3.5 antibodies revealed a similar pattern of changes on the protein levels in response to all treatments. IAA-amide synthetase activity determined with aspartate as a substrate, not detectable in control seedlings, was positively affected by a majority of treatments. Based on these results, we suggest that PsGH3-5 may control the growth and development of pea plants in a way similar to the known GH3 genes from other plant species. Copyright © 2012 Elsevier GmbH. All rights reserved.

  3. Growth performance and carcass characteristics of guinea fowl broilers fed micronized-dehulled pea (Pisum sativum L.) as a substitute for soybean meal.

    PubMed

    Laudadio, V; Nahashon, S N; Tufarelli, V

    2012-11-01

    This study was conducted to evaluate the effect of substitution of soybean meal (SBM) with dehulled-micronized peas (Pisum sativum) in diets of guinea fowl broilers on their growth performance, carcass yields, and fatty acid composition of meat. One hundred forty 1-d-old guinea fowl keets were randomly assigned to 2 dietary treatments, which were fed from hatch to 12 wk. The birds were fed 2 wheat middling-based diets comprising a control diet, which contained SBM (78 g/kg) and a test diet containing dehulled-micronized peas (180 g/kg) as the main protein source. The substitution of SBM with peas had no adverse effect on growth performance, dressing percentage, or breast and thigh muscle relative weights of the guinea broilers. However, a reduction of abdominal fat content (P < 0.05) was observed in birds fed the pea diet compared with the control. Breast and thigh meat of birds fed the pea diet had higher lightness scores (P < 0.05) and water-holding capacity (P < 0.01) than the control. Meat from guinea fowls fed the pea diet had less cholesterol (P < 0.01) and lipids (P < 0.05), and higher concentrations of phospholipids (P < 0.05). Feeding peas increased polyunsaturated fatty acid concentration in breast and thigh muscles, and decreased the saturated fatty acid concentration. Feeding the pea diet also lowered the n-6/n-3 polyunsaturated fatty acid ratio of the guinea broiler muscles. Our results suggest that replacing the conventional SBM as the protein source with dehulled-micronized pea meal in diets of guinea fowls broilers can improve carcass quality and favorable lipid profile without adversely affecting growth performance traits.

  4. Changes in the protein patterns in pea (Pisum sativum L.) roots under the influence of long- and short-term chilling stress and post-stress recovery.

    PubMed

    Badowiec, Anna; Swigonska, Sylwia; Weidner, Stanisław

    2013-10-01

    Amongst many factors restricting geographical distribution of plants and crop productivity, low temperature is one of the most important. To gain better understanding of the molecular response of germinating pea (Pisum sativum L.) to low temperature, we investigated the influence of long and short chilling stress as well as post-stress recovery on the alterations in the root proteomes. The impact of long stress was examined on the pea seeds germinating in the continuous chilling conditions of 10 °C for 8 days (LS). To examine the impact of short stress, pea seeds germinating for 72 h in the optimal temperature of 20 °C were subjected to 24-h chilling (SS). Additionally, both stress treatments were followed by 24 h of recovery in the optimal conditions (accordingly LSR and SR). Using the 2D gel electrophoresis and MALDI-TOF MS protein identification, it was revealed, that most of the proteins undergoing regulation under the applied conditions were implicated in metabolism, protection against stress, cell cycle regulation, cell structure maintenance and hormone synthesis, which altogether may influence root growth and development in the early stages of plant life. The obtained results have shown that most of detected alterations in the proteome patterns of pea roots are dependent on stress duration. However, there are some analogical response pathways which are triggered regardless of stress length. The functions of proteins which accumulation has been changed by chilling stress and post-stress recovery are discussed here in relation to their impact on pea roots development.

  5. The Relationship between H2 Evolution and Acetylene Reduction in Pisum sativum-Rhizobium leguminosarum Symbioses Differing in Uptake Hydrogenase Activity 1

    PubMed Central

    Mahon, John D.; Nelson, Louise M.

    1986-01-01

    Peas (Pisum sativum L.) were inoculated with strains of Rhizobium leguminosarum having different levels of uptake hydrogenase (Hup) activity and were grown in sterile Leonard jars under controlled conditions. Rates of H2 evolution and acetylene reduction were determined for intact nodulated roots at intervals after the onset of darkness or after removal of the shoots. Hup activity was estimated using treatment plants or equivalent plants from the growth chamber, by measuring the uptake of H2 or 3H2 in the presence of acetylene. In all cases, the rate of H2 evolution was a continuous function of the rate of acetylene reduction. In symbioses with no demonstrable Hup activity, H2 evolution increased in direct proportion to acetylene reduction and the slopes were similar with the Hup− strains NA502 and 128C79. Hup activity was similar in strains 128C30 and 128C52 but significantly lower in strain 128C54. With these strains, the slopes of the H2 evolution versus acetylene reduction curves initially increased with acetylene reduction, but became constant and similar to those for the Hup− strains at high rates of acetylene reduction. On these parallel portions of the curves, the decreases in H2 evolution by Hup+ strains were similar in magnitude to their H2-saturated rates of Hup activity. The curvilinear relationship between H2 evolution and acetylene reduction for a representative Hup+ strain (128C52) was the same, regardless of the experimental conditions used to vary the nitrogenase activity. PMID:16664984

  6. Construction and characterization of two bacterial artificial chromosome libraries of pea (Pisum sativum L.) for the isolation of economically important genes.

    PubMed

    Coyne, C J; McClendon, M T; Walling, J G; Timmerman-Vaughan, G M; Murray, S; Meksem, K; Lightfoot, D A; Shultz, J L; Keller, K E; Martin, R R; Inglis, D A; Rajesh, P N; McPhee, K E; Weeden, N F; Grusak, M A; Li, C-M; Storlie, E W

    2007-09-01

    Pea (Pisum sativum L.) has a genome of about 4 Gb that appears to share conserved synteny with model legumes having genomes of 0.2-0.4 Gb despite extensive intergenic expansion. Pea plant inventory (PI) accession 269818 has been used to introgress genetic diversity into the cultivated germplasm pool. The aim here was to develop pea bacterial artificial chromosome (BAC) libraries that would enable the isolation of genes involved in plant disease resistance or control of economically important traits. The BAC libraries encompassed about 3.2 haploid genome equivalents consisting of partially HindIII-digested DNA fragments with a mean size of 105 kb that were inserted in 1 of 2 vectors. The low-copy oriT-based T-DNA vector (pCLD04541) library contained 55 680 clones. The single-copy oriS-based vector (pIndigoBAC-5) library contained 65 280 clones. Colony hybridization of a universal chloroplast probe indicated that about 1% of clones in the libraries were of chloroplast origin. The presence of about 0.1% empty vectors was inferred by white/blue colony plate counts. The usefulness of the libraries was tested by 2 replicated methods. First, high-density filters were probed with low copy number sequences. Second, BAC plate-pool DNA was used successfully to PCR amplify 7 of 9 published pea resistance gene analogs (RGAs) and several other low copy number pea sequences. Individual BAC clones encoding specific sequences were identified. Therefore, the HindIII BAC libraries of pea, based on germplasm accession PI 269818, will be useful for the isolation of genes underlying disease resistance and other economically important traits.

  7. Response of Pisum sativum (Fabales: Fabaceae) to Sitona lineatus (Coleoptera: Curculionidae) infestation: effect of adult weevil density on damage, larval population, and yield loss.

    PubMed

    Vankosky, M A; Cárcamo, H A; Dosdall, L M

    2011-10-01

    Sitona lineatus L. (Coleoptera: Curculionidae) is an invasive pest in North America and its geographical range is currently expanding across the Canadian prairies. Adults and larvae of S. lineatus feed upon the foliage and root nodules, respectively, of field pea, Pisum sativum L. (Fabales: Fabaceae), and may contribute to economic losses when population densities are high. Integrated pest management (IPM) programs that incorporate economic thresholds should be used to manage S. lineatus populations in a sustainable manner. The impact of nitrogen fertilizer on field pea yield and the relationships between adult weevil density and above- and below-ground damage and yield were investigated in southern Alberta, Canada using exclusion cages on field pea plots. In each cage, 32 field pea plants were exposed to weevil densities ranging from zero to one adult weevil per plant. Nitrogen-fertilized plants yielded 16% more than unfertilized plants. Nitrogen-fertilized plants had fewer root nodules than unfertilized plants, but fertilizer had no effect on foliar feeding by S. lineatus. Adult density affected foliar feeding damage, with increases in above-ground damage associated with increases in S. lineatus density. Adult density did not affect root nodule damage, larval density, foliar biomass or seed weight. Overall, these results indicate that terminal leaf damage may be used to estimate adult weevil density but cannot be used to predict larval density or yield loss. Further research is required to better understand the impact of larval damage on yield and determine if economic thresholds can be developed using data from large-scale production systems.

  8. Remote Sensing Study of the Influence of Different Herbicides on the Leaf Spectral Reflectance and Fluorescence of Pea Plants (Pisum sativum L.)

    NASA Astrophysics Data System (ADS)

    Krezhova, Dora; Yanev, Tony; Iliev, Ilko; Alexieva, Vera; Tsaneva, Mariana

    The effective use of airborne and satellite-based remote sensor systems in resource management, agriculture, mineral exploration and environmental monitoring requires an understanding of the nature and limitations of the high-resolution remote sensing data and of various strategies for processing and interpreting it. In developing the necessary knowledge base, ground-based measurements are the expedient source of information. In this study, remote sensing techniques were applied in laboratory for detection of the influence of herbicides 2.4-D, glyphosate, fluridone and acifluorfen on the leaf spectral reflectance and fluorescence of pea plants (Pisum sativum L.). According to the classification of the Herbicide Resistance Action Committee with reference to their mode of action they belong to different groups: synthetic auxins - O (2.4-D), inhibition of EPSP synthase - G (glyphosate), photobleaching - F1 (fluridone), and inhibition of PPO - E (acifluorfen). During the last 40 years, these herbicides are among the ones used most widely in agriculture worldwide. The plants studied were grown hydroponically in a growth chamber in a nutritious medium to which every herbicide was added at two low concentrations (1 µM, 0.1 µM) with respect to the field dose applied in the agricultural practice. High-resolution spectral data for leaf spectral reflectance and fluorescence were collected from freshly detached leaves using three multichannel spectrometers. Spectral reflectance characteristics were obtained from the leaf reflectance referenced against a standard (white diffuse screen) in the visible and near infrared ranges of the electromagnetic spectrum (450÷850 nm). Fluorescence spectra were taken in the spectral range 650-850 nm. To assess the changes arising in leaf spectral reflectance under the herbicide action we developed and applied an analytical approach based on discriminant analysis and other statistical methods. The spectral characteristics were analyzed in

  9. Free-air CO2 enrichment (FACE) reduces the inhibitory effect of soil nitrate on N2 fixation of Pisum sativum.

    PubMed

    Butterly, Clayton R; Armstrong, Roger; Chen, Deli; Tang, Caixian

    2016-01-01

    Additional carbohydrate supply resulting from enhanced photosynthesis under predicted future elevated CO2 is likely to increase symbiotic nitrogen (N) fixation in legumes. This study examined the interactive effects of atmospheric CO2 and nitrate (NO3(-)) concentration on the growth, nodulation and N fixation of field pea (Pisum sativum) in a semi-arid cropping system. Field pea was grown for 15 weeks in a Vertosol containing 5, 25, 50 or 90 mg NO3(-)-N kg(-1) under either ambient CO2 (aCO2; 390 ppm) or elevated CO2 (eCO2; 550 ppm) using free-air CO2 enrichment (SoilFACE). Under aCO2, field pea biomass was significantly lower at 5 mg NO3(-)-N kg(-1) than at 90 mg NO3(-)-N kg(-1) soil. However, increasing the soil N level significantly reduced nodulation of lateral roots but not the primary root, and nodules were significantly smaller, with 85% less nodule mass in the 90 NO3(-)-N kg(-1) than in the 5 mg NO3(-)-N kg(-1) treatment, highlighting the inhibitory effects of NO3(-). Field pea grown under eCO2 had greater biomass (approx. 30%) than those grown under aCO2, and was not affected by N level. Overall, the inhibitory effects of NO3(-) on nodulation and nodule mass appeared to be reduced under eCO2 compared with aCO2, although the effects of CO2 on root growth were not significant. Elevated CO2 alleviated the inhibitory effect of soil NO3(-) on nodulation and N2 fixation and is likely to lead to greater total N content of field pea growing under future elevated CO2 environments. © The Author 2015. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  10. Molecular basis of processing-induced changes in protein structure in relation to intestinal digestion in yellow and green type pea (Pisum sativum L.): A molecular spectroscopic analysis.

    PubMed

    Yu, Gloria Qingyu; Warkentin, Tom; Niu, Zhiyuan; Khan, Nazir A; Yu, Peiqiang

    2015-12-05

    The objectives of this study were (1) to quantify the protein inherent molecular structural features of green cotyledon (CDC Striker) and yellow cotyledon (CDC Meadow) pea (Pisum sativum L.) seeds using molecular spectroscopic technique (FT/IR-ATR); (2) measure the denaturation of protein molecular makeup in the two types of pea during dry roasting (120°C for 60 min), autoclaving (120°C for 60 min) or microwaving (for 5 min); and (3) correlate the heat-induced changes in protein molecular makeup to the corresponding changes in protein digestibility determined using modified three-step in vitro procedure. Compared with yellow-type, the green-type peas had higher (P<0.05) ratios of amide I to II peak height (1.698 vs. 1.805) and area (1.843 vs. 2.017). A significant correlation was observed between the amide I and II peak height (r=0.48) and peak area (r=-0.42) ratio with protein content. Compared with yellow-type, the green-type peas had lower (P<0.05) α-helix:β-sheet ratio (1.015 vs. 0.926), indicating varietal difference in protein secondary structure makeup. All processing applications increased α-helix:β-sheet ratio, with the largest (P<0.05) increase being observed with roasting and microwaving. The heat-induced changes in α-helix:β-sheet ratio was strongly correlated to intestinal digestibility of protein within the green (r=-0. 86) and yellow (r=0.81) pea-types. However, across the pea types the correlation was not significant. Principal component and hierarchical cluster analyses on the entire spectral data from the amide region (ca. 1727-1480 cm(-1)) were able to visualize and discriminate the structural difference between pea varieties and processing treatments. This study shows that the molecular spectroscopy can be used as a rapid tool to screen the protein value of raw and heat-treated peas. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Proteomics offers insight to the mechanism behind Pisum sativum L. response to pea seed-borne mosaic virus (PSbMV).

    PubMed

    Cerna, Hana; Černý, Martin; Habánová, Hana; Šafářová, Dana; Abushamsiya, Kifah; Navrátil, Milan; Brzobohatý, Břetislav

    2017-02-05

    Pea seed-borne mosaic virus (PSbMV) significantly reduces yields in a broad spectra of legumes. The eukaryotic translation initiation factor has been shown to confer resistance to this pathogen, thus implying that translation and proteome dynamics play a role in resistance. This study presents the results of a proteome-wide analysis of Pisum sativum L. response to PSbMV infection. LC-MS profiling of two contrasting pea cultivars, resistant (B99) and susceptible (Raman) to PSbMV infection, detected >2300 proteins, 116 of which responded to PSbMV ten and/or twenty days post-inoculation. These differentially abundant proteins are involved in number of processes that have previously been reported in the plant-pathogen response, including protein and amino acid metabolism, stress signaling, redox homeostasis, carbohydrate metabolism, and lipid metabolism. We complemented our proteome-wide analysis work with targeted analyses of free amino acids and selected small molecules, fatty acid profiling, and enzyme activity assays. Data from these additional experiments support our findings and validate the biological relevance of the observed proteome changes. We found surprising similarities in the resistant and susceptible cultivars, which implies that a seemingly unaffected plant, with no detectable levels of PSbMV, actively suppresses viral replication. Plant resistance to PSbMV is connected to translation initiation factors, yet the processes involved are still poorly understood at the proteome level. To the best of our knowledge, this is the first survey of the global proteomic response to PSbMV in plants. The combination of label-free LC-MS profiling and two contrasting cultivars (resistant and susceptible) provided highly sensitive snapshots of protein abundance in response to PSbMV infection. PSbMV is a member of the largest family of plant viruses and our results are in accordance with previously characterized potyvirus-responsive proteomes. Hence, the results of this

  12. Expression of Pisum sativum PsAO3 gene, which encodes an aldehyde oxidase utilizing abscisic aldehyde, is induced under progressively but not rapidly imposed drought stress.

    PubMed

    Zdunek-Zastocka, Edyta; Sobczak, Mirosław

    2013-10-01

    Aldehyde oxidase (AO; EC 1.2.3.1) catalyzes the final step of abscisic acid (ABA) biosynthesis, which is the oxidation of abscisic aldehyde (ABAld) to ABA. Gene expression analyses indicate that the stress-induced Pisum sativum PsAOγ isoform, which effectively uses ABAld as a substrate, is encoded by the PsAO3 gene. PsAO3 was heterologously expressed in Pichia pastoris and the recombinant PsAO3 protein revealed substrate preferences highly similar to the native PsAOγ protein present in the pea leaves and roots. Both proteins prefer indole-3-aldehyde and naphthaldehyde as substrates, although high activities against abscisic aldehyde and citral were also observed. The Km values of PsAO3 for naphthaldehyde and abscisic aldehyde (4.6 and 5.1 μM, respectively) were the lowest among the substrates tested. PsAO3 activity was almost completely inhibited by potassium cyanide, diphenyleneiodonium, and methanol. Rapidly imposed drought stress did not increase the level of PsAO3 mRNA or activity of any AO isoform, although an enhanced ABA accumulation and induction of PsNCED2 and -3 (9-cis-epoxycarotenoid dioxygenase; EC 1.13.11.51) expression, both in the pea roots and leaves, was observed. During a progressively induced drought, the level of PsAO3 transcript and PsAOγ activity increased significantly in the roots and leaves, whereas ABA accumulation occurred only in the leaves where it was accompanied by induction of the PsNCED3 expression. Therefore, we suppose that next to NCED, also AO (mainly PsAOγ) might be involved in regulation of the drought-induced ABA synthesis. However, while the "constitutive activity" of PsAOγ is sufficient for the fast generation of ABA under rapid drought stress, the enhanced PsAOγ activity is required for the progressive and long-term ABA accumulation in the leaves under progressive drought stress. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  13. Repetitive DNA in the pea (Pisum sativum L.) genome: comprehensive characterization using 454 sequencing and comparison to soybean and Medicago truncatula

    PubMed Central

    Macas, Jiří; Neumann, Pavel; Navrátilová, Alice

    2007-01-01

    Background Extraordinary size variation of higher plant nuclear genomes is in large part caused by differences in accumulation of repetitive DNA. This makes repetitive DNA of great interest for studying the molecular mechanisms shaping architecture and function of complex plant genomes. However, due to methodological constraints of conventional cloning and sequencing, a global description of repeat composition is available for only a very limited number of higher plants. In order to provide further data required for investigating evolutionary patterns of repeated DNA within and between species, we used a novel approach based on massive parallel sequencing which allowed a comprehensive repeat characterization in our model species, garden pea (Pisum sativum). Results Analysis of 33.3 Mb sequence data resulted in quantification and partial sequence reconstruction of major repeat families occurring in the pea genome with at least thousands of copies. Our results showed that the pea genome is dominated by LTR-retrotransposons, estimated at 140,000 copies/1C. Ty3/gypsy elements are less diverse and accumulated to higher copy numbers than Ty1/copia. This is in part due to a large population of Ogre-like retrotransposons which alone make up over 20% of the genome. In addition to numerous types of mobile elements, we have discovered a set of novel satellite repeats and two additional variants of telomeric sequences. Comparative genome analysis revealed that there are only a few repeat sequences conserved between pea and soybean genomes. On the other hand, all major families of pea mobile elements are well represented in M. truncatula. Conclusion We have demonstrated that even in a species with a relatively large genome like pea, where a single 454-sequencing run provided only 0.77% coverage, the generated sequences were sufficient to reconstruct and analyze major repeat families corresponding to a total of 35–48% of the genome. These data provide a starting point for

  14. Genome-wide SNP identification, linkage map construction and QTL mapping for seed mineral concentrations and contents in pea (Pisum sativum L.).

    PubMed

    Ma, Yu; Coyne, Clarice J; Grusak, Michael A; Mazourek, Michael; Cheng, Peng; Main, Dorrie; McGee, Rebecca J

    2017-02-13

    Marker-assisted breeding is now routinely used in major crops to facilitate more efficient cultivar improvement. This has been significantly enabled by the use of next-generation sequencing technology to identify loci and markers associated with traits of interest. While rich in a range of nutritional components, such as protein, mineral nutrients, carbohydrates and several vitamins, pea (Pisum sativum L.), one of the oldest domesticated crops in the world, remains behind many other crops in the availability of genomic and genetic resources. To further improve mineral nutrient levels in pea seeds requires the development of genome-wide tools. The objectives of this research were to develop these tools by: identifying genome-wide single nucleotide polymorphisms (SNPs) using genotyping by sequencing (GBS); constructing a high-density linkage map and comparative maps with other legumes, and identifying quantitative trait loci (QTL) for levels of boron, calcium, iron, potassium, magnesium, manganese, molybdenum, phosphorous, sulfur, and zinc in the seed, as well as for seed weight. In this study, 1609 high quality SNPs were found to be polymorphic between 'Kiflica' and 'Aragorn', two parents of an F6-derived recombinant inbred line (RIL) population. Mapping 1683 markers including 75 previously published markers and 1608 SNPs developed from the present study generated a linkage map of size 1310.1 cM. Comparative mapping with other legumes demonstrated that the highest level of synteny was observed between pea and the genome of Medicago truncatula. QTL analysis of the RIL population across two locations revealed at least one QTL for each of the mineral nutrient traits. In total, 46 seed mineral concentration QTLs, 37 seed mineral content QTLs, and 6 seed weight QTLs were discovered. The QTLs explained from 2.4% to 43.3% of the phenotypic variance. The genome-wide SNPs and the genetic linkage map developed in this study permitted QTL identification for pea seed mineral

  15. Fatty acid profiling of the seed oils of some varieties of field peas (Pisum sativum) by RP-LC/ESI-MS/MS: towards the development of an oilseed pea.

    PubMed

    Villalobos Solis, Manuel Ivan; Patel, Anil; Orsat, Valérie; Singh, Jaswinder; Lefsrud, Mark

    2013-08-15

    Reversed-phase liquid chromatography coupled to negative-ion electrospray tandem mass spectrometry (RP-LC/ESI-MS/MS) was used to study the fatty acid profile from the oil of harvested field pea (Pisum sativum) varieties as part of a research project to develop this legume as a commercial oilseed for Canada. The seed oils from pea samples contained palmitic and stearic acids as major saturated fatty acids. Oleic, linoleic and linolenic acids were the major unsaturated fatty acids found. Small percentages of other long chain fatty acids were also detected. This profile suggests that the species of field pea investigated might have the potential to be used as raw materials to develop a future new oilseed crop for the food industry. Fatty acid extracts did not require further manipulation before final analysis by RP-LC/ESI-MS/MS, indicating the utility and relative simplicity of this technique for future screening studies. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Stomatal response and leaf injury of Pisum sativum L. with SO/sub 2/ and O/sub 3/ exposures. II. Influence of moisture stress and time of exposure

    SciTech Connect

    Olszyk, D.M.; Tibbitts, T.W.

    1981-03-01

    Stomatal response during exposure to SO/sub 2/ and O/sub 3/ and subsequent leaf injury were examined in plants of Pisum sativum L. Alsweet grown in a peat-vermiculite medium in controlled environment chambers. Plants developing under moisture stress, induced by drying the medium to 50% of field capacity, exhibited greater stomatal closure during exposures and less than one-fourth the necrosis compared to plants developing in a medium maintained at field capacity. Plants under moisture stress had only a slightly more negative plant water potential (approx. = 4.0 bars) than at field capacity (approx. = 3.4 bars). Plants exposed to pollutants for 2 hours near the beginning or end of a 16-hour light period had greater stomatal closure during exposures and less leaf necrosis than plants exposed during the middle of the light period.

  17. cDNA cloning, primary structure and gene expression for H-protein, a component of the glycine-cleavage system (glycine decarboxylase) of pea (Pisum sativum) leaf mitochondria.

    PubMed Central

    Macherel, D; Lebrun, M; Gagnon, J; Neuburger, M; Douce, R

    1990-01-01

    We have isolated and characterized cDNA clones encoding the H-protein of the glycine-cleavage system of pea (Pisum sativum) leaf mitochondria. The deduced primary structure revealed that the 131-amino-acid polypeptide is cytoplasmically synthesized with a 34-amino-acid mitochondrial targeting peptide. The lipoate-binding site was assigned to be lysine-63, as deduced from a sequence comparison with several lipoate-bearing proteins. The expression of the gene encoding H-protein was shown to occur specifically in the leaf tissue, with light exerting an additional effect by increasing the mRNA levels severalfold. Two polyadenylation sites were found in the mRNA, and a single-copy gene encoding the H-protein was detected in pea genome. Images Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. PMID:2363710

  18. The isoperoxidases of Pisum sativum.

    PubMed

    Siegel, B Z; Galston, A W

    1967-02-01

    The heterogeneity of the peroxidases in peas was examined by starch gel electrophoresis. Comparisons were made between tall and dwarf cultivars and among organ systems developed in light and darkness. Isoperoxidase bands could be grouped as cathodic, anodic and near-neutral (at pH 9.0) types. The cathodic set stained well with guaiacol oxidation products whereas some anodic bands reacted preferentially with 2,6-dimethoxyphenol. Some near-neutral bands were aceto-carmine positive and may have been organellar. Each organ had a characteristic isozyme pattern, and the band patterns in corresponding organs from different varieties were far more alike than were the patterns in the different organs within each variety. Ontogenetic changes were marked in all 3 organ systems, principally in the cathodic bands. The effect of light on isozymal patterns was quantitative rather than qualitative, possibly influencing the isoperoxidases secondarily via its effect upon organ physiology and development.

  19. The Isoperoxidases of Pisum sativum

    PubMed Central

    Siegel, B. Z.; Galston, A. W.

    1967-01-01

    The heterogeneity of the peroxidases in peas was examined by starch gel electrophoresis. Comparisons were made between tall and dwarf cultivars and among organ systems developed in light and darkness. Isoperoxidase bands could be grouped as cathodic, anodic and near-neutral (at pH 9.0) types. The cathodic set stained well with guaiacol oxidation products whereas some anodic bands reacted preferentially with 2,6-dimethoxyphenol. Some near-neutral bands were aceto-carmine positive and may have been organellar. Each organ had a characteristic isozyme pattern, and the band patterns in corresponding organs from different varieties were far more alike than were the patterns in the different organs within each variety. Ontogenetic changes were marked in all 3 organ systems, principally in the cathodic bands. The effect of light on isozymal patterns was quantitative rather than qualitative, possibly influencing the isoperoxidases secondarily via its effect upon organ physiology and development. PMID:6040893

  20. Accumulation of Phosphorus-Containing Compounds in Developing Seeds of Low-Phytate Pea (Pisum sativum L.) Mutants

    PubMed Central

    Shunmugam, Arun S.K.; Bock, Cheryl; Arganosa, Gene C.; Georges, Fawzy; Gray, Gordon R.; Warkentin, Thomas D.

    2014-01-01

    Low phytic acid (lpa) crops are low in phytic acid and high in inorganic phosphorus (Pi). In this study, two lpa pea genotypes, 1-150-81, 1-2347-144, and their progenitor CDC Bronco were grown in field trials for two years. The lpa genotypes were lower in IP6 and higher in Pi when compared to CDC Bronco. The total P concentration was similar in lpa genotypes and CDC Bronco throughout the seed development. The action of myo-inositol phosphate synthase (MIPS) (EC 5.5.1.4) is the first and rate-limiting step in the phytic acid biosynthesis pathway. Aiming at understanding the genetic basis of the lpa mutation in the pea, a 1530 bp open reading frame of MIPS was amplified from CDC Bronco and the lpa genotypes. Sequencing results showed no difference in coding sequence in MIPS between CDC Bronco and lpa genotypes. Transcription levels of MIPS were relatively lower at 49 days after flowering (DAF) than at 14 DAF for CDC Bronco and lpa lines. This study elucidated the rate and accumulation of phosphorus compounds in lpa genotypes. The data also demonstrated that mutation in MIPS was not responsible for the lpa trait in these pea lines. PMID:27135314

  1. Osmotic adjustment and the growth response of seven vegetable crops following water-deficit stress. [Phaseolus vulgaris L. ; Beta vulgaris L. ; Abelmoschus esculentus; Pisum sativum L. ; Capsicum annuum L. ; Spinacia oleracea L. ; Lycopersicon esculentum Mill

    SciTech Connect

    Wullschleger, S.D. ); Oosterhuis, D.M. )

    1991-09-01

    Growth-chamber studies were conducted to examine the ability of seven vegetable crops- Blue Lake beam (Phaseolus vulgaris L.) Detroit Dark Red beet (Beta vulgaris L.) Burgundy okra (Abelmoschus esculentus) (Moench), Little Marvel pea (Pisum sativum L), California Wonder bell pepper (Capsicum annuum L), New Zealand spinach (Spinacia oleracea L), and Beefsteak tomato (Lycopersicon esculentum Mill.) - to adjust osmotically in response to water-deficit stress. Water stress was imposed by withholding water for 3 days, and the adjustment of leaf and root osmotic potentials upon relief of the stress and rehydration were monitored with thermocouple psychrometers. Despite similar reductions in leaf water potential and stomatal conductance among the species studied reductions in lead water potential an stomatal conductance among the species, crop-specific differences were observed in leak and root osmotic adjustment. Leaf osmotic adjustment was observed for bean, pepper, and tomato following water-deficit stress. Root osmotic adjustment was significant in bean, okra, pea and tomato. Furthermore, differences in leaf and root osmotic adjustment were also observed among five tomato cultivars. Leaf osmotic adjustment was not associated with the maintenance of leaf growth following water-deficit stress, since leaf expansion of water-stressed bean and pepper, two species capable of osmotic adjustment, was similar to that of spinach, which exhibited no leaf osmotic adjustment.

  2. Phylogenetic diversity based on rrs, atpD, recA genes and 16S-23S intergenic sequence analyses of rhizobial strains isolated from Vicia faba and Pisum sativum in Peru.

    PubMed

    Santillana, Nery; Ramírez-Bahena, Martha Helena; García-Fraile, Paula; Velázquez, Encarna; Zúñiga, Doris

    2008-03-01

    In this study 17 isolates from effective nodules of Vicia faba and Pisum sativum var. macrocarpum growing in different soils from Peru were isolated and characterized. The isolates, presenting 11 different RAPD profiles, were distributed in three groups on the basis of their 16S-RFLP patterns. The 16S rRNA gene sequences of strains from 16S-RFLP groups I, II and III were closely related (identities higher than 99.5%) to Rhizobium leguminosarum bv. trifolii DSM 30141 (=ATCC 14480), R. leguminosarum bv. viciae DSM 30132(T) and Rhizobium etli CFN42(T) (=USDA 9032(T)), respectively. The analysis of the 16S-23S intergenic spacer (ITS) and two housekeeping genes, atpD and recA, confirmed the identification of strains from group I, however those from groups II and III were phylogenetically divergent to strains DSM 30132(T) and CFN42(T). These results support the fact that the 16S rRNA gene is not adequate for identification at species level within genus Rhizobium and suggest the existence of putative new species within the phylogenetic group of R. leguminosarum. They also confirm the need of a taxonomic revision of R. leguminosarum since the reference strains of the three biovars included in this study are phylogenetically divergent according to their ITS, atpD and recA gene sequences.

  3. Peptides from Pisum sativum L. enzymatic protein digest with anti-adhesive activity against Helicobacter pylori: structure-activity and inhibitory activity against BabA, SabA, HpaA and a fibronectin-binding adhesin.

    PubMed

    Niehues, Michael; Euler, Marco; Georgi, Gilda; Mank, Marko; Stahl, Bernd; Hensel, Andreas

    2010-12-01

    Identification of anti-adhesive peptides against Helicobacter pylori obtained by enzymatic hydrolysis of seed proteins from Pisum sativum L. (Fabaceae). Bioassay-guided fractionation of protein tryptic digest by ultrafiltration, size exclusion chromatography (SEC) and reversed phase chromatography (RPC) were used. Identification of bioactive peptides was achieved by MALDI-TOF-MS. Adhesion of H. pylori was monitored by two different assays, using a quantitative in vitro assay on human AGS cells with evaluation of bacterial binding by flow cytometry, beside a semi-quantitative in situ adhesion assay using FITC-labelled H. pylori on human stomach tissue sections. From two highly active fractions (F3, F3.3) two anti-adhesive peptides (S3, S5) were identified. Neither F3 nor S3 or S5 had any cytotoxic effect against H. pylori. By hemagglutination assay and semiquantitative dot blot overlay assay with immobilized ligands it was shown that F3 interacts specifically with H. pylori adhesins BabA, SabA, HpaA and a fibronectin-binding adhesin, while S3 and S5 inhibit only BabA. It was demonstrated that BabA, usually interacting with carbohydrate motifs such as fucosylated blood group antigens, interacts with the peptide moieties. Bioactive peptides from pea protein could be applied as functional ingredients for protecting infants and children against infections such as H. pylori. Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Response to water deficit and high temperature of transgenic peas (Pisum sativum L.) containing a seed-specific alpha-amylase inhibitor and the subsequent effects on pea weevil (Bruchus pisorum L.) survival.

    PubMed

    Sousa-Majer, Maria José de; Turner, Neil C; Hardie, Darryl C; Morton, Roger L; Lamont, Byron; Higgins, Thomas J V

    2004-02-01

    The effects of water deficit and high temperature on the production of alpha-amylase inhibitor 1 (alpha-AI-1) were studied in transgenic peas (Pisum sativum L.) that were developed to control the seed-feeding pea weevil (Bruchus pisorum L., Coleoptera: Bruchidae). Transgenic and non-transgenic plants were subjected to water-deficit and high-temperature treatments under controlled conditions in the glasshouse and growth cabinet, beginning 1 week after the first pods were formed. In the water-deficit treatments, the peas were either adequately watered (control) or water was withheld after first pod formation. The high-temperature experiments were performed in two growth cabinets, one maintained at 27/22 degrees C (control) and one at 32/27 degrees C day/night temperatures, with the vapour pressure deficit maintained at 1.3 kPa. The plants exposure to high temperatures and water deficit produced 27% and 79% fewer seeds, respectively, than the controls. In the transgenic peas the level of alpha-AI-1 as a percentage of total protein was not influenced by water stress, but was reduced on average by 36.3% (the range in two experiments was 11-50%) in the high-temperature treatment. Transgenic and non-transgenic pods of plants grown at 27/22 degrees C and 32/27 degrees C were inoculated with pea weevil eggs to evaluate whether the reduction in level of alpha-AI-1 in the transgenic pea seeds affected pea weevil development and survival. At the higher temperatures, 39% of adult pea weevil emerged, compared to 1.2% in the transgenic peas grown at the lower temperatures, indicating that high temperature reduced the protective capacity of the transgenic peas.

  5. Genetic diversity in European Pisum germplasm collections.

    PubMed

    Jing, R; Ambrose, M A; Knox, M R; Smykal, P; Hybl, M; Ramos, Á; Caminero, C; Burstin, J; Duc, G; van Soest, L J M; Święcicki, W K; Pereira, M G; Vishnyakova, M; Davenport, G F; Flavell, A J; Ellis, T H N

    2012-07-01

    The distinctness of, and overlap between, pea genotypes held in several Pisum germplasm collections has been used to determine their relatedness and to test previous ideas about the genetic diversity of Pisum. Our characterisation of genetic diversity among 4,538 Pisum accessions held in 7 European Genebanks has identified sources of novel genetic variation, and both reinforces and refines previous interpretations of the overall structure of genetic diversity in Pisum. Molecular marker analysis was based upon the presence/absence of polymorphism of retrotransposon insertions scored by a high-throughput microarray and SSAP approaches. We conclude that the diversity of Pisum constitutes a broad continuum, with graded differentiation into sub-populations which display various degrees of distinctness. The most distinct genetic groups correspond to the named taxa while the cultivars and landraces of Pisum sativum can be divided into two broad types, one of which is strongly enriched for modern cultivars. The addition of germplasm sets from six European Genebanks, chosen to represent high diversity, to a single collection previously studied with these markers resulted in modest additions to the overall diversity observed, suggesting that the great majority of the total genetic diversity collected for the Pisum genus has now been described. Two interesting sources of novel genetic variation have been identified. Finally, we have proposed reference sets of core accessions with a range of sample sizes to represent Pisum diversity for the future study and exploitation by researchers and breeders.

  6. [Molecular genetic diversity of the pea (Pisum sativum L.) from the collection of N.I. Vavilov Research Institute of Plant Industry as detected by AFLP analysis].

    PubMed

    D'iachenko, E A; Ryzhova, N N; Vishniakova, M A; Kochieva, E Z

    2014-09-01

    In this work, a pea selection obtained from the Vavilov Research Institute collection comprising 83 P. sativum samples, including representatives of three subspecies, was studied using the ?FL? labeling method. Local cultivars for different uses with maximum ecological and geographical diversity (including those from the centers of origin of the species) were predominantly chosen for the study; a number of their morphological and biological characteristics were evaluated. We obtained 382 polymorphic AFLP fragments; each sample was characterized by a unique set of these fragments. The genetic diversity of the studied material was evaluated, and a wide range of genetic differences in the investigated samples (0.07-0.27) was demonstrated. The affiliation of the samples to the certain subspecies was not confirmed by the obtained data; the ecogeographical differentiation of the samples was not reflected by the data. Factor analysis allowed us to identify the sample groups of European and Asian origin and the intermediate nature of most of the samples from the studied selection.

  7. Cloning and functional characterization of the promoter of PsSEOF1 gene from Pisum sativum under different stress conditions using Agrobacterium-mediated transient assay.

    PubMed

    Srivastava, Vineet Kumar; Raikwar, Shailendra; Tuteja, Narendra

    2014-01-01

    PsSEOF1, a SEO (sieve element occlusion) gene family protein (forisome) is calcium powered motor protein and is located close to plasma membrane of sieve element. In sieve element (SE) it senses the calcium ion levels and undergoes ATP-independent conformational shifts. Forisome, meaning gate-bodies (Latin foris: wing of a gate; Greek soma: body). Recent reports show that SEO gene family protein can prevent the loss of nutrient rich photoassimilate upon wound injury. The regulation of SEO protein forisome under abiotic/ biotic stress is still unknown. The analysis of cis-regulatory element present in the upstream region is not well understood. Tissue specific promoters guarantee correct expression when it perceives particular stimuli. Here we report isolation of tissue specific promoter of PsSEOF1 was isolated by gene walking PCR from P. sativum (pea) genomic DNA library constructed by BD genome walker kit. In silico analysis revealed several putative cis element within this promoter sequence like wound response, cold, dehydration. Putative elements which might be required for its vascular tissue specificity has also been identified. The GUS activities of PsSEOF1 promoter-GUS chimeric construct in the agroinfiltrated leaves under different environmental stress abiotic and biotic like wound, cold, salt and phytohormones has shown high level of GUS activity. To identify the activity of PsSEOF1 promoter under different stress condition an Agrobacterium-mediated transient expression of tobacco plants were subjected to histochemical GUS staining. Stress-inducible nature of PsSEOF1 promoter opens possibility for the study of the PsSEOF1 gene regulation under stress condition. The isolated promoter sequence could serve as an important candidate for tissue specific promoter in genetic engineering of plant under stress conditions.

  8. Zoospore Homing and Infection Events: Effects of the Biocontrol Bacterium Burkholderia cepacia AMMDR1 on Two Oomycete Pathogens of Pea (Pisum sativum L.)

    PubMed Central

    Heungens, K.; Parke, J. L.

    2000-01-01

    Burkholderia cepacia AMMDR1 is a biocontrol agent that protects pea and sweet corn seeds from Pythium damping-off in field experiments. The goal of this work was to understand the effect of B. cepacia AMMDR1 on Pythium aphanidermatum and Aphanomyces euteiches zoospore homing events and on infection of pea seeds or roots. In vitro, B. cepacia AMMDR1 caused zoospore lysis, prevented cyst germination, and inhibited germ tube growth of both oomycetes. B. cepacia AMMDR1 also reduced the attractiveness of seed exudates to Pythium zoospores to nondetectable levels. However, when present at high levels on seeds, B. cepacia AMMDR1 had little net effect on zoospore attraction, probably because it also enhanced seed exudation. Seed-applied B. cepacia AMMDR1 dramatically reduced the incidence of infection by Pythium zoospores in situ compared with an antibiosis-deficient Tn5 mutant strain. This mutant strain also decreased Pythium infection incidence to some extent, but only when the pathogen inoculum potential was low. B. cepacia AMMDR1 did not affect attraction of Aphanomyces zoospores or Aphanomyces root rot incidence. These results suggest that B. cepacia AMMDR1 controls P. aphanidermatum largely through antibiosis, but competition for zoospore-attracting compounds can contribute to the effect. Differences in suppression of Aphanomyces and Pythium are discussed in relation to differences in the ecology of the two pathogens. PMID:11097889

  9. Biological safety assessment of mutant variant of Allium sativum leaf agglutinin (mASAL), a novel antifungal protein for future transgenic application.

    PubMed

    Ghosh, Prithwi; Roy, Amit; Chakraborty, Joydeep; Das, Sampa

    2013-12-04

    Genetic engineering has established itself to be an important tool for crop improvement. Despite the success, there is always a risk of food allergy induced by alien gene products. The present study assessed the biosafety of mutant Allium sativum leaf agglutinin (mASAL), a potent antifungal protein generated by site directed mutagenesis of Allium sativum leaf agglutinin (ASAL). mASAL was cloned in pET28a+ and expressed in E. coli, and the safety assessment was carried out according to the FAO/WHO guideline (2001). Bioinformatics analysis, pepsin digestion, and thermal stability assay showed the protein to be nonallergenic. Targeted sera screening revealed no significant IgE affinity of mASAL. Furthermore, mASAL sensitized Balb/c mice showed normal histopathology of lung and gut tissue. All results indicated the least possibility of mASAL being an allergen. Thus, mASAL appears to be a promising antifungal candidate protein suitable for agronomical biotechnology.

  10. Biochemical responses to Fe deficiency in Pisum sativum L., cv. Sparkle and dgl mutant: Is the PM H+-ATPase the sole activity involved in H+ extrusion?

    USDA-ARS?s Scientific Manuscript database

    The study of processes regulating Fe acquisition by plants provides useful knowledge for breeding programs aimed to obtain Fe-efficient and/or biofortified varieties. In fact, total Fe uptake is an important, though not sufficient prerequisite to increase Fe density in plant tissues. Like the majori...

  11. Ferric reductase activity and PsFRO1 sequence variation in pisum sps

    USDA-ARS?s Scientific Manuscript database

    Physiological studies in pea (Pisum sativum) suggest that the reduction of iron (Fe) is the rate-limiting physiological process in Fe acquisition by dicotyledonous plants. Previous molecular work suggests that ferric reductase activity is regulated at both the transcriptional and post-translational ...

  12. Cytokinins in Pisum Transfer Ribonucleic Acid 1

    PubMed Central

    Vreman, Hendrik J.; Skoog, Folke; Frihart, Charles R.; Leonard, Nelson J.

    1972-01-01

    Five cytokinin-active ribonucleosides have been isolated from the transfer RNA of 7-day-old green pea shoots (Pisum sativum L. var. Alaska). Ultraviolet spectroscopy and mass spectrometry have been used to identify 6-(3-methyl-2-butenylamino)-9-β-d-ribofuranosylpurine, 6-(4-hydroxy-3-methyl-2-butenylamino)-2-methylthio-9-β- d-ribofuranosylpurine, and 6-(4-hydroxy-3-methyl-2-butenylamino)-9-β-d-ribofuranosylpurine. The latter was separated into the cis- and trans-isomers by thin layer chromatography. The fifth cytokinin is indicated to be 6-(3-methyl-2-butenylamino)-2-methylthio-9-β-d -ribofuranosylpurine on the basis of its chromatographic properties. PMID:16658059

  13. Genetic Variation Controlling Wrinkled Seed Phenotypes in Pisum: How Lucky Was Mendel?

    PubMed

    Rayner, Tracey; Moreau, Carol; Ambrose, Mike; Isaac, Peter G; Ellis, Noel; Domoney, Claire

    2017-06-06

    One of the traits studied by Mendel in pea (Pisum sativum L.) was the wrinkled-seeded phenotype, and the molecular basis for a mutation underlying this phenotype was discovered in the 1990s. Although the starch-branching enzyme gene mutation identified at the genetic locus r is most likely to be that in seeds available to Mendel in the mid-1800s, it has remained an open question as to whether or not additional natural mutations in this gene exist within Pisum germplasm collections. Here, we explore this question and show that all but two wrinkled-seeded variants in one such collection correspond to either the mutant allele described previously for the r locus or a mutation at a second genetic locus, rb, affecting the gene encoding the large subunit of Adenosine diphosphoglucose (ADP-glucose) pyrophosphorylase; the molecular basis for the rb mutation is described here. The genetic basis for the phenotype of one (JI 2110) of the two lines which are neither r nor rb has been studied in crosses with a round-seeded variant (JI 281); for which extensive genetic marker data were expected. In marked contrast to the trait studied by Mendel and the rb phenotype; the data suggest that the wrinkled-seeded phenotype in JI 2110 is maternally determined, controlled by two genetic loci, and the extent to which it is manifested is very sensitive to the environment. Metabolite analysis of the cotyledons of JI 2110 revealed a profile for sucrose and sucrose-derived compounds that was more similar to that of wild-type round-seeded, than that of wrinkled-seeded r, pea lines. However, the metabolite profile of the seed coat (testa) of JI 2110 was distinct from that of other round-seeded genotypes tested which, together with analysis of recombinant inbred progeny lines, suggests an explanation for the seed phenotype.

  14. The genetic diversity and evolution of field pea (Pisum) studied by high throughput retrotransposon based insertion polymorphism (RBIP) marker analysis

    PubMed Central

    2010-01-01

    pools into which the genus Pisum is partitioned and their geographical distribution. The data strongly support the model of independent domestications for P. sativum ssp abyssinicum and P. sativum. The relationships between these two cultivated germplasms and the various sub-divisions of wild Pisum have been clarified and the most likely ancestral wild gene pools for domesticated P. sativum identified. Lastly, this study provides a framework for defining global Pisum germplasm which will be useful for designing core collections. PMID:20156342

  15. Micromonospora luteifusca sp. nov. isolated from cultivated Pisum sativum.

    PubMed

    Carro, Lorena; Riesco, Raúl; Spröer, Cathrin; Trujillo, Martha E

    2016-06-01

    Three novel actinobacterial strains, GUI2(T), GUI42 and CR21 isolated from nodular tissues and the rhizosphere of a sweet pea plant collected in Cañizal, Spain were identified according to their 16S rRNA gene sequences as new members of the genus Micromonospora. The closest phylogenetic members were found to be Micromonospora saelicesensis (99.2%) "Micromonospora zeae" (99.1%), "Micromonospora jinlongensis" (99%), Micromonospora lupini (98.9%) and Micromonospora zamorensis (98.8%). To resolve their full taxonomic position, four additional genes (atpD, gyrB, recA, rpoB) were partially sequenced and compared to available Micromonospora type strain sequences. DNA-DNA hybridization, BOX-PCR and ARDRA profiles confirmed that these strains represent a novel genomic species. All strains contained meso-diaminopimelic and hydroxy-diaminopimelic acids in their cell wall. Their fatty acid profiles comprised iso-C15:0, iso-C16:0 and anteiso-C15:0 as major components. The polar lipids diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol were found in the type strain GUI2(T) which also contained MK-10(H4) as the major menaquinone. Physiological and biochemical characteristics also differentiated the new isolates. Based on the integration of the above studies, strains GUI2(T), GUI42 and CR21 represent a novel Micromonospora species and we propose the name Micromonospora luteifusca sp. nov. The type strain is GUI2(T) (=CECT 8846(T); =DSM 100204(T)).

  16. Ethylene binding in epicotyls of Pisum sativum L. cv. Alaska.

    PubMed

    Sanders, I O; Smith, A R; Hall, M A

    1991-01-01

    Using in-vivo assays at least two classes of ethylene-binding site are shown to exist in pea epicotyls. These classes appear to have identical affinities for ethylene with a KD of 6 · 10(-11)-8 · 10(-11) M; the fast associating class has high and appropriate affinities for physiologically active analogues of ethylene, and CO2 is without effect upon bfinding. Experiments involving suppression of endogenous ethylene production demonstrate that previous work may have underestimated both the amount of binding sites present and the affinity of such sites for ethylene. The apparent inhibition of binding by silver appears to be the consequence of a stimulation of endogenous ethylene production. The possible role of the sites as ethylene receptors is discussed.

  17. Whole shoot mineral partitioning and accumulation in pea (Pisum sativum)

    USDA-ARS?s Scientific Manuscript database

    Several grain legumes are staple food crops that are important sources of minerals for humans; unfortunately, our knowledge is incomplete with respect to the mechanisms of translocation of these minerals to the vegetative tissues and loading into seeds. Understanding the mechanism and partitioning o...

  18. Excision and replication of extrachromosomal DNA of pea (Pisum sativum)

    SciTech Connect

    Hof, J.V.; Bjerknes, C.A.; Delihas, N.C.

    1983-02-01

    Experiments with cultured pea roots were conducted to determine (i) whether extrachromosomal DNA was produced by cells in the late S phase or in the G2 phase of the cell cycle, (ii) whether the maturation of nascent DNA replicated by these cells achieved chromosomal size, (iii) when extrachromosomal DNA was removed from the chromosomal duplex, and (iv) the replication of nascent chains by the extrachromosomal DNA after its release from the chromosomal duplex. Autoradiography and cytophotometry of cells of carbohydrate-starved root tips revealed that extrachromosomal DNA was produced by a small fraction of cells accumulated in the late S phase after they had replicated about 80% of their DNA. Velocity sedimentation of nascent chromosomal DNA in alkaline sucrose gradients indicated that the DNA of cells in the late S phase failed to achieve chromosomal size. After reaching sizes of 70 x 10/sup 6/ to 140 x 10/sup 6/ daltons, some of the nascent chromosomal molecules were broken, presumably releasing extrachromosomal DNA several hours later. Sedimentation of selectively extracted extrachromosomal DNA either from dividing cells or from those in the late S phase showed that it replicated two nascent chains, one of 3 x 10/sup 6/ daltons and another of 7 x 10/sup 6/ daltons. Larger molecules of extrachromosomal DNA were detectable after cells were labeled for 24 h. These two observations were compatible with the idea that the extrachromosomal DNA was first replicated as an integral part of the chromosomal duplex, was cut from the duplex, and then, once free of the chromosome, replicated two smaller chains of 3 x 10/sup 6/ and 7 x 10/sup 6/ daltons. 24 references.

  19. Physiological effects and uptake of cadmium in Pisum sativum

    SciTech Connect

    Wong, Y.S.; Lam, H.M.; Dhillon, E.; Tam, N.F.Y.; Leung, W.N.

    1988-01-01

    Germination and elongation of pea radicles were significantly inhibited when Cd was added to the culture solutions. The heavy metal also retarded shoot elongation and leaf development of young seedlings even at a concentration of 1 {mu}g/mL. In addition to chlorosis and leaf wilting, Cd caused severe stem constriction. This observation together with the ultrastructural studies on the vascular system indicated that Cd damaged growth narrowing of vessel pits and deposition of unknown debris which blocked the water translocation flow. The degree of inhibition on root and shoot growth normally increased with an increase in Cd concentration contained in the culture medium, although mild enhancement of growth was also recorded when Cd was added at a level of 25 ng/L. In all parts of the plants, the amount of Cd uptaken was directly related to the external level of Cd. Despite 70 to 80% of the total Cd accumulated was confined to the root portion, the stem and the leaf appeared to be more severely affected than the root. This may suggest that the root is the most tolerant part which plays a role to sequester Cd from being transported to the upper parts of the plant.

  20. Whole shoot mineral partitioning and accumulation in pea (Pisum sativum).

    PubMed

    Sankaran, Renuka P; Grusak, Michael A

    2014-01-01

    Several grain legumes are staple food crops that are important sources of minerals for humans; unfortunately, our knowledge is incomplete with respect to the mechanisms of translocation of these minerals to the vegetative tissues and loading into seeds. Understanding the mechanism and partitioning of minerals in pea could help in developing cultivars with high mineral density. A mineral partitioning study was conducted in pea to assess whole-plant growth and mineral content and the potential source-sink remobilization of different minerals, especially during seed development. Shoot and root mineral content increased for all the minerals, although tissue-specific partitioning differed between the minerals. Net remobilization was observed for P, S, Cu, and Fe from both the vegetative tissues and pod wall, but the amounts remobilized were much below the total accumulation in the seeds. Within the mature pod, more minerals were partitioned to the seed fraction (>75%) at maturity than to the pod wall for all the minerals except Ca, where only 21% was partitioned to the seed fraction. Although there was evidence for net remobilization of some minerals from different tissues into seeds, continued uptake and translocation of minerals to source tissues during seed fill is as important, if not more important, than remobilization of previously stored minerals.

  1. Whole shoot mineral partitioning and accumulation in pea (Pisum sativum)

    PubMed Central

    Sankaran, Renuka P.; Grusak, Michael A.

    2014-01-01

    Several grain legumes are staple food crops that are important sources of minerals for humans; unfortunately, our knowledge is incomplete with respect to the mechanisms of translocation of these minerals to the vegetative tissues and loading into seeds. Understanding the mechanism and partitioning of minerals in pea could help in developing cultivars with high mineral density. A mineral partitioning study was conducted in pea to assess whole-plant growth and mineral content and the potential source-sink remobilization of different minerals, especially during seed development. Shoot and root mineral content increased for all the minerals, although tissue-specific partitioning differed between the minerals. Net remobilization was observed for P, S, Cu, and Fe from both the vegetative tissues and pod wall, but the amounts remobilized were much below the total accumulation in the seeds. Within the mature pod, more minerals were partitioned to the seed fraction (>75%) at maturity than to the pod wall for all the minerals except Ca, where only 21% was partitioned to the seed fraction. Although there was evidence for net remobilization of some minerals from different tissues into seeds, continued uptake and translocation of minerals to source tissues during seed fill is as important, if not more important, than remobilization of previously stored minerals. PMID:24795736

  2. Sucrose transport into developing seeds of Pisum sativum L.

    PubMed

    Tegeder, M; Wang, X D; Frommer, W B; Offler, C E; Patrick, J W

    1999-04-01

    The anatomy of developing pea seeds is characterized by transfer cells present in both coats and cotyledons at the maternal/filial interface. To determine the nature and cellular localization of sucrose transporters in pea seeds, a full-length clone of a sucrose/H+ symporter (PsSUT1) was isolated from a cotyledon cDNA library. Northern blot analyses of different organs showed that PsSUT1 is expressed in non-seed tissues, including sucrose sinks and sources. Within developing seeds, transcripts of PsSUT1 and PsAHA1 genes were detected in all tissues, while transcripts of a sucrose binding protein (GmSBP) were confined to cotyledon epidermal transfer cells. Signal intensities of PsSUT1 and PsAHA1 transcripts and protein products were most pronounced in the thin-walled parenchyma cells of seed coats and epidermal transfer cells of cotyledons. For cotyledons, the highest transporter densities were localized to those portions of plasma membranes lining the wall ingrowth regions of epidermal transfer cells. Responses of [14C]sucrose influx to metabolic inhibitors indicated that proton-coupled sucrose transport was operative in both seed coats and cotyledons. Cotyledon epidermal transfer cells were shown to support the highest sucrose flux. Maximal transport activity was found to account for the sucrose flux differences between seed tissues. Intercellular movement of the symplasmic tracer, 5-(6)-carboxyfluorescein (CF), demonstrated that symplasmic pathways interconnect the vascular tissues to thin-walled parenchyma transfer cells of seed coats and, for cotyledons, epidermal transfer cells to storage parenchyma cells.

  3. Brassinosteroid deficiency due to truncated steroid 5alpha-reductase causes dwarfism in the lk mutant of pea.

    PubMed

    Nomura, Takahito; Jager, Corinne E; Kitasaka, Yukiko; Takeuchi, Keiichi; Fukami, Motohiro; Yoneyama, Koichi; Matsushita, Yasuhiko; Nyunoya, Hiroshi; Takatsuto, Suguru; Fujioka, Shozo; Smith, Jennifer J; Kerckhoffs, L Huub J; Reid, James B; Yokota, Takao

    2004-08-01

    The endogenous brassinosteroids in the dwarf mutant lk of pea (Pisum sativum) were quantified by gas chromatography-selected ion monitoring. The levels of castasterone, 6-deoxocastasterone, and 6-deoxotyphasterol in lk shoots were reduced 4-, 70-, and 6-fold, respectively, compared with those of the wild type. The fact that the application of brassinolide restored the growth of the mutant indicated that the dwarf mutant lk is brassinosteroid deficient. Gas chromatography-selected ion monitoring analysis of the endogenous sterols in lk shoots revealed that the levels of campestanol and sitostanol were reduced 160- and 10-fold, respectively, compared with those of wild-type plants. These data, along with metabolic studies, showed that the lk mutant has a defect in the conversion of campest-4-en-3-one to 5alpha-campestan-3-one, which is a key hydrogenation step in the synthesis of campestanol from campesterol. This defect is the same as that found in the Arabidopsis det2 mutant and the Ipomoea nil kbt mutant. The pea gene homologous to the DET2 gene, PsDET2, was cloned, and it was found that the lk mutation would result in a putative truncated PsDET2 protein. Thus it was concluded that the short stature of the lk mutant is due to a defect in the steroidal 5alpha-reductase gene. This defect was also observed in the callus induced from the lk mutant. Biosynthetic pathways involved in the conversion of campesterol to campestanol are discussed in detail.

  4. Chromosome aberration assays in Pisum for the study of environmental mutagens.

    PubMed

    Grant, W F; Owens, E T

    2001-05-01

    From a literature survey, 117 chemicals are tabulated that have been assayed in 179 assays for their clastogenic effects in Pisum. Of the 117 chemicals that have been assayed, 65 are reported at giving a positive reaction (i.e. causing chromosome aberrations), 30 positive with a dose response, five borderline positive. Seventeen chemicals gave a negative response. Eighty-one percent of the chemicals gave a definite positive response. A c-mitotic effect was detected from treatment with 17 chemicals. In addition to the above tabulation of chemicals, 39 chemicals have been reported with an antimitotic effect. Thirteen assays have been recorded for five types of radiation, which with the exception of ultrasound reacted positively. The results of assays with 38 chemicals and/or radiations in combined treatments, as well as 15 chemicals and three types of radiations that induce somatic mutations are tabulated. The Pisum sativum (2n=14) bioassay has been shown to be a very good plant bioassay for assessing chromosome damage both in mitosis and meiosis for somatic mutations induced by chemicals, radiations, and environmental pollutants. For some chemicals, the Pisum assay is not as sensitive in assessing clastogenicity as the Allium assay, although this should be considered in relative terms. Pisum fulvum (2n=14) has been used in clastogenic studies also, but to a much lesser extent.

  5. The Temperature Response and Aggressiveness of Peyronellaea pinodes Isolates Originating from Wild and Domesticated Pisum sp. in Israel.

    PubMed

    Golani, M; Abbo, S; Sherman, A; Frenkel, O; Shtienberg, D

    2016-08-01

    Domesticated pea fields are grown in relatively close proximity to wild pea species in Israel. Despite the major role attributed to ascochyta blight in causing yield losses in domesticated pea, very limited information is available on the pathogens prevailing in natural ecosystems. The objectives of this study were (i) to identify the species causing ascochyta blight symptoms on leaves, stems, and petioles of domesticated pea and wild Pisum plants in Israel, and (ii) to quantify the temperature response(s) and aggressiveness of such pathogens originating from Pisum plants growing in sympatric and allopatric contexts. Eighteen fungal isolates were examined and identified; three of them were sampled from Pisum sativum, 11 from Pisum fulvum, and four from Pisum elatius. All isolates were identified as Peyronellaea pinodes. Spore germination and mycelial growth took place over a wide range of temperatures, the lower and upper cardinal temperatures being 2 to 9 and 33 to 38°C, respectively; the optimal temperatures ranged from 22 to 26°C. At an optimal temperature, disease severity was significantly higher for plants maintained under moist conditions for 24 h postinoculation than for those exposed to humidity for 5 or 10 h. Analyses of the data revealed that temperature responses, spore germination rates, and aggressiveness of isolates sampled from domesticated pea plants did not differ from those of isolates sampled from adjacent or distant wild populations. Host specificity was not observed. These observations suggest that Israel may be inhabited by a single metapopulation of P. pinodes.

  6. The dark-adaptation response of the de-etiolated pea mutant lip1 is modulated by external signals and endogenous programs.

    PubMed Central

    Frances, S; Thompson, W F

    1997-01-01

    The lip1 mutant of pea (Pisum sativum L.) exhibits a de-etiolated phenotype. When grown in darkness, lip1 plants have several characteristics normally associated only with light-grown plants. Young wild-type (WT) seedlings accumulate high levels of transcripts from plastid-related genes (such as those encoding chlorophyll a/b-binding proteins, ferredoxin, and the small subunit of Rubisco) only in the light. In contrast, regardless of the light conditions under which the plants are grown, young mutant seedlings accumulate transcript levels equal to or greater than those seen in light-grown WT seedlings of the same age. Under some conditions, light-grown lip1 seedlings failed to respond to dark treatment. The largest response to darkness observed in the mutant occurred when older seedlings were first grown under low-light conditions before transfer to darkness. The mutant's inability to respond to darkness is not due to a gross disturbance in the circadian clock. We conclude that environmental signals (light) and endogenous programs (developmental and circadian) regulate gene expression in both WT and mutant plants. However, mutant seedlings exhibit a developmentally regulated and exaggerated response to light. In addition, the effect of the mutation may be greatest during a brief period early in development. PMID:9306689

  7. Conditional Facilitation of an Aphid Vector, Acyrthosiphon pisum, by the Plant Pathogen, Pea Enation Mosaic Virus

    PubMed Central

    Hodge, Simon; Powell, Glen

    2010-01-01

    Plant pathogens can induce symptoms that affect the performance of insect herbivores utilizing the same host plant. Previous studies examining the effects of infection of tic bean, Vicia faba L. (Fabales: Fabaceae), by pea enation mosaic virus (PEMV), an important disease of legume crops, indicated there were no changes in the growth and reproductive rate of its primary vector the pea aphid, Acyrthosiphon pisum (Harris) (Hemiptera: Aphididae). Here, we report the results of laboratory experiments investigating how A. pisum responded to PEMV infection of a different host plant, Pisum sativum L., at different stages of symptom development. Aphid growth rate was negatively related to the age of the host plant, but when they were introduced onto older plants with well-developed PEMV symptoms they exhibited a higher growth rate compared to those developing on uninfected plants of the same age. In choice tests using leaf discs A. pisum showed a strong preference for discs from PEMV-infected peas, probably in response to visual cues from the yellowed and mottled infected leaves. When adults were crowded onto leaves using clip-cages they produced more winged progeny on PEMV-infected plants. The results indicate that PEMV produces symptoms in the host plant that can enhance the performance of A. pisum as a vector, modify the production of winged progeny and affect their spatial distribution. The findings provide further evidence that some insect vector/plant pathogen interactions could be regarded as mutualistic rather than commensal when certain conditions regarding the age, stage of infection and species of host plant are met. PMID:21067425

  8. Oxidative stress in pea seedling leaves in response to Acyrthosiphon pisum infestation.

    PubMed

    Mai, Van Chung; Bednarski, Waldemar; Borowiak-Sobkowiak, Beata; Wilkaniec, Barbara; Samardakiewicz, Sławomir; Morkunas, Iwona

    2013-09-01

    In this study we examined whether and to what extent oxidative stress is induced in seedling leaves of Pisum sativum L. cv. Cysterski in response to pea aphid (Acyrthosiphon pisum Harris) infestation. A. pisum caused oxidative stress conditions in pea leaves through enhanced production of the reactive oxygen species (ROS) hydrogen peroxide (H2O2) and superoxide anion radical (O2(·-)). Early, strong generation of H2O2 was observed at 24h in aphid-infested leaves. The highest level of H2O2 at this time point may be related to the functioning of H2O2 as a signaling molecule, triggering defense mechanisms in pea leaves against A. pisum. Additionally, the strong generation and continuous increase of O2(·-) production in aphid-infested leaves from 0 to 96 h enhanced the defense potential to protect against aphid herbivory. Also in the study cytochemical localization of H2O2 and O2(·-) in pea leaves after aphid infestation was determined using the confocal microscope. Relative release of H2O2 and O2(·-) was estimated by staining leaves with specific fluorochromes, i.e. dichlorodihydro-fluorescein diacetate (DCFH-DA) and dihydroethidium (DHE), respectively. DCFH-DA and DHE derived fluorescence was observed to cover a much larger tissue area in aphid-infested leaves, whereas little or no fluorescence was observed in the control leaves. Enhanced activity of the antioxidant enzymes superoxide dismutase (SOD, 1.15.1.1) and catalase (CAT, 1.11.1.6) is one of the most essential elements of defense responses in pea seedling leaves to oxidative stress. Additionally, generation of semiquinones, stable free radicals with g-values of 2.0020 and 2.0035, detected by electron paramagnetic resonance spectroscopy (EPR), was suggested as a protective action of pea that may contribute to build-up of a defensive barrier or activate other defense mechanisms. Concentrations of semiquinone radicals in aphid-infested seedling leaves not only were generally higher than in the control plants

  9. Brassinosteroid Deficiency Due to Truncated Steroid 5α-Reductase Causes Dwarfism in the lk Mutant of Pea1

    PubMed Central

    Nomura, Takahito; Jager, Corinne E.; Kitasaka, Yukiko; Takeuchi, Keiichi; Fukami, Motohiro; Yoneyama, Koichi; Matsushita, Yasuhiko; Nyunoya, Hiroshi; Takatsuto, Suguru; Fujioka, Shozo; Smith, Jennifer J.; Kerckhoffs, L. Huub J.; Reid, James B.; Yokota, Takao

    2004-01-01

    The endogenous brassinosteroids in the dwarf mutant lk of pea (Pisum sativum) were quantified by gas chromatography-selected ion monitoring. The levels of castasterone, 6-deoxocastasterone, and 6-deoxotyphasterol in lk shoots were reduced 4-, 70-, and 6-fold, respectively, compared with those of the wild type. The fact that the application of brassinolide restored the growth of the mutant indicated that the dwarf mutant lk is brassinosteroid deficient. Gas chromatography-selected ion monitoring analysis of the endogenous sterols in lk shoots revealed that the levels of campestanol and sitostanol were reduced 160- and 10-fold, respectively, compared with those of wild-type plants. These data, along with metabolic studies, showed that the lk mutant has a defect in the conversion of campest-4-en-3-one to 5α-campestan-3-one, which is a key hydrogenation step in the synthesis of campestanol from campesterol. This defect is the same as that found in the Arabidopsis det2 mutant and the Ipomoea nil kbt mutant. The pea gene homologous to the DET2 gene, PsDET2, was cloned, and it was found that the lk mutation would result in a putative truncated PsDET2 protein. Thus it was concluded that the short stature of the lk mutant is due to a defect in the steroidal 5α-reductase gene. This defect was also observed in the callus induced from the lk mutant. Biosynthetic pathways involved in the conversion of campesterol to campestanol are discussed in detail. PMID:15286289

  10. Ethylene Inhibitors Partly Restore Nodulation to Pea Mutant E 107 (brz) 1

    PubMed Central

    Guinel, Frédérique C.; LaRue, Thomas A.

    1992-01-01

    E107 (brz) is a pleiotropic mutant of pea (Pisum sativum L. cv Sparkle) characterized by low nodulation, leaf necrosis, excessive ion accumulation, and decreased plant size. The defective nodulation of E107 was studied by light microscopy of lateral roots. The number of infections per centimeter of lateral root was only a third that of Sparkle. Moreover, most of the infections were aborted early; i.e. in only 14% of the infections did the infection thread penetrate beyond the epidermis. Nodulation of E107 was partly restored by treating the plant with the ethylene inhibitors aminoethoxyvinylglycine (AVG) or Ag+. Treatment with Ag+ did not increase the number of infections, but half of the infections went to completion. Ag+ and AVG did not alter the size of the mutant, the accumulation of cations in its shoots, nor the leaf necrosis. Thus, in E107, nodule development can be uncoupled from other pleiotropic characteristics. ImagesFigure 1Figure 2Figure 3 PMID:16668916

  11. Mutant alleles at the rugosus loci in pea affect seed moisture sorption isotherms and the relations between seed longevity and moisture content.

    PubMed

    Lyall, T W; Ellis, R H; John, P; Hedley, C L; Wang, T L

    2003-01-01

    Pea (Pisum sativum L.) mutant near-isogenic lines (RRrbrb, rrRbRb, rrrbrb) with lower starch but higher lipid contents, brought about by lesions in the starch biosynthetic pathway, had seed moisture sorption isotherms displaced below that of the wild type (RRRbRb). The negative logarithmic relationship between seed longevity and seed storage moisture content (%, f.wt basis), determined in hermetic storage at 65 degrees C, also differed: longevity in the mutant near-isogenic lines was poorer and less sensitive to moisture content than in the wild type (i.e. C(W) was lower). The low-moisture-content limit (m(c)) to this relation also differed, being lower in the mutant near-isogenic lines (5.4-5.9%) than in the wild type (6.1%). In contrast, all four near-isogenic lines showed no difference (P >0.25) in the negative semi-logarithmic relationship between equilibrium relative humidity (ERH) and seed longevity. It is concluded that the effect of these alleles at the r and rb loci on seed longevity was largely indirect; a consequence of their effect on seed composition and hence on moisture sorption isotherms. However, this explanation could not be invoked at moisture contents below m(c) where differences in longevity remained substantial (RRRbRb double that of rrrbrb). Hence, these mutant alleles affected seed longevity directly at very low moisture contents.

  12. EMS mutagenesis in the pea aphid Acyrthosiphon pisum.

    PubMed

    Tagu, Denis; Le Trionnaire, Gaël; Tanguy, Sylvie; Gauthier, Jean-Pierre; Huynh, Jean-René

    2014-04-16

    In aphids, clonal individuals can show distinct morphologic traits in response to environmental cues. Such phenotypic plasticity cannot be studied with classical genetic model organisms such as Caenorhabditis elegans or Drosophila melanogaster. The genetic basis of this biological process remain unknown, as mutations affecting this process are not available in aphids. Here, we describe a protocol to treat third-stage larvae with an alkylating mutagen, ethyl methanesulfonate (EMS), to generate random mutations within the Acyrthosiphon pisum genome. We found that even low concentrations of EMS were toxic for two genotypes of A. pisum. Mutagenesis efficiency was nevertheless assessed by estimating the occurrence of mutational events on the X chromosome. Indeed, any lethal mutation on the X-chromosome would kill males that are haploid on the X so that we used the proportion of males as an estimation of mutagenesis efficacy. We could assess a putative mutation rate of 0.4 per X-chromosome at 10 mM of EMS. We then applied this protocol to perform a small-scale mutagenesis on parthenogenetic individuals, which were screened for defects in their ability to produce sexual individuals in response to photoperiod shortening. We found one mutant line showing a reproducible altered photoperiodic response with a reduced production of males and the appearance of aberrant winged males (wing atrophy, alteration of legs morphology). This mutation appeared to be stable because it could be transmitted over several generations of parthenogenetic individuals. To our knowledge, this study represents the first example of an EMS-generated aphid mutant.

  13. EMS Mutagenesis in the Pea Aphid Acyrthosiphon pisum

    PubMed Central

    Tagu, Denis; Le Trionnaire, Gaël; Tanguy, Sylvie; Gauthier, Jean-Pierre; Huynh, Jean-René

    2014-01-01

    In aphids, clonal individuals can show distinct morphologic traits in response to environmental cues. Such phenotypic plasticity cannot be studied with classical genetic model organisms such as Caenorhabditis elegans or Drosophila melanogaster. The genetic basis of this biological process remain unknown, as mutations affecting this process are not available in aphids. Here, we describe a protocol to treat third-stage larvae with an alkylating mutagen, ethyl methanesulfonate (EMS), to generate random mutations within the Acyrthosiphon pisum genome. We found that even low concentrations of EMS were toxic for two genotypes of A. pisum. Mutagenesis efficiency was nevertheless assessed by estimating the occurrence of mutational events on the X chromosome. Indeed, any lethal mutation on the X-chromosome would kill males that are haploid on the X so that we used the proportion of males as an estimation of mutagenesis efficacy. We could assess a putative mutation rate of 0.4 per X-chromosome at 10 mM of EMS. We then applied this protocol to perform a small-scale mutagenesis on parthenogenetic individuals, which were screened for defects in their ability to produce sexual individuals in response to photoperiod shortening. We found one mutant line showing a reproducible altered photoperiodic response with a reduced production of males and the appearance of aberrant winged males (wing atrophy, alteration of legs morphology). This mutation appeared to be stable because it could be transmitted over several generations of parthenogenetic individuals. To our knowledge, this study represents the first example of an EMS-generated aphid mutant. PMID:24531730

  14. Emulsifying and foaming properties of commercial yellow pea (Pisum sativum L.) seed flours.

    PubMed

    Aluko, Rotimi E; Mofolasayo, Olawunmi A; Watts, Beverley M

    2009-10-28

    Commercial yellow pea seed flours prepared by a patented wet-milling process and pea protein isolate (PPI) were analyzed for emulsifying and foaming properties at pH 3.0, 5.0, and 7.0 and compared to soybean protein isolate (SPI). PPI and SPI formed emulsions with significantly smaller (p < 0.05) oil droplet sizes, 16-30 and 23-54 microm, respectively, than flours that primarily contained fiber such as Centara III and IV, or those that consisted mainly of starch: Centu-tex, Uptake 80 and Accu-gel. PPI was a better emulsifier than SPI at pH 7.0, and a better foaming agent at pH 3.0 and pH 7.0, although foaming capacity varied with sample concentration. Centu-tex and Uptake 80 have exactly the same chemical composition, but the latter has a much smaller flour particle size range, and had significantly smaller (p < 0.05) emulsion oil droplets. Incorporation of pea starch into SPI emulsions produced a synergistic effect that led to significant increases (p < 0.05) in emulsification capacity (reduced emulsion oil droplet size) when compared to SPI or starch alone. These results showed that PPI had generally significantly higher (p < 0.05) emulsion and foam forming properties than SPI, and that pea starch could be used to improve the quality of SPI-stabilized food emulsions.

  15. Auxin regulation of a proton translocating ATPase in pea root plasma membrane vesicles. [Pisum sativum. L

    SciTech Connect

    Gabathuler, R.; Cleland, R.E.

    1985-12-01

    Pea root microsomal vesicles have been fractionated on a Dextran step gradient to give three fractions, each of which carries out ATP-dependent proton accumulation as measured by fluorescence quenching of quinacrine. The fraction at the 4/6% Dextran interface is enriched in plasma membrane, as determined by UDPG sterol glucosyltransferase and vanadate-inhibited ATPase. The vanadate-sensitive phosphohydrolase is not specific for ATP, has a K/sub m/ of about 0.23 millimolar for MgATP, is only slightly affected by K/sup +/ or Cl/sup -/ and is insensitive to auxin. Proton transport, on the other hand, is more specific for ATP, enhanced by anions (NO/sub 3//sup -/ > Cl/sup -/) and has a K/sub m/ of about 0.7 millimolar. Auxins decrease the K/sub m/ to about 0.35 millimolar, with no significant effect on the V/sub max/, while antiauxins or weak acids have no such effect. It appears that auxin has the ability to alter the efficiency of the ATP-driven proton transport.

  16. Remediation of cadmium toxicity in field peas (Pisum sativum L.) through exogenous silicon.

    PubMed

    Rahman, Mohammad Farhadur; Ghosal, Anubrata; Alam, Mohammad Firoz; Kabir, Ahmad Humayan

    2017-01-01

    Cadmium (Cd) is an important phytotoxic element causing health hazards. This work investigates whether and how silicon (Si) influences the alleviation of Cd toxicity in field peas at biochemical and molecular level. The addition of Si in Cd-stressed plants noticeably increased growth and development as well as total protein and membrane stability of Cd-stressed plants, suggesting that Si does have critical roles in Cd detoxification in peas. Furthermore, Si supplementation in Cd-stressed plants showed simultaneous significant increase and decrease of Cd and Fe in roots and shoots, respectively, compared with Cd-stressed plants. At molecular level, GSH1 (phytochelatin precursor) and MTA (metallothionein) transcripts predominantly expressed in roots and strongly induced due to Si supplementation in Cd-stressed plants compared with Cd-free conditions, suggesting that these chelating agents may bind to Cd leading to vacuolar sequestration in roots. Furthermore, pea Fe transporter (RIT1) showed downregulation in shoots when plants were treated with Si along with Cd compared with Cd-treated conditions. It is consistent with the physiological observations and supports the conclusion that alleviation of Cd toxicity in pea plants might be associated with Cd sequestration in roots and reduced Cd translocation in shoots through the regulation of Fe transport. Furthermore, increased CAT, POD, SOD and GR activity along with elevated S-metabolites (cysteine, methionine, glutathione) implies the active involvement of ROS scavenging and plays, at least in part, to the Si-mediated alleviation of Cd toxicity in pea. The study provides first mechanistic evidence on the beneficial effect of Si on Cd toxicity in pea plants. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Effectiveness of rhizobacteria containing ACC deaminase for growth promotion of peas (Pisum sativum) under drought conditions.

    PubMed

    Zahir, Z A; Munir, A; Asghar, H N; Shaharoona, B; Arshad, M

    2008-05-01

    A series of experiments were conducted to assess the effectiveness of rhizobacteria containing 1-aminocyclopropane- 1-carboxylate (ACC) deaminase for growth promotion of peas under drought conditions. Ten rhizobacteria isolated from the rhizosphere of different crops (peas, wheat, and maize) were screened for their growth promoting ability in peas under axenic condition. Three rhizobacterial isolates, Pseudomonas fluorescens biotype G (ACC-5), P. fluorescens (ACC-14), and P. putida biotype A (Q-7), were selected for pot trial on the basis of their source, ACC deaminase activity, root colonization, and growth promoting activity under axenic conditions. Inoculated and uninoculated (control) seeds of pea cultivar 2000 were sown in pots (4 seeds/pot) at different soil moisture levels (25, 50, 75, and 100% of field capacity). Results revealed that decreasing the soil moisture levels from 100 to 25% of field capacity significantly decreased the growth of peas. However, inoculation of peas with rhizobacteria containing ACC deaminase significantly decreased the "drought stress imposed effects" on growth of peas, although with variable efficacy at different moisture levels. At the lowest soil moisture level (25% field capacity), rhizobacterial isolate Pseudomonas fluorescens biotype G (ACC-5) was found to be more promising compared with the other isolates, as it caused maximum increases in fresh weight, dry weight, root length, shoot length, number of leaves per plant, and water use efficiency on fresh and dry weight basis (45, 150, 92, 45, 140, 46, and 147%, respectively) compared with respective uninoculated controls. It is highly likely that rhizobacteria containing ACC deaminase might have decreased the drought-stress induced ethylene in inoculated plants, which resulted in better growth of plants even at low moisture levels. Therefore, inoculation with rhizobacteria containing ACC deaminase could be helpful in eliminating the inhibitory effects of drought stress on the growth of peas.

  18. Breeding approaches for crenate broomrape (Orobanche crenata Forsk.) management in pea (Pisum sativum L.).

    PubMed

    Rubiales, Diego; Fernández-Aparicio, Monica; Pérez-de-Luque, Alejandro; Castillejo, Mari A; Prats, Elena; Sillero, Josefina C; Rispail, Nicolas; Fondevilla, Sara

    2009-05-01

    Pea cultivation is strongly hampered in Mediterranean and Middle East farming systems by the occurrence of Orobanche crenata Forsk. Strategies of control have been developed, but only marginal successes have been achieved. Most control methods are either unfeasible, uneconomical, hard to achieve or result in incomplete protection. The integration of several control measures is the most desirable strategy. [corrected] Recent developments in control are presented and re-evaluated in light of recent developments in crop breeding and molecular genetics. These developments are placed within a framework that is compatible with current agronomic practices. The current focus in applied breeding is leveraging biotechnological tools to develop more and better markers to speed up the delivery of improved cultivars to the farmer. To date, however, progress in marker development and delivery of useful markers has been slow. The application of knowledge gained from basic genomic research and genetic engineering will contribute to more rapid pea improvement for resistance against O. crenata and/or the herbicide.

  19. Pea starch (Pisum sativum L.) with slow digestion property produced using β-amylase and transglucosidase.

    PubMed

    Shi, Miaomiao; Zhang, Zhiheng; Yu, Shujuan; Wang, Kai; Gilbert, Robert G; Gao, Qunyu

    2014-12-01

    Starches extracted from wrinkled (WP) and smooth (SP) peas were treated using β-amylase (B) alone and also with a combination of β-amylase and transglucosidase (BT). After enzymatic treatment, the proportions of slowly digested starch in WP-B, WP-BT, SP-B and SP-BT samples were increased by 6%, 9%, 9% and 12%, respectively. Starches treated by a combination of β-amylase and transglucosidase exhibited a smaller amount of longer amylopectin chains, a larger amount of short amylopectin chains, and higher branching fraction. The branching fraction was significantly increased, with an increase of 8%, 10%, 13% and 14% for WP-B, WP-BT, SP-B and SP-BT, respectively. The maximum absorbance and iodine binding of enzyme-treated starches were reduced compared with their native starch parents. The C-type crystalline structure completely disappeared after enzymatic treatment. The results support previous findings that increases in the amount of shorter amylopectin chains and branch fraction are likely to contribute to the slow digestion of starch. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. The effect of UV-B radiation on chloroplast translation in Pisum sativum

    SciTech Connect

    Raab, M.M.; Jagendorf, A.T. )

    1990-05-01

    UV-B radiation has previously been reported to reduce growth, flowering, and net photosynthesis. The present study examines the effect of UV-B radiation on isolated chloroplast of 7-10 day old pea seedlings. Amount of ({sup 3}H)-Leu incorporated into isolated chloroplasts was measured in the presence or absence of UV-B exposure. Preliminary experiments show a 30% inhibition of protein synthesis in isolated chloroplasts after only 20 mins of UV-B exposure (6.9 J/m{sup 2}/30 min). Percent inhibition of chloroplast translation is directly correlated with UV-B exposure over a 60 min time span. Preliminary studies also show no change in both cold and radiolabeled protein profiles as expressed on 1-D PAGE and autofluorography. Comparative studies on the sensitivity of e{sup {minus}} flow vs protein synthesis following UV-B exposure are underway. Further work on the role of oxygen free radicals and the specific site of action of UV-B damage to the translation machinery of chloroplasts will be discussed.

  1. Isolation and characterization of NADP+-linked isocitrate dehydrogenase of germinating pea seeds (Pisum sativum).

    PubMed

    Srivastava, P K; Singh, D S

    2001-10-01

    NADP+-linked isocitrate dehydrogenase (E.C.1.1.1.42) has been purified to homogeneity from germinating pea seeds. The enzyme is a tetrameric protein (mol wt, about 146,000) made up of apparently identical monomers (subunit mol wt, about 36,000). Thermal inactivation of purified enzyme at 45 degrees and 50 degrees C shows simple first order kinetics. The enzyme shows optimum activity at pH range 7.5-8. Effect of substrate [S] on enzyme activity at different pH (6.5-8) suggests that the proton behaves formally as an "uncompetitive inhibitor". A basic group of the enzyme (site) is protonated in this pH range in the presence of substrate only, with a pKa equal to 6.78. On successive dialysis against EDTA and phosphate buffer, pH 7.8 at 0 degrees C, yields an enzymatically inactive protein showing kinetics of thermal inactivation identical to the untreated (native) enzyme. Maximum enzyme activity is observed in presence of Mn2+ and Mg2+ ions (3.75 mM). Addition of Zn2+, Cd2+, Co2+ and Ca2+ ions brings about partial recovery. Other metal ions Fe2+, Cu2+ and Ni2+ are ineffective.

  2. Functional Properties of Pea (Pisum sativum, L.) Protein Isolates Modified with Chymosin

    PubMed Central

    Barać, Miroljub; Čabrilo, Slavica; Pešić, Mirjana; Stanojević, Slađana; Pavlićević, Milica; Maćej, Ognjen; Ristić, Nikola

    2011-01-01

    In this paper, the effects of limited hydrolysis on functional properties, as well as on protein composition of laboratory-prepared pea protein isolates, were investigated. Pea protein isolates were hydrolyzed for either 15, 30 and 60 min with recombined chymosin (Maxiren). The effect of enzymatic action on solubility, emulsifying and foaming properties at different pH values (3.0; 5.0; 7.0 and 8.0) was monitored. Chymosin can be a very useful agent for improvement of functional properties of isolates. Action of this enzyme caused a low degree of hydrolysis (3.9–4.7%), but improved significantly functional properties of pea protein isolates (PPI), especially at lower pH values (3.0–5.0). At these pH values all hydrolysates had better solubility, emulsifying activity and foaming stability, while longer-treated samples (60 min) formed more stable emulsions at higher pH values (7.0, 8.0) than initial isolates. Also, regardless of pH value, all hydrolysates showed improved foaming ability. A moderate positive correlation between solubility and emulsifying activity index (EAI) (0.74) and negative correlation between solubility and foam stability (−0.60) as well as between foam stability (FS) and EAI (−0.77) were observed. Detected enhancement in functional properties was a result of partial hydrolysis of insoluble protein complexes. PMID:22272078

  3. Symbiotic activity of pea (Pisum sativum) after application of Nod factors under field conditions.

    PubMed

    Siczek, Anna; Lipiec, Jerzy; Wielbo, Jerzy; Kidaj, Dominika; Szarlip, Paweł

    2014-04-29

    Growth and symbiotic activity of legumes are mediated by Nod factors (LCO, lipo-chitooligosaccharides). To assess the effects of application of Nod factors on symbiotic activity and yield of pea, a two-year field experiment was conducted on a Haplic Luvisol developed from loess. Nod factors were isolated from Rhizobium leguminosarum bv. viciae strain GR09. Pea seeds were treated with the Nod factors (10⁻¹¹ M) or water (control) before planting. Symbiotic activity was evaluated by measurements of nitrogenase activity (acetylene reduction assay), nodule number and mass, and top growth by shoot mass, leaf area, and seed and protein yield. Nod factors generally improved pea yield and nitrogenase activity in the relatively dry growing season 2012, but not in the wet growing season in 2013 due to different weather conditions.

  4. S-Nitrosylated proteins in pea (Pisum sativum L.) leaf peroxisomes: changes under abiotic stress

    PubMed Central

    Ortega-Galisteo, Ana P.; Rodríguez-Serrano, María; Pazmiño, Diana M.; Gupta, Dharmendra K.; Sandalio, Luisa M.; Romero-Puertas, María C.

    2012-01-01

    Peroxisomes, single-membrane-bounded organelles with essentially oxidative metabolism, are key in plant responses to abiotic and biotic stresses. Recently, the presence of nitric oxide (NO) described in peroxisomes opened the possibility of new cellular functions, as NO regulates diverse biological processes by directly modifying proteins. However, this mechanism has not yet been analysed in peroxisomes. This study assessed the presence of S-nitrosylation in pea-leaf peroxisomes, purified S-nitrosylated peroxisome proteins by immunoprecipitation, and identified the purified proteins by two different mass-spectrometry techniques (matrix-assisted laser desorption/ionization tandem time-of-flight and two-dimensional nano-liquid chromatography coupled to ion-trap tandem mass spectrometry). Six peroxisomal proteins were identified as putative targets of S-nitrosylation involved in photorespiration, β-oxidation, and reactive oxygen species detoxification. The activity of three of these proteins (catalase, glycolate oxidase, and malate dehydrogenase) is inhibited by NO donors. NO metabolism/S-nitrosylation and peroxisomes were analysed under two different types of abiotic stress, i.e. cadmium and 2,4-dichlorophenoxy acetic acid (2,4-D). Both types of stress reduced NO production in pea plants, and an increase in S-nitrosylation was observed in pea extracts under 2,4-D treatment while no total changes were observed in peroxisomes. However, the S-nitrosylation levels of catalase and glycolate oxidase changed under cadmium and 2,4-D treatments, suggesting that this post-translational modification could be involved in the regulation of H2O2 level under abiotic stress. PMID:22213812

  5. Quality traits analysis and protein profiling of field pea (Pisum sativum) germplasm from Himalayan region.

    PubMed

    Sharma, Shagun; Singh, Narpinder; Virdi, Amardeep Singh; Rana, Jai Chand

    2015-04-01

    The grain and flour characteristics of different field pea (FP) accessions were evaluated. Accessions with higher grain weight had less compact structure with a greater proportion of large-sized starch granules. Accessions with higher protein content had lower starch content, blue value and λ(max) whereas accessions with higher amylose showed higher resistant starch (RS) and final viscosity and lower rapidly digestible starch (RDS). Ca, Zn, K and Fe content vary significantly amongst different accessions and creamish green and white seeds accessions showed higher Fe and Zn content. Yellow coloured accessions (1.36-3.71%) showed lower antioxidant activity as compared to brownish and green coloured accessions (4.06-9.30%). Out of 21 major polypeptides observed (9-100 kDa), 11 showed differential trypsin inhibitory activity (TIA) under non-reducing conditions. Polypeptides of 68, 46, 33 and 22 kDa showed prominent TIA.

  6. Extrachromosomal DNA and cell differentiation in cultured pea roots (Pisum sativum)

    SciTech Connect

    Van't Hof, J.

    1986-01-01

    Histological, cytological and molecular analyses of DNA replication and differentiation and meristematic precursors of vascular parenchyma in cultured pea roots show that the initial steps in the transition from a dividing to a differentiated cell involve retardation of DNA replication in late S phase and production of extrachromosomal molecules by excision from late replicating chromosomal DNA. Portions of the extrachromosomal DNA are displaced by a strand displacement mechanism thereby producing free single-stranded molecules. That extrachromosomal molecules have rDNA reflects the fact that in pea roots replication of the ribosomal genes is maximal during late S phase (unpublished results). It is likely, therefore, that the excised late replicating DNA contains some rDNA sequences. 30 refs., 3 figs.

  7. Relating physico-chemical properties of frozen green peas (Pisum sativum L.) to sensory quality.

    PubMed

    Nleya, Kathleen M; Minnaar, Amanda; de Kock, Henriëtte L

    2014-03-30

    The acceptability of frozen green peas depends on their sensory quality. There is a need to relate physico-chemical parameters to sensory quality. In this research, six brands of frozen green peas representing product sold for retail and caterer's markets were purchased and subjected to descriptive sensory evaluation and physico-chemical analyses (including dry matter content, alcohol insoluble solids content, starch content, °Brix, residual peroxidase activity, size sorting, hardness using texture analysis and colour measurements) to assess and explain product quality. The sensory quality of frozen green peas, particularly texture properties, were well explained using physico-chemical methods of analysis notably alcohol insoluble solids, starch content, hardness and °Brix. Generally, retail class peas were of superior sensory quality to caterer's class peas although one caterer's brand was comparable to the retail brands. Retail class peas were sweeter, smaller, greener, more moist and more tender than the caterer's peas. Retail class peas also had higher °Brix, a(*) , hue and chroma values; lower starch, alcohol insoluble solids, dry matter content and hardness measured. The sensory quality of frozen green peas can be partially predicted by measuring physico-chemical parameters particularly °Brix and to a lesser extent hardness by texture analyser, alcohol insoluble solids, dry matter and starch content. © 2013 Society of Chemical Industry.

  8. Nutritional evaluation of low-phytate peas (Pisum sativum L.) for young broiler chicks.

    PubMed

    Thacker, Philip; Deep, Aman; Petri, Daniel; Warkentin, Thomas

    2013-02-01

    This experiment determined the effects of including normal and low-phytate peas in diets fed to young broiler chickens on performance, phosphorus availability and bone strength. A total of 180, day-old, male broilers (Ross-308 line) were assigned to six treatments. The control was based on corn and soybean meal while two additional corn-based diets were formulated containing 30% of either normal or low-phytate pea providing 0.45% available phosphorus. For each of these three diets, a similar diet was formulated by reducing the amount of dicalcium phosphate to produce a diet with 0.3% available phosphorus. The total tract apparent availability (TTAA) of phosphorus was higher (p = 0.02) for broilers fed the low-phytate pea than for birds fed the normal pea diets. Birds fed diets containing the lower level of phosphorus had a higher TTAA of phosphorus (50.64 vs. 46.68%) than broilers fed diets adequate in phosphorus. Protein source had no effect on weight gain, feed intake or feed conversion. Broilers fed the low phosphorus diets had lower weight gain (p = 0.04) and feed intake (p < 0.01) than broilers fed the higher phosphorus level. Bone strength was higher (p < 0.01) for broilers fed diets based on low-phytate pea than for those fed diets based on normal pea or soybean meal. Increasing the availability of the phosphorus in peas could mean that less inorganic phosphorus would be required in order to meet the nutritional requirements of broilers. Since inorganic phosphorus sources tend to be expensive, a reduction in their use would lower ration costs. In addition, increased availability of phosphorus would reduce the amount of phosphorus excreted thus reducing the amount of phosphorus that can potentially pollute the environment.

  9. Alterations in antioxidant metabolism and associated enzymes in pea (Pisum sativum) exposed to sulfur dioxide

    SciTech Connect

    Madamanchi, N.R.; Alscher, R.G. )

    1990-05-01

    The response of glutathione and ascorbate and the enzymes glutathione reductase (GR) and superoxide dismutase (SOD) was studied in two cultivars of pea known to be differentially sensitive to SO{sub 2} (0.8 ppm). Total glutathione accumulated more rapidly on exposure to SO{sub 2} in insensitive cultivar Progress compared to the sensitive cultivar Nugget, confirming our previous results. However, corresponding changes in oxidized glutathione were not observed and ascorbate levels did not change over the course of the exposure. Changes in the activity of GR corresponded to the changes in total glutatione levels. Preliminary results indicate that SOD activity increased to a significantly higher extent in Progress than in Nugget. These data suggest a significant role for GR and possibly SOD in resistance to oxidative stress.

  10. Comparative Transcriptomic Analyses of Vegetable and Grain Pea (Pisum sativum L.) Seed Development

    PubMed Central

    Liu, Na; Zhang, Guwen; Xu, Shengchun; Mao, Weihua; Hu, Qizan; Gong, Yaming

    2015-01-01

    Understanding the molecular mechanisms regulating pea seed developmental process is extremely important for pea breeding. In this study, we used high-throughput RNA-Seq and bioinformatics analyses to examine the changes in gene expression during seed development in vegetable pea and grain pea, and compare the gene expression profiles of these two pea types. RNA-Seq generated 18.7 G of raw data, which were then de novo assembled into 77,273 unigenes with a mean length of 930 bp. Our results illustrate that transcriptional control during pea seed development is a highly coordinated process. There were 459 and 801 genes differentially expressed at early and late seed maturation stages between vegetable pea and grain pea, respectively. Soluble sugar and starch metabolism related genes were significantly activated during the development of pea seeds coinciding with the onset of accumulation of sugar and starch in the seeds. A comparative analysis of genes involved in sugar and starch biosynthesis in vegetable pea (high seed soluble sugar and low starch) and grain pea (high seed starch and low soluble sugar) revealed that differential expression of related genes at late development stages results in a negative correlation between soluble sugar and starch biosynthetic flux in vegetable and grain pea seeds. RNA-Seq data was validated by using real-time quantitative RT-PCR analysis for 30 randomly selected genes. To our knowledge, this work represents the first report of seed development transcriptomics in pea. The obtained results provide a foundation to support future efforts to unravel the underlying mechanisms that control the developmental biology of pea seeds, and serve as a valuable resource for improving pea breeding. PMID:26635856

  11. Changes of the oligomeric structure of legumin from pea (Pisum sativum L.) after succinylation.

    PubMed

    Schwenke, K D; Zirwer, D; Gast, K; Görnitz, E; Linow, K J; Gueguen, J

    1990-12-12

    The influence of various levels of succinylation on the structure of the legumin from pea seed has been studied by the techniques of sedimentation velocity, viscometry, fluorescence and circular dichroism spectroscopy, as well as dynamic light scattering. The protein dissociates gradually into the 3S subunit forming a 7S intermediate. At a level of 75-80% succinylation, sudden unfolding of the protein occurs characterized by drastic changes in viscometric and spectroscopic properties. The fluorescence spectra point to the formation of a novel organized structure at a moderate degree of modification before the molecular unfolding takes place. The succinylated subunit was shown to have a sedimentation coefficient of 3.2S, a diffusion coefficient of 5.03 x 10(-7) cm2 . s-1 a Stokes' radius of 4.24 nm, a partial specific volume of 0.703 ml/g, an intrinsic viscosity of 0.13 dl/g, a molar mass of 52.2 kDa and a frictional ratio of 1.74.

  12. Physicochemical properties and in vitro digestibility of flour and starch from pea (Pisum sativum L.) cultivars.

    PubMed

    Chung, Hyun-Jung; Liu, Qiang

    2012-01-01

    Flours and isolated starches from three different cultivars (1544-8, 1658-11 and 1760-8) of pea grown under identical environmental conditions were evaluated for their physicochemical properties and in vitro digestibility. The protein content, total starch content and apparent amylose content of pea flour ranged from 24.4 to 26.3%, 48.8 to 50.2%, and 13.9 to 16.7%, respectively. In pea starches, the 1760-8 showed higher apparent amylose content and total starch content than the other cultivars. Pea starch granules were irregularly shaped, ranging from oval to round with a smooth surface. All pea starches showed C-type X-ray diffraction pattern with relative crystallinity ranging between 23.7 and 24.7%. Pea starch had only a single endothermic transition (12.1-14.2 J/g) in the DSC thermogram, whereas pea flour showed two separate endothermic transitions corresponding to starch gelatinization (4.54-4.71 J/g) and disruption of the amylose-lipid complex (0.36-0.78 J/g). In pea cultivars, the 1760-8 had significantly higher setback and final viscosity than the other cultivars in both pea flour (672 and 1170cP, respectively) and isolated starch (2901 and 4811cP). The average branch chain length of pea starches ranged from 20.1 to 20.3. The 1760-8 displayed a larger proportion of short branch chains, DP (degree of polymerization) 6-12 (21.1%), and a smaller proportion of long branch chains, DP≥37 (8.4%). The RDS, SDS and RS contents of pea flour ranged from 23.7 to 24.1%, 11.3 to 12.8%, and 13.2 to 14.8%, respectively. In pea starches, the 1760-8 showed a lower RDS content but higher SDS and RS contents. The expected glycemic index (eGI), based on the hydrolysis index, ranged from 36.9 to 37.7 and 69.8 to 70.7 for pea flour and isolated pea starch, respectively.

  13. Symbiotic Activity of Pea (Pisum sativum) after Application of Nod Factors under Field Conditions

    PubMed Central

    Siczek, Anna; Lipiec, Jerzy; Wielbo, Jerzy; Kidaj, Dominika; Szarlip, Paweł

    2014-01-01

    Growth and symbiotic activity of legumes are mediated by Nod factors (LCO, lipo-chitooligosaccharides). To assess the effects of application of Nod factors on symbiotic activity and yield of pea, a two-year field experiment was conducted on a Haplic Luvisol developed from loess. Nod factors were isolated from Rhizobium leguminosarum bv. viciae strain GR09. Pea seeds were treated with the Nod factors (10−11 M) or water (control) before planting. Symbiotic activity was evaluated by measurements of nitrogenase activity (acetylene reduction assay), nodule number and mass, and top growth by shoot mass, leaf area, and seed and protein yield. Nod factors generally improved pea yield and nitrogenase activity in the relatively dry growing season 2012, but not in the wet growing season in 2013 due to different weather conditions. PMID:24786094

  14. Mapping QTL for Fusarium wilt Race 2 partial resistance in pea (Pisum sativum L.)

    USDA-ARS?s Scientific Manuscript database

    Fusarium oxysporum f.sp. pisi (Fop) is present in pea production regions worldwide, and causes a vascular wilt resulting in significant crop losses. Four races of Fop have been identified and resistance to each reportedly conferred by an individual single dominant gene. Fnw confers resistance to Fop...

  15. Label-free quantitative proteomic analysis of tolerance to drought in Pisum sativum.

    PubMed

    Castillejo, María-Ángeles; Iglesias-García, Rebeca; Wienkoop, Stefanie; Rubiales, Diego

    2016-11-01

    Abiotic stresses caused by adverse environmental conditions are responsible for heavy economic losses on pea crop, being drought one of the most important abiotic constraints. Development of pea cultivars well adapted to dry conditions has been one of the major tasks in breeding programs. The increasing food requirements drive the necessity to broaden the molecular basis of tolerance to drought to develop pea cultivars well adapted to dry conditions. We have used a shotgun proteomic approach (nLC-MSMS) to study the tolerance to drought in three pea genotypes that were selected based on differences in the level of water deficit tolerance. Multivariate statistical analysis of data unraveled 367 significant differences of 700 identified when genotypes and/or treatment were compared. More than half of the significantly changed proteins belong to primary metabolism and protein regulation categories. We propose different mechanisms to cope drought in the genotypes studied. Maintenance of the primary metabolism and protein protection seems a strategy for drought tolerance. On the other hand susceptibility might be related to maintenance of the homeostatic equilibrium, a very energy consuming process. Data are available via ProteomeXchange with identifier PXD004587. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Cells of pea (Pisum sativum) that differentiate from G2 phase have extrachromosomal DNA

    SciTech Connect

    Van't Hof, J.; Bjerknes, C.A.

    1982-04-01

    Velocity sedimentation in an alkaline sucrose gradient of newly replicated chromosomal DNA revealed the presence of extrachromosomal DNA that was not replicated by differentiating cells in the elongation zone. The extrachromosomal DNA had a number average molecular weight of 12 x 10/sup 6/ to 15 x 10/sup 6/ and a weight average molecular weight of 25 x 10/sup 6/, corresponding to about 26 x 10/sup 6/ and 50 x 10/sup 6/ daltons, respectively, of double-stranded DNA. The molecules were stable, lasting at least 72 h after being formed. Concurrent measurements by velocity sedimentation, autoradiography, and cytophotometry of isolated nuclei indicated that the extrachromosomal molecules were associated with root-tip-cells that stopped dividing and differentiated from G2 phase but not with those that stopped dividing and differentiated from G1 phase.

  17. Brassinosteroid-induced changes of lipid composition in leaves of Pisum sativum L. during senescence.

    PubMed

    Fedina, Evgenia; Yarin, Andrey; Mukhitova, Faimya; Blufard, Alexander; Chechetkin, Ivan

    2017-01-01

    The effect of steroid phytohormone 24-epibrassinolide (EBR) on the composition of some lipid classes (free fatty acids, triacylglycerols and galactolipids) in detached pea leaves was studied for the first time. EBR (0.1μM) promoted senescence and increased the content of 14:0, 16:0 and 18:1 free fatty acids as well as 18:2 and 18:3 bound to triacylglycerols in the detached leaves in contrast to mock-treated leaves. The content of all identified fatty acids bound to galactolipids decreased in the detached leaves treated with EBR compared to that in mock-treated leaves. These findings suggest that free fatty acids are liberated from polar lipids and then undergo esterification to neutral lipids in the detached leaves upon EBR treatment. We propose that steroid phytohormones may be involved into regulation of leaf senescence via alteration of cell lipid composition. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Synteny analysis of loci controlling partial resistance to Aphanomyces euteiches between Pisum sativum and Medicago truncatula

    USDA-ARS?s Scientific Manuscript database

    Aphanomyces root rot, due to Aphanomyces euteiches, is one of the most damaging diseases of pea worldwide. Breeding for partial polygenic resistance to Aphanomyces root rot is a major objective for the development of the pea crop in Europe. Our objectives were to study i)- the diversity of resistanc...

  19. Extrachromosomal DNA of pea (Pisum sativum) root-tip cells replicates by strand displacement

    SciTech Connect

    Krimer, D.B.; Van't Hof, J.

    1983-04-01

    In cultured pea roots there is extrachromosomal DNA associated with cells that differentiate from the G/sub 2/ phase of the cell cycle that is absent from those that differentiate from the G/sub 1/ phase. The authors examined this extrachromosomal DNA by electron microscopy and found that it consisted of three types: (i) double-stranded linear molecules with single-stranded branches (74%), (ii) double-stranded molecules without branches (26%), and (iii) free single-stranded molecules. The double-stranded molecules with or without branches were similar in length, having a modal length of 10-15 ..mu..m. The free single-stranded molecules were shorter and had a mean length of 3.8 ..mu..m. The length of the branches attached to the duplex molecules was only slightly less than that of the free form. The duplex molecules with branches were interpreted as configurations reflecting an ongoing strand-displacement process that results in free single-stranded molecules. Finally, measurements on duplex molecules with multiple branches suggested that the extrachromosomal DNA may exist in the form of tandemly repeated sequences. 8 references, 8 figures.

  20. Mitogen-activated protein kinases participate in determination of apical-basal symmetry in Pisum sativum.

    PubMed

    Winnicki, Konrad; Polit, Justyna Teresa; Żabka, Aneta; Maszewski, Janusz

    2017-03-01

    Mitogen-activated protein kinases (MAPKs) are implicated in various processes in plants. Apart from response to biotic and abiotic stresses they are involved in regulation of embryo development. Although MAPKs were found to be indispensable during embryo development it has never been established at which stages of embryogenesis and in which regions of a plant embryo activated MAPKs can be observed. Here, we show that apical and basal regions display activation of the same types of MAPKs and the only difference concerns the level of their phosphorylation and cellular localization. Dually-phosphorylated MAPKs were found in nuclei of the apical region of an embryo both at the early and late cotyledonary stage while no immunofluorescence signals were detected in nuclei of the basal region. However, in this case phosphorylated MAPKs were immunodetected in cytoplasm in the apical domain of cortex cells, indicating their role in auxin transport from the basal to apical region of an embryo. Furthermore, obtained data indicate that nuclear localization of activated MAPKs may result from epigenetic modifications and polar auxin transport. The presented data and previous studies lead to the conclusion that activated MAPKs and their cellular localization define apical and basal regions during formation of an apical-basal axis. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. A rapid method to increase the number of F₁ plants in pea (Pisum sativum) breeding programs.

    PubMed

    Espósito, M A; Almirón, P; Gatti, I; Cravero, V P; Anido, F S L; Cointry, E L

    2012-08-16

    In breeding programs, a large number of F₂ individuals are required to perform the selection process properly, but often few such plants are available. In order to obtain more F₂ seeds, it is necessary to multiply the F₁ plants. We developed a rapid, efficient and reproducible protocol for in vitro shoot regeneration and rooting of seeds using 6-benzylaminopurine. To optimize shoot regeneration, basic medium contained Murashige and Skoog (MS) salts with or without B5 Gamborg vitamins and different concentrations of 6-benzylaminopurine (25, 50 and 75 μM) using five genotypes. We found that modified MS (B5 vitamins + 25 μM 6-benzylaminopurine) is suitable for in vitro shoot regeneration of pea. Thirty-eight hybrid combinations were transferred onto selected medium to produce shoots that were used for root induction on MS medium supplemented with α-naphthalene-acetic acid. Elongated shoots were developed from all hybrid genotypes. This procedure can be used in pea breeding programs and will allow working with a large number of plants even when the F₁ plants produce few seeds.

  2. Cloning and characterization of CBL-CIPK signalling components from a legume (Pisum sativum).

    PubMed

    Mahajan, Shilpi; Sopory, Sudhir K; Tuteja, Narendra

    2006-03-01

    The studies on calcium sensor calcineurin B-like protein (CBL) and CBL interacting protein kinases (CIPK) are limited to Arabidopsis and rice and their functional role is only beginning to emerge. Here, we present cloning and characterization of a protein kinase (PsCIPK) from a legume, pea, with novel properties. The PsCIPK gene is intronless and encodes a protein that showed partial homology to the members of CIPK family. The recombinant PsCIPK protein was autophosphorylated at Thr residue(s). Immunoprecipitation and yeast two-hybrid analysis showed direct interaction of PsCIPK with PsCBL, whose cDNA and genomic DNA were also cloned in this study. PsCBL showed homology to AtCBL3 and contained calcium-binding activity. We demonstrate for the first time that PsCBL is phosphorylated at its Thr residue(s) by PsCIPK. Immunofluorescence/confocal microscopy showed that PsCBL is exclusively localized in the cytosol, whereas PsCIPK is localized in the cytosol and the outer membrane. The exposure of plants to NaCl, cold and wounding co-ordinately upregulated the expression of PsCBL and PsCIPK genes. The transcript levels of both genes were also coordinately stimulated in response to calcium and salicylic acid. However, drought and abscisic acid had no effect on the expression of these genes. These studies show the ubiquitous presence of CBL/CIPK in higher plants and enhance our understanding of their role in abiotic and biotic stress signalling.

  3. Localization of Adenosine Triphosphatase Activity on the Chloroplast Envelope in Tendrils of Pisum sativum1

    PubMed Central

    Sabnis, Dinkar D.; Gordon, Mildred; Galston, Arthur W.

    1970-01-01

    When samples of pea tendril tissue were incubated in the Wachstein-Meisel medium for the demonstration of adenosine triphosphatases, deposits of lead reaction product were localized between the membranes of the chloroplast envelope. The presence of Mg2+ was necessary for adenosine triphosphatase activity, and Ca2+ could not substitute for this requirement. Varying the pH of incubation to 5.5 or 9.4 inhibited enzyme activity, as did the addition of p-chloromercuribenzoic acid or N-ethylmaleimide. The adenosine triphosphatase was apparently inactivated or degraded when the plants were grown in the dark for 24 hours prior to incubation. The enzyme was substrate-specific for adenosine triphosphate; no reaction was obtained with adenosine diphosphate, uridine triphosphate, inosine triphosphate, p-nitrophenyl phosphate, and sodium β-glycerophosphate. Sites of nonspecific depositions of lead are described. The adenosine triphosphatase on the chloroplast envelope may be involved in the light-induced contraction of this organelle. Images PMID:4245003

  4. Additional pea EST-SSR markers for comparative mapping in pea (Pisum sativum L.)

    USDA-ARS?s Scientific Manuscript database

    The availability of a large set of molecular markers is an excellent resource for both applied and basic research for any organism. The current consensus molecular map of pea is 1,430 cM and contains 239 simple sequence repeat markers. The goal was to identify and test unique gene-based simple sequ...

  5. Pea Mutants with Reduced Sensitivity to Far-Red Light Define an Important Role for Phytochrome A in Day-Length Detection.

    PubMed Central

    Weller, J. L.; Murfet, I. C.; Reid, J. B.

    1997-01-01

    In garden pea (Pisum sativum L.), a long-day plant, long photoperiods promote flowering by reducing the synthesis or transport of a graft-transmissible inhibitor of flowering. Previous physiological studies have indicated that this promotive effect is predominantly achieved through a response that requires long exposures to light and for which far-red (FR) light is the most effective. These characteristics implicate the action of phytochrome A (phyA). To investigate this matter further, we screened ethylmethane sulfonate-mutagenized pea seedlings for FR-unresponsive, potentially phyA-deficient mutants. Two allelic, recessive mutants were isolated and were designated fun1 for FR unresponsive. The fun1-1 mutant is specifically deficient in the PHYA apoprotein and has a seedling phenotype indistinguishable from wild type when grown under white light. However, fun1-1 plants grown to maturity under long photoperiods show a highly pleiotropic phenotype, with short internodes, thickened stems, delayed flowering and senescence, longer peduncles, and higher seed yield. This phenotype results in large part from an inability of fun1-1 to detect day extensions. These results establish a crucial role for phyA in the control of flowering in pea, and show that phyA mediates responses to both red and FR light. Furthermore, grafting and epistasis studies with fun1 and dne, a mutant deficient in the floral inhibitor, show that the roles of phyA in seedling deetiolation and in day-length detection are genetically separable and that the phyA-mediated promotion of flowering results from a reduction in the synthesis or transport of the floral inhibitor. PMID:12223768

  6. Phylogeny, phylogeography and genetic diversity of Pisum genus

    USDA-ARS?s Scientific Manuscript database

    Tribe Fabeae (formerly Vicieae) contains some of humanity's most important grain legume crops, namely Lathyrus; Lens; Pisum; Vicia and the monotypic genus Vavilovia. Our study based on molecular data, have positioned Pisum between Vicia and Lathyrus and being closely allied to Vavilovia. Study of p...

  7. Cannibalism in the pea aphid, Acyrthosiphon pisum.

    PubMed

    Cooper, Lucy C; Desjonqueres, Camille; Leather, Simon R

    2014-12-01

    Previous observations of cannibalism have been made in the aphid Acyrthosiphon pisum (L.): this article seeks to quantify factors contributing to such behaviors. We observed and quantified the responses of a number of clones and life stages to varying levels of starvation, in the form of increasingly desiccated Vica faba L. plants (receiving 50, 25, or 10 mL every second day) or a complete absence of host plant. We found that, while the longest incidences of cannibalism are carried out by juveniles (F = 3.45, P = 0.019, df = 3) and targeted at adults, the starvation treatments had the most significant effect on the prevalence of cannibalism in mature A. pisum (F = 2.24, P = 0.025, df = 9). Furthermore, there was no difference between the prevalence or duration of cannibalistic activities within and between different clones (P ≥ 0.05 in all cases), though juveniles were more likely to target unrelated aphids (V = 6 112, P = 0.011), and spent more time feeding on aphids from the same culture (V = 6 062, P = 0.018).

  8. Characterization by enzyme-linked immunosorbent assay of monoclonal antibodies to Pisum and Avena phytochrome

    SciTech Connect

    Cordonnier, M.M.; Greppin, H.; Pratt, L.H.

    1984-01-01

    Nine monoclonal antibodies to pea (Pisum sativum L.) and 16 to oat (Avena sativa L.) phytochrome are characterized by enzyme-linked immunosorbent assay against phytochrome from six different sources: pea, zucchini (Cucurbita pepo L.), lettuce (Lactuca sativa L.), oat, rye (Secale cereale L.), and barley (Hordeum vulgare L.). All antibodies were raised against phytochrome with a monomer size near 120,000 daltons. Nevertheless, none of them discriminated qualitatively between 118/114-kilodalton oat phytochrome and a photoreversible, 60-kilodalton proteolytic degradation product derived from it. In addition, none of the 23 antibodies tested discriminated substantially between phytochrome - red-absorbing form and phytochrome - far red-absorbing form. Two antibodies to pea and six to oat phytochrome also bound strongly to phytochrome from the other species, even though these two plants are evolutionarily widely divergent. Of these eight antibodies, two bound significantly to all of the six phytochrome preparations tested, indicating that these two may recognize highly conserved regions of the chromoprotein. Since the molecular function of phytochrome is unknown, these two antibodies may serve as unique probes for regions of this pigment that are important to its mode of action. 27 references, 3 figures, 1 table.

  9. Transcriptome of Dickeya dadantii Infecting Acyrthosiphon pisum Reveals a Strong Defense against Antimicrobial Peptides

    PubMed Central

    Costechareyre, Denis; Chich, Jean-François; Strub, Jean-Marc; Rahbé, Yvan; Condemine, Guy

    2013-01-01

    The plant pathogenic bacterium Dickeya dadantii has recently been shown to be able to kill the aphid Acyrthosiphon pisum. While the factors required to cause plant disease are now well characterized, those required for insect pathogeny remain mostly unknown. To identify these factors, we analyzed the transcriptome of the bacteria isolated from infected aphids. More than 150 genes were upregulated and 300 downregulated more than 5-fold at 3 days post infection. No homologue to known toxin genes could be identified in the upregulated genes. The upregulated genes reflect the response of the bacteria to the conditions encountered inside aphids. While only a few genes involved in the response to oxidative stress were induced, a strong defense against antimicrobial peptides (AMP) was induced. Expression of a great number of efflux proteins and transporters was increased. Besides the genes involved in LPS modification by addition of 4-aminoarabinose (the arnBCADTEF operon) and phosphoethanolamine (pmrC, eptB) usually induced in Gram negative bacteria in response to AMPs, dltBAC and pbpG genes, which confer Gram positive bacteria resistance to AMPs by adding alanine to teichoic acids, were also induced. Both types of modification confer D. dadantii resistance to the AMP polymyxin. A. pisum harbors symbiotic bacteria and it is thought that it has a very limited immune system to maintain these populations and do not synthesize AMPs. The arnB mutant was less pathogenic to A. pisum, which suggests that, in contrast to what has been supposed, aphids do synthesize AMP. PMID:23342088

  10. Analysis of apical hook formation in Alaska pea with a 3-D clinostat and agravitropic mutant ageotropum

    PubMed Central

    Miyamoto, Kensuke; Yamasaki, Takahiro; Uheda, Eiji; Ueda, Junichi

    2013-01-01

    The formation of the apical hook in dicotyledonous seedlings is believed to be effected by gravity in the dark. However, this notion is mostly based on experiments with the hook formed on the hypocotyl, and no detailed studies are available with the developmental manners of the hook, particularly of the epicotyl hook. The present study aims at clarifying the dynamics of hook formation including the possible involvement of gravity. Time-course studies with normal Alaska pea (Pisum sativum L., cv. Alaska) and an agravitropic pea mutant, ageotropum, under the 1-g conditions and on a 3-D clinostat revealed that (1) the apical hook of the epicotyl forms by the development of the arc-shaped plumule of the embryo existing in the non-germinated seed. The process of formation consists of two stages: development and partial opening, which are controlled by some intrinsic property of the plumule, but not gravity. Approximately when the epicotyl emerges from the seed coat, the hook is established in both pea varieties. In Alaska the established hook is sustained or enhanced by gravity, resulting in a delay of hook opening compared with on a clinostat, which might give an incorrect idea that gravity causes hook formation. (2) During the hook development and opening processes the original plumular arc holds its orientation unchanged to be an established hook, which, therefore, is at the same side of the epicotyl axis as the cotyledons. This is true for both Alaska and ageotropum under 1-g conditions as well as on the clinostat, supporting finding (1). (3) Application of auxin polar transport inhibitors, hydroxyfluorenecarboxylic acid, naphthylphthalamic acid, and triiodobenzoic acid, suppressed the curvature of hook by equal extents in Alaska as well as ageotropum, suggesting that the hook development involves auxin polar transport probably asymmetrically distributed across the plumular axis by some intrinsic property of the plumule not directly related with gravity action. PMID

  11. Analysis of apical hook formation in Alaska pea with a 3-D clinostat and agravitropic mutant ageotropum.

    PubMed

    Miyamoto, Kensuke; Yamasaki, Takahiro; Uheda, Eiji; Ueda, Junichi

    2014-01-01

    The formation of the apical hook in dicotyledonous seedlings is believed to be effected by gravity in the dark. However, this notion is mostly based on experiments with the hook formed on the hypocotyl, and no detailed studies are available with the developmental manners of the hook, particularly of the epicotyl hook. The present study aims at clarifying the dynamics of hook formation including the possible involvement of gravity. Time-course studies with normal Alaska pea (Pisum sativum L., cv. Alaska) and an agravitropic pea mutant, ageotropum, under the 1-g conditions and on a 3-D clinostat revealed that (1) the apical hook of the epicotyl forms by the development of the arc-shaped plumule of the embryo existing in the non-germinated seed. The process of formation consists of two stages: development and partial opening, which are controlled by some intrinsic property of the plumule, but not gravity. Approximately when the epicotyl emerges from the seed coat, the hook is established in both pea varieties. In Alaska the established hook is sustained or enhanced by gravity, resulting in a delay of hook opening compared with on a clinostat, which might give an incorrect idea that gravity causes hook formation. (2) During the hook development and opening processes the original plumular arc holds its orientation unchanged to be an established hook, which, therefore, is at the same side of the epicotyl axis as the cotyledons. This is true for both Alaska and ageotropum under 1-g conditions as well as on the clinostat, supporting finding (1). (3) Application of auxin polar transport inhibitors, hydroxyfluorenecarboxylic acid, naphthylphthalamic acid, and triiodobenzoic acid, suppressed the curvature of hook by equal extents in Alaska as well as ageotropum, suggesting that the hook development involves auxin polar transport probably asymmetrically distributed across the plumular axis by some intrinsic property of the plumule not directly related with gravity action.

  12. Cell length, light and(14)C-labelled indol-3yl-acetic acid transport inPisum satisum L. andPhaseolus vulgaris L.

    PubMed

    Eliezer, J; Morris, D A

    1980-01-01

    The putative auxin-transporting cells of the intact herbaceous dicotyledon are the young, differentiating vascular elements. The length of these cells was found to be considerably greater in dwarf (Meteor) than in tall (Alderman) varieties ofPisum sativum L., and to be greater in etiolated than in light-grown plants ofP. sativum cv Meteor andPhaseolus vulgaris L. cv Mexican Black. Under given light conditions during transport these large differences in cell length did not influence the shapes of the transport profiles or the velocity of transport of(14)C-labelled indol-3yl-acetic acid (IAA) applied to the apical bud. However, in both etiolated and light-grown bean and dwarf pea plants the velocity of transport in darkness was ca. 25% lower than that in light. Under the same conditions of transport velocities in bean were about twice those observed in the dwarf pea. Exposure to light during transport increased the rate of export of(14)C from the labelled shoot apex in green dwarf pea plants but not in etiolated plants. The light conditions to which the plants were exposed during growth and transport had little effect on the rates of uptake of IAA from the applied solutions. The results indicate that the velocity of auxin transport is independent of the frequency of cell-to-cell interfaces along the transport pathway and it is suggested that in intact plants auxin transport is entirely symplastic.

  13. New anthocyanins from purple pods of pea (Pisum spp.).

    PubMed

    Terahara, N; Honda, T; Hayashi, M; Ishimaru, K

    2000-12-01

    Two new anthocyanins were isolated from purple pods of pea (Pisum spp.). Their structures were identified as delphinidin 3-xylosylgalactoside-5-acetylglucoside and its deacetylated derivative by the usual chemical degradation methods and by spectroscopic methods such as UV-VIS, MS and NMR. Both pigments showed moderate stability and antioxidative activity in a neutral aqueous solution.

  14. Pea (Pisum sp.) genetic resources, its analysis and exploration

    USDA-ARS?s Scientific Manuscript database

    Pea is important temperate region pulse, with feed, fodder and vegetable uses. Originated and domesticated in Middle East and Mediterranean, it formed important dietary components of early civilizations. Although Pisum is a small genus with two or three species, it is very diverse and structured, r...

  15. Structure of the coding region and mRNA variants of the apyrase gene from pea (Pisum sativum)

    NASA Technical Reports Server (NTRS)

    Shibata, K.; Abe, S.; Davies, E.

    2001-01-01

    Partial amino acid sequences of a 49 kDa apyrase (ATP diphosphohydrolase, EC 3.6.1.5) from the cytoskeletal fraction of etiolated pea stems were used to derive oligonucleotide DNA primers to generate a cDNA fragment of pea apyrase mRNA by RT-PCR and these primers were used to screen a pea stem cDNA library. Two almost identical cDNAs differing in just 6 nucleotides within the coding regions were found, and these cDNA sequences were used to clone genomic fragments by PCR. Two nearly identical gene fragments containing 8 exons and 7 introns were obtained. One of them (H-type) encoded the mRNA sequence described by Hsieh et al. (1996) (DDBJ/EMBL/GenBank Z32743), while the other (S-type) differed by the same 6 nucleotides as the mRNAs, suggesting that these genes may be alleles. The six nucleotide differences between these two alleles were found solely in the first exon, and these mutation sites had two types of consensus sequences. These mRNAs were found with varying lengths of 3' untranslated regions (3'-UTR). There are some similarities between the 3'-UTR of these mRNAs and those of actin and actin binding proteins in plants. The putative roles of the 3'-UTR and alternative polyadenylation sites are discussed in relation to their possible role in targeting the mRNAs to different subcellular compartments.

  16. Gibberellin (GA3) enhances cell wall invertase activity and mRNA levels in elongating dwarf pea (Pisum sativum) shoots

    NASA Technical Reports Server (NTRS)

    Wu, L. L.; Mitchell, J. P.; Cohn, N. S.; Kaufman, P. B.

    1993-01-01

    The invertase (EC 3.2.1.26) purified from cell walls of dwarf pea stems to homogeneity has a molecular mass of 64 kilodaltons (kD). Poly(A)+RNA was isolated from shoots of dwarf pea plants, and a cDNA library was constructed using lambda gt11 as an expression vector. The expression cDNA library was screened with polyclonal antibodies against pea cell wall invertase. One invertase cDNA clone was characterized as a full-length cDNA with 1,863 base pairs. Compared with other known invertases, one homologous region in the amino acid sequence was found. The conserved motif, Asn-Asp-Pro-Asn-Gly, is located near the N-terminal end of invertase. Northern blot analysis showed that the amount of invertase mRNA (1.86 kb) was rapidly induced to a maximal level 4 h after GA3 treatment, then gradually decreased to the control level. The mRNA level at 4 h in GA3-treated peas was fivefold higher than that of the control group. The maximal increase in activity of pea cell wall invertase elicited by GA3 occcured at 8 h after GA3 treatment. This invertase isoform was shown immunocytochemically to be localized in the cell walls, where a 10-fold higher accumulation occurred in GA3-treated tissue compared with control tissue. This study indicates that the expression of the pea shoot cell-wall invertase gene could be regulated by GA3 at transcriptional and/or translational levels.

  17. Identification and detection of genetic relatedness among important varieties of pea (Pisum sativum L.) grown in India.

    PubMed

    Choudhury, P Ray; Tanveer, Hasan; Dixit, G P

    2007-06-01

    Among the cool season legume crops grown in India and the Indian sub-continent, peas are very popular and preferred by the growers as well as consumers for various uses. The third largest area in pea cultivation is occupied by India after Canada and Russia. Among the important and popular varieties of peas that are grown in India, several are from exotic background. But very little work has been done to carry out the genetic diversity present in the widely adapted Indian pea varieties using DNA markers. Twenty-four most popular and widely adapted varieties were subjected to RAPD analysis to find out the genetic relatedness among them using 60 decamer primers. All the primers used in our study were found to be polymorphic and seven of them showed 100% polymorphism. Out of 579 amplified products, 433 showed polymorphism (74.8%). On an average, 9.65 bands were amplified per primer. Cluster analysis based on Jaccard's similarity coefficient using UPGMA grouped all the tall type varieties together, whereas, dwarf types formed two different clusters based upon their pedigree. The arithmetic mean heterozygosity (Hav) value and marker index (MI) was found to be 0.496 and 4.787, respectively, thus this indicated the efficiency of RAPD as a marker system. Moreover, the calculated value of probability of identical match by chance suggested that about 10(53) genotypes can be unambiguously distinguish by employing 60 RAPD primers.

  18. Comparison of the morphogenesis of three genotypes of pea (Pisum sativum) grown in pure stands and wheat-based intercrops

    PubMed Central

    Barillot, Romain; Combes, Didier; Pineau, Sylvain; Huynh, Pierre; Escobar-Gutiérrez, Abraham J.

    2014-01-01

    Cereal–legume intercrops represent a promising way of combining high productivity and agriculture sustainability. The benefits of cereal–legume mixtures are highly affected by species morphology and functioning, which determine the balance between competition and complementarity for resource acquisition. Studying species morphogenesis, which controls plant architecture, is therefore of major interest. The morphogenesis of cultivated species has been mainly described in mono-specific growing conditions, although morphogenetic plasticity can occur in multi-specific stands. The aim of the present study was therefore to characterize the variability of the morphogenesis of pea plants grown either in pure stands or mixed with wheat. This was achieved through a field experiment that included three pea cultivars with contrasting earliness (hr and HR type) and branching patterns. Results show that most of the assessed parameters of pea morphogenesis (phenology, branching, final number of vegetative organs and their kinetics of appearance) were mainly dependent on the considered genotype, which highlights the importance of the choice of cultivars in intercropping systems. There was however a low variability of pea morphogenesis between sole and mixed stands except for plant height and branching of the long-cycle cultivar. The information provided in the present study at stand and plant scale can be used to build up structural–functional models. These models can contribute to improving the understanding of the functioning of cereal–legume intercrops and also to the definition of plant ideotypes adapted to the growth in intercrops. PMID:24790127

  19. A Comparison of Dryland Grasspea (Lathyrus sativus L.) and Admiral Pea (Pisum sativum L.) Grown Under Different Row Spacings

    USDA-ARS?s Scientific Manuscript database

    Grasspea (GP) is a drought-tolerant legume grown for forage and grain in Europe and the Middle East. It has potential value to be used as a nitrogen-fixing crop in rotation with common grain crops in the High Plains. However, the agronomics of GP for our region have not been investigated to date. W...

  20. Effect of Processing on the in Vitro and in Vivo Protein Quality of Yellow and Green Split Peas (Pisum sativum).

    PubMed

    Nosworthy, Matthew G; Franczyk, Adam J; Medina, Gerardo; Neufeld, Jason; Appah, Paulyn; Utioh, Alphonsus; Frohlich, Peter; House, James D

    2017-09-06

    In order to determine the effect of extrusion, baking, and cooking on the protein quality of yellow and green split peas, a rodent bioassay was conducted and compared to an in vitro method of protein quality determination. The Protein Digestibility-Corrected Amino Acid Score (PDCAAS) of green split peas (71.4%) was higher than that of yellow split peas (67.8%), on average. Similarly, the average Digestible Indispensable Amino Acid Score (DIAAS) of green split peas (69%) was higher than that of yellow split peas (67%). Cooked green pea flour had lower PDCAAS and DIAAS values (69.19% and 67%) than either extruded (73.61%, 70%) or baked (75.22%, 70%). Conversely, cooked yellow split peas had the highest PDCCAS value (69.19%), while extruded yellow split peas had the highest DIAAS value (67%). Interestingly, a strong correlation was found between in vivo and in vitro analysis of protein quality (R(2) = 0.9745). This work highlights the differences between processing methods on pea protein quality and suggests that in vitro measurements of protein digestibility could be used as a surrogate for in vivo analysis.

  1. Structure of the coding region and mRNA variants of the apyrase gene from pea (Pisum sativum)

    NASA Technical Reports Server (NTRS)

    Shibata, K.; Abe, S.; Davies, E.

    2001-01-01

    Partial amino acid sequences of a 49 kDa apyrase (ATP diphosphohydrolase, EC 3.6.1.5) from the cytoskeletal fraction of etiolated pea stems were used to derive oligonucleotide DNA primers to generate a cDNA fragment of pea apyrase mRNA by RT-PCR and these primers were used to screen a pea stem cDNA library. Two almost identical cDNAs differing in just 6 nucleotides within the coding regions were found, and these cDNA sequences were used to clone genomic fragments by PCR. Two nearly identical gene fragments containing 8 exons and 7 introns were obtained. One of them (H-type) encoded the mRNA sequence described by Hsieh et al. (1996) (DDBJ/EMBL/GenBank Z32743), while the other (S-type) differed by the same 6 nucleotides as the mRNAs, suggesting that these genes may be alleles. The six nucleotide differences between these two alleles were found solely in the first exon, and these mutation sites had two types of consensus sequences. These mRNAs were found with varying lengths of 3' untranslated regions (3'-UTR). There are some similarities between the 3'-UTR of these mRNAs and those of actin and actin binding proteins in plants. The putative roles of the 3'-UTR and alternative polyadenylation sites are discussed in relation to their possible role in targeting the mRNAs to different subcellular compartments.

  2. Soil and foliar zinc biofortification in field pea (Pisum sativum L.): Grain accumulation and bioavailability in raw and cooked grains.

    PubMed

    Poblaciones, M J; Rengel, Z

    2016-12-01

    To evaluate the potential of cooked field peas to be used in Zn biofortification programs, all combinations of soil Zn application of 0, 4 and 8mgZnSO4·7H2Okg(-1) and foliar Zn application of 0 and two sprays of 0.25% or 0.5% (w/v) ZnSO4·7H2O before flowering and at early grain-filling stage were tested. Soil Zn application increased Zn-DTPA concentration 3.7- to 5.6-times depending on the Zn soil treatments. Grain Zn concentrations higher than 60mgZnkg(-1) were obtained with all foliar Zn applications, alone or in combination with soil Zn applications, and grain Zn bioavailability was adequate (phytate:Zn ratios lower than 15). Processing (freezing and cooking) caused a decrease of about 30% in grain Zn concentration and a 17%-increase in phytate:Zn ratios (to ⩽9.5). The combined application of 8mgZnSO4·7H2Okg(-1) soil+0.25% (w/v) ZnSO4·7H2O foliarly could be a good option for biofortifying field peas.

  3. Gibberellin (GA3) enhances cell wall invertase activity and mRNA levels in elongating dwarf pea (Pisum sativum) shoots

    NASA Technical Reports Server (NTRS)

    Wu, L. L.; Mitchell, J. P.; Cohn, N. S.; Kaufman, P. B.

    1993-01-01

    The invertase (EC 3.2.1.26) purified from cell walls of dwarf pea stems to homogeneity has a molecular mass of 64 kilodaltons (kD). Poly(A)+RNA was isolated from shoots of dwarf pea plants, and a cDNA library was constructed using lambda gt11 as an expression vector. The expression cDNA library was screened with polyclonal antibodies against pea cell wall invertase. One invertase cDNA clone was characterized as a full-length cDNA with 1,863 base pairs. Compared with other known invertases, one homologous region in the amino acid sequence was found. The conserved motif, Asn-Asp-Pro-Asn-Gly, is located near the N-terminal end of invertase. Northern blot analysis showed that the amount of invertase mRNA (1.86 kb) was rapidly induced to a maximal level 4 h after GA3 treatment, then gradually decreased to the control level. The mRNA level at 4 h in GA3-treated peas was fivefold higher than that of the control group. The maximal increase in activity of pea cell wall invertase elicited by GA3 occcured at 8 h after GA3 treatment. This invertase isoform was shown immunocytochemically to be localized in the cell walls, where a 10-fold higher accumulation occurred in GA3-treated tissue compared with control tissue. This study indicates that the expression of the pea shoot cell-wall invertase gene could be regulated by GA3 at transcriptional and/or translational levels.

  4. The influence of dietary field peas (Pisum sativum L.) on pig performance, carcass quality, and the palatability of pork.

    PubMed

    Stein, H H; Everts, A K R; Sweeter, K K; Peters, D N; Maddock, R J; Wulf, D M; Pedersen, C

    2006-11-01

    An experiment was conducted to test the hypothesis that field peas may replace soybean meal in diets fed to growing and finishing pigs without negatively influencing pig performance, carcass quality, or pork palatability. Forty-eight pigs (initial average BW 22.7 +/- 1.21 kg) were allotted to 1 of 3 treatments with 2 pigs per pen. There were 8 replications per treatment, 4 with barrows and 4 with gilts. The treatments were control, medium field peas, and maximum field peas. Pigs were fed grower diets for 35 d, early finisher diets for 35 d, and late finisher diets for 45 d. Pigs receiving the control treatment were fed corn-soybean meal diets. All diets fed to pigs receiving the medium field peas treatment contained 36% field peas and varying amounts of corn; soybean meal was also included in the grower and the early finisher diets fed to pigs on this treatment. In contrast, no soybean meal was included in diets fed to pigs on the maximum field peas treatment, and field peas were included at concentrations of 66, 48, and 36% in the grower, early finisher, and late finisher diets, respectively. Pig performance was monitored within each phase and for the entire experimental period. At the conclusion of the experiment, carcass composition, carcass quality, and the palatability of pork chops and pork patties were measured. Results showed that there were no effects of dietary treatments on ADFI, ADG, or G:F. Likewise, there were no differences in carcass composition among the treatment groups, but gilts had larger (P = 0.001) and deeper (P = 0.003) LM, less backfat (P = 0.007), and a greater (P = 0.002) lean meat percentage than barrows. The pH and marbling of the LM, and the 10th rib backfat were not influenced by treatment, but there was a trend (P = 0.10) for more marbling in barrows than in gilts. The subjective color scores (P = 0.003) and the objective color score (P = 0.06) indicated that dietary field peas made the LM darker and more desirable. Pork chops from pigs fed field peas also had less (P = 0.02) moisture loss compared with chops from pigs fed the control diet. Treatment or sex did not influence palatability of pork chops or pork patties. In conclusion, field peas may replace all of the soybean meal in diets fed to growing and finishing pigs without negatively influencing pig performance, carcass composition, carcass quality, or pork palatability.

  5. Host Adaptation of Soybean Dwarf Virus Following Serial Passages on Pea (Pisum sativum) and Soybean (Glycine max).

    PubMed

    Tian, Bin; Gildow, Frederick E; Stone, Andrew L; Sherman, Diana J; Damsteegt, Vernon D; Schneider, William L

    2017-06-21

    Soybean Dwarf Virus (SbDV) is an important plant pathogen, causing economic losses in soybean. In North America, indigenous strains of SbDV mainly infect clover, with occasional outbreaks in soybean. To evaluate the risk of a US clover strain of SbDV adapting to other plant hosts, the clover isolate SbDV-MD6 was serially transmitted to pea and soybean by aphid vectors. Sequence analysis of SbDV-MD6 from pea and soybean passages identified 11 non-synonymous mutations in soybean, and six mutations in pea. Increasing virus titers with each sequential transmission indicated that SbDV-MD6 was able to adapt to the plant host. However, aphid transmission efficiency on soybean decreased until the virus was no longer transmissible. Our results clearly demonstrated that the clover strain of SbDV-MD6 is able to adapt to soybean crops. However, mutations that improve replication and/or movement may have trade-off effects resulting in decreased vector transmission.

  6. Foliar application of methyl jasmonate induced physio-hormonal changes in Pisum sativum under diverse temperature regimes.

    PubMed

    Shahzad, Raheem; Waqas, Muhammad; Khan, Abdul Latif; Hamayun, Muhammad; Kang, Sang-Mo; Lee, In-Jung

    2015-11-01

    Global climate change brings with it unwarranted shifts in both abiotic (heat stress, cold stress, wind, precipitation) and biotic (pathogens, pests) environmental factors, thus posing a threat to agricultural productivity across the world. In plants, lodging due to storms or herbivory causes wounding stress and consequently enhances endogenous jasmonates. In response, the plant growth is arrested as plant defense is prioritized. We pre-treated pea plants with elevated methyl jasmonate (MeJA) levels i.e. 50 μM, 100 μM and 200 μM under controlled growth chamber conditions. The pre-treated plants were then kept at 40 °C (heat stress--HS), 4 °C (cold stress--CS) and 20 °C (optimum/control temperature--OT) for 72 h. The effect of such treatments on plant growth attributes, photosynthesis, stomatal conductance, cell death rate, and regulation of endogenous hormones were observed. Elevated MeJA application hindered plant growth attributes under HS, CS and OT conditions. Moreover, elevated MeJA levels lowered the rate of photosynthesis and stomatal conductance, induced stomatal closure, caused higher cells mortality in leaves under HS, CS, and OT conditions. Endogenous ABA contents significantly declined in all MeJA treatments under HS and OT, but increased under CS conditions. Exogenous MeJA enhanced endogenous jasmonic acid contents of pea plants, but altered endogenous salicylic acid contents under varying temperatures. Current study shows that higher concentrations of exogenous MeJA strengthen plant defense mechanism by hindering plant growth under stress conditions.

  7. Water deficit down-regulates miR398 and miR408 in pea (Pisum sativum L.).

    PubMed

    Jovanović, Živko; Stanisavljević, Nemanja; Mikić, Aleksandar; Radović, Svetlana; Maksimović, Vesna

    2014-10-01

    MicroRNAs (miRNAs), recently recognized as important regulator of gene expression at posttranscriptional level, have been found to be involved in plant stress responses. The observation that some miRNAs are up- or down regulated by stress implies that they could play vital roles in plant resistance to abiotic and biotic stress. We investigated the effect of water stress treatment during 10 days on expression of conserved miRNAs-miR398a/b and miR408 in pea plants. This time frame reflects the changes as close as possible to the changes where water stress causes visible effects under field condition. It was observed that dehydration strongly down regulates the expression of both miR398a/b and miR408 in pea roots and shoots. The down-regulation of miR398a/b and the up-regulation of potential target genes - copper superoxide dismutase, CSD1, highlight the involvement of this miRNA in pea stress response. To the contrary, the mRNA level of cytochrome c oxidase subunit 5 (COX5b) did not change in roots and shoots of water-stressed plants, compared to control (well) hydrated plants. This suggests that COX5b is not the target of miR398, or that its expression is regulated by some other mechanism. P1B-ATPase expression increased during water deficit only in the shoots of pea; in the roots there were no changes in expression. Our results help to understand the possible role of investigated miRNAs and their contribution to pea capacity to cope with water deficit. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  8. A Single, Plastic Population of Mycosphaerella pinodes Causes Ascochyta Blight on Winter and Spring Peas (Pisum sativum) in France

    PubMed Central

    Guibert, Michèle; Leclerc, Aurélie; Andrivon, Didier; Tivoli, Bernard

    2012-01-01

    Plant diseases are caused by pathogen populations continuously subjected to evolutionary forces (genetic flow, selection, and recombination). Ascochyta blight, caused by Mycosphaerella pinodes, is one of the most damaging necrotrophic pathogens of field peas worldwide. In France, both winter and spring peas are cultivated. Although these crops overlap by about 4 months (March to June), primary Ascochyta blight infections are not synchronous on the two crops. This suggests that the disease could be due to two different M. pinodes populations, specialized on either winter or spring pea. To test this hypothesis, 144 pathogen isolates were collected in the field during the winter and spring growing seasons in Rennes (western France), and all the isolates were genotyped using amplified fragment length polymorphism (AFLP) markers. Furthermore, the pathogenicities of 33 isolates randomly chosen within the collection were tested on four pea genotypes (2 winter and 2 spring types) grown under three climatic regimes, simulating winter, late winter, and spring conditions. M. pinodes isolates from winter and spring peas were genetically polymorphic but not differentiated according to the type of cultivars. Isolates from winter pea were more pathogenic than isolates from spring pea on hosts raised under winter conditions, while isolates from spring pea were more pathogenic than those from winter pea on plants raised under spring conditions. These results show that disease developed on winter and spring peas was initiated by a single population of M. pinodes whose pathogenicity is a plastic trait modulated by the physiological status of the host plant. PMID:23023742

  9. A single, plastic population of Mycosphaerella pinodes causes ascochyta blight on winter and spring peas (Pisum sativum) in France.

    PubMed

    Le May, Christophe; Guibert, Michèle; Leclerc, Aurélie; Andri