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Are Chemical Compounds Important for Soybean Resistance to Anticarsia gemmatalis ?  

Microsoft Academic Search

The identification and quantification of flavonoids (rutin and genistin) present in extracts of soybean genotypes, and their effects on the biology and physiology of Anticarsia gemmatalis Hübner (Lep.: Noctuidae) were studied. Analysis of covariance and bicoordinate utilization plots were used to remove the effect of feeding time from pupal weight and consumption as well as to separate pre- and postingestive

Giorla Carla Piubelli; Clara Beatriz Hoffmann-Campo; Flávio Moscardi; Sandra Hitomi Miyakubo; Maria Cristina Neves De Oliveira



Are chemical compounds important for soybean resistance to Anticarsia gemmatalis?  


The identification and quantification of flavonoids (rutin and genistin) present in extracts of soybean genotypes, and their effects on the biology and physiology of Anticarsia gemmatalis Hübner (Lep.: Noctuidae) were studied. Analysis of covariance and bicoordinate utilization plots were used to remove the effect of feeding time from pupal weight and consumption as well as to separate pre- and postingestive effects of treatment on A. gemmatalis growth. Genotypes PI 274454, PI 227687, and "IAC-100" extracts in general, caused higher mortality, negatively influenced initial larval and pupal weight, and elongated larval cycle. Larvae fed on the "IAC-100" extract diet ingested larger amounts of food per unit of time, but were less efficient in its conversion to biomass. Leaf extracts of PI 227687 had the largest concentration of rutin (quercitin 3-O-rhamnosylglucoside), followed by PI 274454, and "IAC-100"; PI 74454 also had the highest genistin (genistein 7-O-glucoside) content. The susceptible cultivar "BR-16" showed only a kaempferol-based flavonoid in its chemical profile, indicating that after successive crosses, secondary compounds responsible for plant defenses were eliminated. Genotypes PI 274454, PI 227687, and "IAC-100" showed accentuated resistance characteristics and were considered inadequate sources for the development of A. gemmatalis. Considering rutin and genistin concentration in these genotypes, it is suggested that flavonoids are important factors conferring resistance to A. gemmatalis. PMID:16222789

Piubelli, Giorla Carla; Hoffmann-Campo, Clara Beatriz; Moscardi, Flavio; Miyakubo, Sandra Hitomi; de Oliveira, Maria Cristina Neves



Physical maps and virulence of Anticarsia gemmatalis nucleopolyhedrovirus genomic variants  

Microsoft Academic Search

Summary.  ?Seventeen plaque purified isolates of two viral preparations of Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV), were analyzed in terms of the genomic changes after digestion of their DNAs with\\u000a HindIII and PstI restriction enzymes. The 1979 AgMNPV wild type preparation (AgMNPV-’79) resulted in six different variants\\u000a and the 1985 viral commercial preparation (AgMNPV-’85), in eleven. The genomic variation of all the

J. E. Maruniak; A. Garcia-Maruniak; M. L. Souza; P. M. A. Zanotto; F. Moscardi



Microsomal oxidation of allelochemicals in generalist ( Spodoptera frugiperda ) and semispecialist ( Anticarsia gemmatalis ) insect  

Microsoft Academic Search

Midgut microsomes prepared from larvae of the fall armyworm [Spodoptera frugiperda (J.E Smith)], a generalist insect, and the velvetbean caterpillar (Anticarsia gemmatalis Hubner), a semispecialist, were used to study their oxidative activity toward a variety of allelochemicals. Allelochemicals such as terpenoids, alkaloids, indoles, glucosinolates, flavonoids, coumarins, cardenolides, phenylpropenes, and a ketohydrocarbon were all metabolized by the microsomal cytochrome P-450 monooxygenases

S. J. Yu



Detection of protease inhibitors in the hemolymph of resistant Anticarsia gemmatalis which are inhibitory to the entomopathogenic fungus, Nomuraea rileyi  

Microsoft Academic Search

Summary A complex of protease inhibitor activities has been detected in the hemolymph of the 6th instarAnticarsia gemmatalis larvae that are resistant to infection by the fungusNomuraea rileyi. A site-specific serine protease inhibitor extracted fromA. gemmatalis hemolymph inhibits both the germination ofN. rileyi conidia and subsequent germ tube development.

D. G. Boucias; J. C. Pendland



Classical biological control in an ephemeral crop habitat with Anticarsia gemmatalis nucleopolyhedrovirus  

Microsoft Academic Search

Anticarsia gemmatalis nucleopolyhedrovirus(AgNPV) was released as a single spray in soybean at two sites in Louisiana, near Crowley and Baton Rouge,\\u000a after which viral prevalence and population density were monitored for 3--4 years. Each site had a plot with no treatment\\u000a (control) and two virus-treated plots, one planted with soybean yearly for 3-4 years, and the other planted with soybean

James R. Fuxa; Arthur R. Richter



Pathogenesis of Autographa californica multiple nucleopolyhedrovirus in fifth-instar Anticarsia gemmatalis larvae.  


We have investigated infection and pathogenesis of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in Anticarsia gemmatalis (velvetbean caterpillar) larvae using a lacZ recombinant virus (AcMNPV-hsp70/lacZ) to track the temporal progression of infection in the midgut intestine and haemocoel. A. gemmatalis was highly resistant to fatal infection by occlusion bodies (OBs; LD(50)>5.5 x 10(5) OB) and budded virus (BV; LD(50)>3 x 10(5) BV) administered via oral and systemic routes, respectively. Orally administered occlusion-derived virus (ODV) efficiently attached and fused to midgut cells; however, high levels of infection-induced apoptosis limited infection in the midgut. Transcriptional analysis of AcMNPV genes expressed in the midgut of OB-inoculated A. gemmatalis larvae showed high levels of mRNA encoding the major capsid protein VP39 in the absence of immediate-early transactivator 1 (ie-1) expression. In the midgut, virus was efficiently transferred from infected midgut epithelial cells to nearby tracheolar cells and circulating haemocytes to initiate systemic infection in the haemocoel. However, haemocoelic BV did not efficiently disseminate infection and only cuticular epidermal cells displayed high levels of viral infection. Flow cytometry analysis of haemocytes isolated from BV-inoculated A. gemmatalis larvae showed low-level expression of the BV envelope protein GP64 on the cell surface, suggesting that A. gemmatalis haemocytes have a limited capacity for amplifying virus. These results show that AcMNPV is not an effective biological control agent for limiting crop damage caused by A. gemmatalis larvae. PMID:19423548

Chikhalya, Aniska; Luu, Dee Dee; Carrera, Maggie; De La Cruz, Alisa; Torres, Marianne; Martinez, Elisa N; Chen, Tiffany; Stephens, Kimberly D; Haas-Stapleton, Eric J



Hemagglutinin activity in the hemolymph of Anticarsia gemmatalis larvae infected with the fungus Nomuraea rileyi.  


Hemolymph samples from larvae of 3 lepidopteran species (Anticarsia gemmatalis, Trichoplusia ni and Spodoptera frugiperda) were tested for hemagglutination activity. Samples from A. gemmatalis larvae which had been injected 12-24 hrs previously with hyphal bodies of the entomopathogenic fungus Nomuraea rileyi showed agglutination activity against human 0, rabbit and sheep erythrocytes. Little or no activity was detected in samples from the other 2 larval species. Low titers (approximately 1:2) were observed when rabbit and sheep erythrocytes were tested with hemolymph from non-injected or water-injected A. gemmatalis control larvae. Higher titers (1:256-1:1024) were obtained when human erythrocytes were incubated with control hemolymph, but values were greater in the hyphal body-injected samples (1:2048-1: greater than 32,000). These results indicate a direct correlation between agglutinin production and the presence of fungal cells in the larval hemolymph. Injection with heat-killed or homogenated hyphal bodies did not cause increased activity. Decreases in titer values after these injections and for 1-12 hrs after injection with viable hyphal bodies suggest that the agglutinin(s) may function in immune surveillance. Agglutination of rabbit erythrocytes was inhibited by lactose, galactose and L-fucose. N-acetylneuraminic acid inhibited agglutination of human erythrocytes. PMID:3996705

Pendland, J C; Boucias, D G



Effects of Fetal Bovine Serum deprivation in cell cultures on the production of Anticarsia gemmatalis Multinucleopolyhedrovirus  

PubMed Central

Background Anticarsia gemmatalis is a pest in South America's soybean crops, which could be controlled by the Multinucleopolyhedrovirus of A. gemmatalis (AgMNPV). Currently, its commercial production is based on infected larvae. However, the possibility of using modified baculoviruses in Integrated Pest Management programs has stimulated an interest to develop alternative multiplication processes. This study evaluated the AgMNPV production in UFL-Ag-286 cells previously deprived Fetal Bovine Serum. Results Culture media containing 1% FBS during the previous 48 hours achieved a synchronized condition where 90% of cells were found in G0/G1 stage, showing the presence of non-filamentous actin. All characteristics were estimated from cellular viability tests, cell actin detection trials and flow cytometer cell cycle analysis. AgMNPV production was tested by transcript studies and budded viruses (BVs) and occlusion bodies (OBs) yield quantitation. Results showed that the productivity in FBS deprived cells was 9.8 times more in BVs and 3.8 times more in OBs with respect to non-treated cells. Conclusions UFL-Ag-286 cells previously deprived in FBS shown to be a better host for AgMNPV propagation, increasing the useful for both in vitro bioinsecticide production and applications such as recombinant protein expression or gene delivery.



The larval midgut of Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae): light and electron microscopy studies of the epithelial cells.  


The morphology of the midgut epithelium cells of Anticarsia gemmatalis (Hübner) larvae is described by light and transmission electron microscopy. The midgut of A. gemmatalis is the largest portion of the digestive tract, with three distinct regions: proximal, media and distal. Its wall is formed by pseudostratified columnar epithelial tissue having four cell types: columnar, goblet, regenerative, and endocrine cells. The columnar cells are numerous and long, with the apical portion showing many lengthy microvilli and the basal portion invaginations forming a basal labyrinth. The goblet cells have a large goblet-shaped central cavity delimited by cytoplasmic projections filled with mitochondria. The regenerative cells present electron-dense cytoplasm and few organelles. The endocrine cells are characterized by electron-dense secretory granules, usually concentrated in the cytoplasm basal region. PMID:15620001

Levy, S M; Falleiros, A M F; Gregório, E A; Arrebola, N R; Toledo, L A



Proteolytic activity of gut bacteria isolated from the velvet bean caterpillar Anticarsia gemmatalis.  


The development of proteinase inhibitors as potential insect control agents has been constrained by insect adaptation to these compounds. The velvet bean caterpillar (Anticarsia gemmatalis) is a key soybean pest species that is well-adapted to proteinase inhibitors, particularly serine-proteinase inhibitors, which are abundant in the caterpillar host. The expression of diverse proteolytic enzymes by gut symbionts may allow the velvet bean caterpillar to circumvent proteinase inhibitors produced by the host plant. In this study, we characterized the proteolytic activity of the four nonpathogenic species of gut bacteria isolated from the velvet bean caterpillar-Bacillus cereus, Enterococcus gallinarum, Enterococcus mundtii and Staphylococcus xylosus. Two proteinase substrates, N-?-benzoyl-L-Arg-p-nitroanilide (L-BApNA) and N-?-p-tosyl-L-Arg methyl ester (L-TAME) and five proteinase inhibitors [aprotinin, E-64, ethylenediamine tetraacetic acid (EDTA), pepstatin and N-?-tosyl-L-lysine chloromethyl ketone (TLCK)] as well as CaCl2, pH and temperature profiles were used to characterize the expressed proteolytic activity of these bacterial strains in vitro. Kinetic parameters for proteolytic activity were also estimated. The results of these experiments indicated that serine- and cysteine-proteinase activities were expressed by all four gut bacteria symbionts of the velvet bean caterpillar. The cysteine- and serine-proteinase activities of these gut symbionts were distinct and different from that of gut proteinases of the caterpillar itself. This finding provides support for the potential involvement of gut symbionts in the mitigation of the negative effects of serine-proteinase inhibitors in the velvet bean caterpillar. PMID:23392900

Pilon, F M; Visôtto, L E; Guedes, R N C; Oliveira, M G A



Do different casein concentrations increase the adverse effect of rutin on the biology of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae)?  


The flavonoid rutin is recognized as playing an important role in the protection of plants against lepidopterans. Bioassays with this compound are generally carried out using artificial diets. Proteins of high energy value, such as casein, are important ingredients of insect artificial diets as a source of essential amino acids. However, such proteins can generally increase the allelochemical activity. Our objective was to evaluate the effects of rutin on larvae of the velvetbean caterpillar Anticarsia gemmatalis Hübner by incorporating this allelochemical into diets with different concentrations of casein. Three casein concentrations (0, 7 g, or 14 g) combined with none, 0.65%, or 1.30% of rutin were added to the rearing diet and offered to the larvae from hatching to pupation. Rutin negatively affected larval development, the amount of food consumed, and pupal weight of A. gemmatalis. These negative effects were clearly seen in insects fed on diets with 7 g of casein to which any concentration of rutin was added. The effects of rutin when added to the diets without casein were stronger than in diets containing a suitable amount of casein (14 g). The greater negative effects of rutin in diets containing suboptimal concentrations of casein indicate that casein can increase the effects of rutin only when the diets are nutritionally unsuitable for insect development. PMID:21120388

Salvador, Mariana C; Boiça, Arlindo L; de Oliveira, Maria C N; da Graça, José P; da Silva, Débora M; Hoffmann-Campo, Clara B


Inactivation of the ecdysteroid UDP-glucosyltransferase ( egt) gene of Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) improves its virulence towards its insect host  

Microsoft Academic Search

Some baculovirus have been genetically modified for the inactivation of their ecdysteroid glucosyltransferase (egt) gene, and these viruses were shown to kill infected larvae more rapidly when compared to wild-type virus infections. We have previously identified, cloned, and sequenced the egt gene of Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV). Here we present data regarding the construction of an egt minus (egt?) AgMNPV

Francisco J. R. Pinedo; Flávio Moscardi; Teresa Luque; Julie A. Olszewski; Bergmann M. Ribeiro



Screening of Brazilian Bacillus thuringiensis isolates active against Spodoptera frugiperda, Plutella xylostella and Anticarsia gemmatalis  

Microsoft Academic Search

The toxicity of a collection of 1400 isolates of Bacillus thuringiensis was assessed against the Lepidoptera Spodoptera frugiperda, Anticarsia gemmantalis and Plutella xylostella. Twenty seven isolates showed toxicity to the larvae of these insects with three isolates demonstrating significantly greater potency than the standard strain against Lepidoptera, B. thuringiensis serovar kurstaki HD1. These isolates were all found to produce bipyramidal

Rose Gomes Monnerat; Andrea Cardoso Batista; Patrícia Telles de Medeiros; Érica Soares Martins; Viviane M. Melatti; Lílian Botelho Praça; Vinicius Fiúza Dumas; Cristiane Morinaga; Caroline Demo; Ana Cristina Menezes Gomes; Rosana Falcão; Claudia Brod Siqueira; Joseilde Oliveira Silva-Werneck; Colin Berry



A Recombinant Anticarsia gemmatalis MNPV Harboring chiA and v-cath Genes from Choristoneura fumiferana Defective NPV Induce Host Liquefaction and Increased Insecticidal Activity.  


One of the interesting features of Anticarsia gemmatalis multiple nucleopolyhedrovirus isolate 2D (AgMNPV-2D) genome is the absence of chitinase (chiA) and cathepsin (v-cath) genes. This characteristic may be responsible for the lack of liquefaction and melanization in A. gemmatalis larvae killed by AgMNPV-2D infection. This study aimed to test the hypothesis that CHIA and V-CATH proteins from Choristonera fumiferana DEF multiple nucleopolyhedrovirus (CfDEFNPV) are able to liquefy and melanize the cuticle of A. gemmatalis larvae infected by a recombinant AgMNPV containing chiA and v-cath genes inserted in its genome. A fragment from the CfDefNPV genome containing chiA and v-cath genes was inserted into the genome of AgMNPV-2D. The recombinant virus (vAgp2100Cf.chiA/v-cath) was purified and used to infect insect cells and larvae. Transcripts of v-cath and chiA genes were detected along the infection of insect cells by qRT-PCR, from early to late phases of infection. The analysis of A. gemmatalis larvae killed by vAgp2100Cf.chiA/v-cath infection confirmed the hypothesis proposed. The vAgp2100Cf.chiA/v-cath showed higher insecticidal activity against third instar A. gemmatalis larvae when compared to AgMNPV-2D. The mean time to death was also lower for the vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D at 10 days post infection. Occlusion body production was higher in A. gemmatalis larvae infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. Enzyme assays showed higher chitinase and cysteine protease activities in insect cells and insects infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. The introduction of chiA and v-cath genes into the genome of AgMNPV improves its insecticidal activity against A. gemmatalis larvae and this recombinant virus could be used as an alternative to the wild type virus to control this important insect pest. PMID:24086357

Lima, Anabele Azevedo; Aragão, Clara Wandenkolck Silva; de Castro, Maria Elita Batista; Oliveira, Juliana Velasco de Castro; Sosa Gómez, Daniel Ricardo; Ribeiro, Bergmann Morais



A Recombinant Anticarsia gemmatalis MNPV Harboring chiA and v-cath Genes from Choristoneura fumiferana Defective NPV Induce Host Liquefaction and Increased Insecticidal Activity  

PubMed Central

One of the interesting features of Anticarsia gemmatalis multiple nucleopolyhedrovirus isolate 2D (AgMNPV-2D) genome is the absence of chitinase (chiA) and cathepsin (v-cath) genes. This characteristic may be responsible for the lack of liquefaction and melanization in A. gemmatalis larvae killed by AgMNPV-2D infection. This study aimed to test the hypothesis that CHIA and V-CATH proteins from Choristonera fumiferana DEF multiple nucleopolyhedrovirus (CfDEFNPV) are able to liquefy and melanize the cuticle of A. gemmatalis larvae infected by a recombinant AgMNPV containing chiA and v-cath genes inserted in its genome. A fragment from the CfDefNPV genome containing chiA and v-cath genes was inserted into the genome of AgMNPV-2D. The recombinant virus (vAgp2100Cf.chiA/v-cath) was purified and used to infect insect cells and larvae. Transcripts of v-cath and chiA genes were detected along the infection of insect cells by qRT-PCR, from early to late phases of infection. The analysis of A. gemmatalis larvae killed by vAgp2100Cf.chiA/v-cath infection confirmed the hypothesis proposed. The vAgp2100Cf.chiA/v-cath showed higher insecticidal activity against third instar A. gemmatalis larvae when compared to AgMNPV-2D. The mean time to death was also lower for the vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D at 10 days post infection. Occlusion body production was higher in A. gemmatalis larvae infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. Enzyme assays showed higher chitinase and cysteine protease activities in insect cells and insects infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. The introduction of chiA and v-cath genes into the genome of AgMNPV improves its insecticidal activity against A. gemmatalis larvae and this recombinant virus could be used as an alternative to the wild type virus to control this important insect pest.

Lima, Anabele Azevedo; Aragao, Clara Wandenkolck Silva; de Castro, Maria Elita Batista; Oliveira, Juliana Velasco de Castro; Sosa Gomez, Daniel Ricardo; Ribeiro, Bergmann Morais



Molecular analysis of a mutant Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) shows an interruption of an inhibitor of apoptosis gene (iap-3) by a new class-II piggyBac-related insect transposon.  


A new piggyBac-related transposable element (TE) was found in the genome of a mutant Anticarsia gemmatalis multiple nucleopolyhedrovirus interrupting an inhibitor of apoptosis gene. This mutant virus induces apoptosis upon infection of an Anticarsia gemmatalis cell line, but not in a Trichoplusia ni cell line. The sequence of the new TE (which was named IDT for iap disruptor transposon) has 2531 bp with two DNA sequences flanking a putative Transposase (Tpase) ORF of 1719 bp coding for a protein with 572 amino acids. These structural features are similar to the piggyBac TE, also reported for the first time in the genome of a baculovirus. We have also isolated variants of this new TE from different lepidopteran insect cells and compared their Tpase sequences. PMID:19788700

Carpes, M P; Nunes, J F; Sampaio, T L; Castro, M E B; Zanotto, P M A; Ribeiro, B M



Development of Resistance by Anticarsia gemmatalisfrom Brazil and the United States to a Nuclear Polyhedrosis Virus under Laboratory Selection Pressure  

Microsoft Academic Search

Colonies of velvetbean caterpillar,Anticarsia gemmatalis,collected in Brazil (Dourados, State of Mato Grosso do Sul; and Sertanopolis, State of Paraná) and in the United States (Louisiana) were established in the laboratory, in each respective country, with the objective of evaluating the potential of this insect for developing resistance to theA. gemmatalisnuclear polyhedrosis virus (AgMNPV). Larvae of the three colonies were exposed

A. R. Abot; F. Moscardi; J. R. Fuxa; D. R. Sosa-Gómez; A. R. Richter



Structural and ultrastructural alterations of Malpighian tubules of Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae) larvae infected with different Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) recombinant viruses.  


Malpighian tubules constitute the main excretion organ of insects. Infection by egt(-) recombinant AcMNPV baculovirus in lepidopteran larvae promotes early degeneration of these structures, which has been correlated with earlier death of the host. However, no trace of viral infection has been detected in that tissue. We constructed two AgMNPV recombinants with the egfp gene under control of the hsp70 promoter, one being egt(-), and used another two recombinants (one egt(-)) containing the lacZ gene. Morphological alterations in the tubules were analyzed by light and electron microscopies. Bioassays were conducted to compare the pathogenicity of recombinants. Results showed progressive presence of marker proteins and tissue degeneration without signals of infection in the tissue. Morphological and bioassay results showed increased pathogenicity for lacZ-containing recombinants compared to the egfp ones; as for egt(-) viruses, we noted higher intensity and earlier onset of alterations. The absence of infection led us to believe that Malpighian tubules degeneration is provoked initially by the death of tracheal cells attached to the tubules and later, by the death of Malpighian tubule cells themselves. Tubule cell death might be due to oncosis and apoptosis, which may be activated by depletion of energy reserves and by accumulation of marker proteins, respectively. Absence of the egt gene may be leading to a higher energetic expense due to molting, thus aggravating tubule cell death, resulting in faster death of host. PMID:18304572

Cordeiro, Bruno A; Tibúrcio, Victor Hugo S; Hallwass, Mariana; Paes, Hugo C; Ribeiro, Bergmann M; Báo, Sônia N



Contribution of gut bacteria to digestion and development of the velvetbean caterpillar, Anticarsia gemmatalis  

Microsoft Academic Search

Bacteria colonies from gut homogenates of fifth instar velvetbean caterpillars (Lepidoptera: Noctuidae) were subjected to antibiotic sensitivity experiments using discs containing 22 antibiotics. The antibiotic tetracycline provided the best results, followed by chloramphenicol. Tetracycline also provided higher inhibition of colony forming units than chloramphenicol and was therefore provided to the caterpillars in increasing diet concentrations to assess the contribution of

L. E. Visôtto; M. G. A. Oliveira; R. N. C. Guedes; A. O. B. Ribon; P. I. V. Good-God



Interspecific hybrids between the homothallic Phytophthora sojae and Phytophthora vignae  

Microsoft Academic Search

An interspecific cross was attempted between two homothallic species of Phytophthora, P. sojae and P. vignae. From 1640 single-oospore cultures isolated, DNA was extracted from 800, and two interspecific F1 hybrids (F1 1121 and F1 1426) were putatively identified using RAPD markers. The true hybrid nature of these F1 hybrids was confirmed using additional AFLP analysis. Single-zoospore cultures were generated

K. J. May; A. Drenth; J. A. G. Irwin



The genomic relationship between Glycine max (L.) Merr. and G. soja Sieb. and Zucc. as revealed by pachytene chromosome analysis  

Microsoft Academic Search

This study was conducted with the objective of determining the genomic relationship between cultivated soybean (Glycine max) and wild soybean (G. soja) of the subgenus Soja, genus Glycine. Observations on cross-ability rate, hybrid viability, meiotic chromosome pairing, and pollen fertility in F1 hybrids of G. max × G. soja and reciprocals elucidated that both species hybridized readily and set mature

R. J. Singh; T. Hymowitz



Comparative Analysis of Expressed Sequences in Phytophthora sojae1  

PubMed Central

Phytophthora sojae (Kaufmann and Gerdemann) is an oomycete that causes stem and root rot on soybean (Glycine max L. Merr) plants. We have constructed three cDNA libraries using mRNA isolated from axenically grown mycelium and zoospores and from tissue isolated from plant hypocotyls 48 h after inoculation with zoospores. A total of 3,035 expressed sequence tags (ESTs) were generated from the three cDNA libraries, representing an estimated 2,189 cDNA transcripts. The ESTs were classified according to putative function based on similarity to known proteins, and were analyzed for redundancy within and among the three source libraries. Distinct expression patterns were observed for each library. By analysis of the percentage G+C content of the ESTs, we estimate that two-thirds of the ESTs from the infected plant library are derived from P. sojae cDNA transcripts. The ESTs originating from this study were also compared with a collection of Phytophthora infestans ESTs and with all other non-human ESTs to assess the similarity of the P. sojae sequences to existing EST data. This collection of cDNA libraries, ESTs, and accompanying annotation will provide a new resource for studies on oomycetes and on soybean responses to pathogen challenge.

Qutob, Dinah; Hraber, Peter T.; Sobral, Bruno W.S.; Gijzen, Mark



Draft genome sequencing and comparative analysis of Aspergillus sojae NBRC4239.  


We conducted genome sequencing of the filamentous fungus Aspergillus sojae NBRC4239 isolated from the koji used to prepare Japanese soy sauce. We used the 454 pyrosequencing technology and investigated the genome with respect to enzymes and secondary metabolites in comparison with other Aspergilli sequenced. Assembly of 454 reads generated a non-redundant sequence of 39.5-Mb possessing 13 033 putative genes and 65 scaffolds composed of 557 contigs. Of the 2847 open reading frames with Pfam domain scores of >150 found in A. sojae NBRC4239, 81.7% had a high degree of similarity with the genes of A. oryzae. Comparative analysis identified serine carboxypeptidase and aspartic protease genes unique to A. sojae NBRC4239. While A. oryzae possessed three copies of ?-amyalse gene, A. sojae NBRC4239 possessed only a single copy. Comparison of 56 gene clusters for secondary metabolites between A. sojae NBRC4239 and A. oryzae revealed that 24 clusters were conserved, whereas 32 clusters differed between them that included a deletion of 18 508 bp containing mfs1, mao1, dmaT, and pks-nrps for the cyclopiazonic acid (CPA) biosynthesis, explaining the no productivity of CPA in A. sojae. The A. sojae NBRC4239 genome data will be useful to characterize functional features of the koji moulds used in Japanese industries. PMID:21659486

Sato, Atsushi; Oshima, Kenshiro; Noguchi, Hideki; Ogawa, Masahiro; Takahashi, Tadashi; Oguma, Tetsuya; Koyama, Yasuji; Itoh, Takehiko; Hattori, Masahira; Hanya, Yoshiki



Draft Genome Sequencing and Comparative Analysis of Aspergillus sojae NBRC4239  

PubMed Central

We conducted genome sequencing of the filamentous fungus Aspergillus sojae NBRC4239 isolated from the koji used to prepare Japanese soy sauce. We used the 454 pyrosequencing technology and investigated the genome with respect to enzymes and secondary metabolites in comparison with other Aspergilli sequenced. Assembly of 454 reads generated a non-redundant sequence of 39.5-Mb possessing 13 033 putative genes and 65 scaffolds composed of 557 contigs. Of the 2847 open reading frames with Pfam domain scores of >150 found in A. sojae NBRC4239, 81.7% had a high degree of similarity with the genes of A. oryzae. Comparative analysis identified serine carboxypeptidase and aspartic protease genes unique to A. sojae NBRC4239. While A. oryzae possessed three copies of ?-amyalse gene, A. sojae NBRC4239 possessed only a single copy. Comparison of 56 gene clusters for secondary metabolites between A. sojae NBRC4239 and A. oryzae revealed that 24 clusters were conserved, whereas 32 clusters differed between them that included a deletion of 18 508 bp containing mfs1, mao1, dmaT, and pks-nrps for the cyclopiazonic acid (CPA) biosynthesis, explaining the no productivity of CPA in A. sojae. The A. sojae NBRC4239 genome data will be useful to characterize functional features of the koji moulds used in Japanese industries.

Sato, Atsushi; Oshima, Kenshiro; Noguchi, Hideki; Ogawa, Masahiro; Takahashi, Tadashi; Oguma, Tetsuya; Koyama, Yasuji; Itoh, Takehiko; Hattori, Masahira; Hanya, Yoshiki



Occurrence of Soybean Sleeping Blight Caused by Septogloeum sojae in Korea  

PubMed Central

Sleeping blight was observed on soybean plants grown in Yanggu, Suwon and Geumsan from 2005 to 2011. Symptoms developed on stems and pods of affected soybean plants. Five fungal isolates were obtained from the diseased plants and identified as Septogloeum sojae based on their morphological, cultural and molecular characteristics. Pathogenicity of the fungus was confirmed on soybean plants by artificial inoculation. This is the first report of S. sojae causing sleeping blight in soybean plants in Korea.

Choi, Hyo Won; Lee, Young Kee; Lee, Sang Yeob; Shim, Hong Sik



Mitochondrial genome sequences and comparative genomics of Phytophthora ramorum and P. sojae  

Microsoft Academic Search

The sequences of the mitochondrial genomes of the oomycetes Phytophthora ramorum and P. sojae were determined during the course of complete nuclear genome sequencing (Tyler et al., Science, 313:1261,2006). Both mitochondrial\\u000a genomes are circular mapping, with sizes of 39,314 bp for P. ramorum and 42,977 bp for P. sojae. Each contains a total of 37 recognizable protein-encoding genes, 26 or 25 tRNAs (P.

Frank N. Martin; Douda Bensasson; Brett M. Tyler; Jeffrey L. Boore



Understanding nonaflatoxigenicity of Aspergillus sojae : a windfall of aflatoxin biosynthesis research  

Microsoft Academic Search

Aspergillus section Flavi includes aflatoxin-producing and nonproducing fungi. Aspergillus sojae is unable to produce aflatoxins and is generally recognized as safe for food fermentation. However, because of its taxonomical\\u000a relatedness to aflatoxin-producing Aspergillus parasiticus and A. flavus, it is necessary to decipher the underlying mechanisms for its inability to produce aflatoxins. This review addresses the\\u000a relationship between A. sojae and

Perng-Kuang Chang; Kenichiro Matsushima; Tadashi Takahashi; Jiujiang Yu; Keietsu Abe; Deepak Bhatnagar; Gwo-Fang Yuan; Yasuji Koyama; Thomas E. Cleveland



Heterologous expression of a pleiotropic drug resistance transporter from Phytophthora sojae in yeast transporter mutants  

Microsoft Academic Search

A system for the expression of an ATP binding cassette (ABC) transporter from the soybean pathogen Phytophthora sojae is described. Pdr1, an ABC transporter with homology to the pleiotropic drug resistance (PDR) family of transporters, was cloned by primer walking\\u000a from a P. sojae genomic library. Reverse transcriptase PCR assays showed that the transcript disappeared after encystment of zoospores and

Mary S. Connolly; Yasuko Sakihama; Vipaporn Phuntumart; Yinjun Jiang; Franklin Warren; Lindsay Mourant; Paul F. Morris



Molecular mapping and characterization of two genes conferring resistance to Phytophthora sojae in a soybean landrace PI 567139B  

Technology Transfer Automated Retrieval System (TEKTRAN)

Phytophthora root and stem rot (PRR), caused by the soil-borne oomycete pathogen Phytophthora sojae, is one of the most destructive diseases of soybean. PRR can be effectively controlled by race-specific genes conferring resistance to P. sojae (Rps). However, the Rps genes are usually non-durable, a...


Chemotactic Preferences and Strain Variation in the Response of Phytophthora sojae Zoospores to Host Isoflavones  

PubMed Central

The zoospores of Phytophthora sojae are chemotactically attracted to the isoflavones genistein and daidzein that are released by soybean roots. In this study we have examined the response of P. sojae zoospores to a wide range of compounds having some structural similarity to genistein and daidzein, including isoflavones, flavones, chalcones, stilbenes, benzoins, benzoates, benzophenones, acetophenones, and coumarins. Of 59 compounds examined, 43 elicited some response. A comparison of the chemotactic responses elicited by the various compounds revealed a primary role for the phenolic 4(prm1)- and 7-hydroxyl groups on the isoflavone structure. A few compounds acted as repellents, notably methylated flavones with a hydrophobic B ring. The chemotactic response to many of the analogs was markedly different among different strains of P. sojae.

Tyler, B. M.; Wu, M.; Wang, J.; Cheung, W.; Morris, P. F.



Genotypic Properties of the Entomopathogenic Fungus Nomuraea rileyi  

Microsoft Academic Search

The dimorphic entomopathogenic fungus Nomuraea rileyi, an important biological control agent of various noctuid defoliators, is known to regulate Anticarsia gemmatalis, the key soybean pest in South America and the southeastern United States. At present, very little is known about the isolates that are associated with the endemic or epizootic phases of this disease agent. The development of molecular markers

Drion G Boucias; Myrian S Tigano; Daniel R Sosa-Gomez; Travis R Glare; Peter W Inglis




Technology Transfer Automated Retrieval System (TEKTRAN)

Artificial diet was supplemented with avidin at 10 and 100 ppm to determine its effects on growth and mortality of five lepidopteran insects: Helicoverpa zea, Heliothis virescens, Spodoptera frugiperda, Spodoptera exigua, and Anticarsia gemmatalis. All insects were placed on diet immediately after...



Technology Transfer Automated Retrieval System (TEKTRAN)

Diflubenzuron [Dimilin, N-[{(4-chlorophenyl)amino}carbonyl]-2-6-difluorobenzamide] is an insect growth regulator that is highly effective and provides residual control (up to 54 days) against velvetbean caterpillar (Anticarsia gemmatalis Hübner) on soybean [Glycine max (L.) Merrill] in Louisiana. W...


Insect pests and yield potential of vegetable soybean (Endamame) produced in Georgia  

Technology Transfer Automated Retrieval System (TEKTRAN)

A series of replicated field experiments was conducted with vegetable soybean (edamame), Glycine max (L.) Merrill, to assess the impacts of cultivars, planting dates, and insecticidal controls on insect pest abundance, crop damage and yield potential. The velvetbean caterpillar, Anticarsia gemmatali...


Immunity of an Alternative Host Can Be Overcome by Higher Densities of Its Parasitoids Palmistichus elaeisis and Trichospilus diatraeae  

Microsoft Academic Search

Interactions of the parasitoids Palmistichus elaeisis Delvare & LaSalle and Trichospilus diatraeae Cherian & Margabandhu (Hymenoptera: Eulophidae) with its alternative host Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae) affect the success or failure of the mass production of these parasitoids for use in integrated pest management programs. The aim of this study was to evaluate changes in the cellular defense and encapsulation

Gilberto Santos Andrade; José Eduardo Serrão; José Cola Zanuncio; Teresinha Vinha Zanuncio; Germano Leão Demolin Leite; Ricardo Antonio Polanczyk



Understanding nonaflatoxigenicity of Aspergillus sojae: a windfall of aflatoxin biosynthesis research.  


Aspergillus section Flavi includes aflatoxin-producing and nonproducing fungi. Aspergillus sojae is unable to produce aflatoxins and is generally recognized as safe for food fermentation. However, because of its taxonomical relatedness to aflatoxin-producing Aspergillus parasiticus and A. flavus, it is necessary to decipher the underlying mechanisms for its inability to produce aflatoxins. This review addresses the relationship between A. sojae and A. parasiticus and the advances that have been made in aflatoxin biosynthesis research, especially with regard to gene structure, genome organization, and gene regulation in A. parasiticus and A. flavus and how this has been used to assure the safety of A. sojae as an organism for food fermentation. The lack of aflatoxin-producing ability of A. sojae results primarily from an early termination point mutation in the pathway-specific aflR regulatory gene, which causes the truncation of the transcriptional activation domain of AflR and the abolishment of interaction between AflR and the AflJ co-activator. Both are required for gene expression. In addition, a defect in the polyketide synthase gene also contributes to its nonaflatoxigenicity. PMID:17665189

Chang, Perng-Kuang; Matsushima, Kenichiro; Takahashi, Tadashi; Yu, Jiujiang; Abe, Keietsu; Bhatnagar, Deepak; Yuan, Gwo-Fang; Koyama, Yasuji; Cleveland, Thomas E



Fine-scale phylogenetic structure and major events in the history of the current wild soybean (Glycine soja) and taxonomic assignment of semi-wild type (Glycine gracilis Skvortz.) within the Chinese subgenus Soja.  


Wild and cultivated species of soybeans have coexisted for 5000 years in China. Despite this long history, there is very little information on the genetic relationship of Glycine soja and G. max. To gain insight into the major events in the history of the subgenus Soja, we examined 20 simple sequence repeat (SSR) markers of a large number of accessions (910). The results showed no significant differences between wild and semi-wild soybeans in genetic diversity but significant differences between G. soja and G. max. Ancestry and cluster analyses revealed that semi-wild soybeans should belong to the wild category and not to G. max. Our results also showed that differentiation had occurred not only among G. soja, G. gracilis, and G. max but also within G. soja and within G. gracilis. Glycine soja had 3 clear genetic categories: typical small-seeded (?2.0 g 100-seed weight), dual-origin middle-seeded (2.0-2.5 g), and large-seeded plants (2.51-3.0 g). These last were genetically close to G. gracilis, their defining some traits having been acquired mainly by introgression from soybeans. Small-seeded G. gracilis (3.01-3.5 g) were genetically different from larger seeded ones (from 3.51 to 4.0 to over 10 g). Seed size predominated over seed coat color in evolutionary degree. Typical and large-seeded G. soja were found to have 0.7% and 12% introgressive cultivar genes, respectively. The genetic boundary of G. gracilis was at the range of 2.51-3.0 g of G. soja. In the great majority of wild accessions, traits such as white flowers, gray pubescences, no-seed bloom, and colored seed coats were likely introgressive from domesticated soybeans. PMID:21984661

Wang, Ke-Jing; Li, Xiang-Hua; Liu, Yang



Evaluation of Soybean Cultivars with the Rps1k Gene for Partial Resistance or Field Tolerance to Phytophthora sojae  

Microsoft Academic Search

Phytophthora root rot, caused by Phytophthora sojae Kaufmann andGerdeman,primarilyattackstherootsofsoybean(Glycinemax(L.) Merr.) plants. Partial resistance and field tolerance in 14 commercial glyphosate (N-(phosphonomethyl)glycine) tolerant soybean cultivars with the Rps1k resistance gene were studied. Partial resistance to compatible P. sojae races 28 and 30 was evaluated by the agar layer technique. Relative to the percentage of the control, all of the com- mercial

C. R. Ferro; C. B. Hill; M. R. Miles; G. L. Hartman


Copy Number Variation and Transcriptional Polymorphisms of Phytophthora sojae RXLR Effector Genes Avr1a and Avr3a  

Microsoft Academic Search

The importance of segmental duplications and copy number variants as a source of genetic and phenotypic variation is gaining greater appreciation, in a variety of organisms. Now, we have identified the Phytophthora sojae avirulence genes Avr1a and Avr3a and demonstrate how each of these Avr genes display copy number variation in different strains of P. sojae. The Avr1a locus is

Dinah Qutob; Jennifer Tedman-Jones; Suomeng Dong; Kuflom Kuflu; Hai Pham; Yuanchao Wang; Daolong Dou; Shiv D. Kale; Felipe D. Arredondo; Brett M. Tyler; Mark Gijzen; Frederick M. Ausubel



Non-toxic albumin and soja protein borates as ground-contact wood preservatives  

Microsoft Academic Search

Wood preservatives based on protein borates, both obtained by just mixed water solutions of protein and boric acid, as well\\u000a as in the case of premanufactured protein borates salts are shown to be a good method to greatly retard the leaching of boron\\u000a from treated timber. Hence just premixed albumin + boric acid, premanufactured albumin borate and soja protein +

M.-F. Thevenon; A. Pizzi; J.-P. Haluk



Global transcriptome profiling of wild soybean (Glycine soja) roots under NaHCO3 treatment  

Microsoft Academic Search

BACKGROUND: Plant roots are the primary site of perception and injury for saline-alkaline stress. The current knowledge of saline-alkaline stress transcriptome is mostly focused on saline (NaCl) stress and only limited information on alkaline (NaHCO3) stress is available. RESULTS: Using Affymetrix® Soybean GeneChip®, we conducted transcriptional profiling on Glycine soja roots subjected to 50 mmol\\/L NaHCO3 treatment. In a total

Ying Ge; Yong Li; Yan-Ming Zhu; Xi Bai; De-Kang Lv; Dianjing Guo; Wei Ji; Hua Cai



Dissection of two soybean QTL conferring partial resistance to Phytophthora sojae through sequence and gene expression analysis  

PubMed Central

Background Phytophthora sojae is the primary pathogen of soybeans that are grown on poorly drained soils. Race-specific resistance to P. sojae in soybean is gene-for-gene, although in many areas of the US and worldwide there are populations that have adapted to the most commonly deployed resistance to P. sojae ( Rps) genes. Hence, this system has received increased attention towards identifying mechanisms and molecular markers associated with partial resistance to this pathogen. Several quantitative trait loci (QTL) have been identified in the soybean cultivar ‘Conrad’ that contributes to the expression of partial resistance to multiple P. sojae isolates. Results In this study, two of the Conrad QTL on chromosome 19 were dissected through sequence and expression analysis of genes in both resistant (Conrad) and susceptible (‘Sloan’) genotypes. There were 1025 single nucleotide polymorphisms (SNPs) in 87 of 153 genes sequenced from Conrad and Sloan. There were 304 SNPs in 54 genes sequenced from Conrad compared to those from both Sloan and Williams 82, of which 11 genes had SNPs unique to Conrad. Eleven of 19 genes in these regions analyzed with qRT-PCR had significant differences in fold change of transcript abundance in response to infection with P. sojae in lines with QTL haplotype from the resistant parent compared to those with the susceptible parent haplotype. From these, 8 of the 11 genes had SNPs in the upstream, untranslated region, exon, intron, and/or downstream region. These 11 candidate genes encode proteins potentially involved in signal transduction, hormone-mediated pathways, plant cell structural modification, ubiquitination, and basal resistance. Conclusions These findings may indicate a complex defense network with multiple mechanisms underlying these two soybean QTL conferring resistance to P. sojae. SNP markers derived from these candidate genes can contribute to fine mapping of QTL and marker assisted breeding for resistance to P. sojae.



Cross-species global proteomics reveals conserved and unique processes in Phytophthora sojae and P. ramorum  

SciTech Connect

Phytophthora ramorum and Phytophthora sojae are destructive plant pathogens. Phytophthora sojae has a narrow host range whereas P. ramorum has a wide host range. A global proteomic comparison of the vegetative (mycelium) and infective (germinating-cyst) life-stages of P. sojae and P. ramorum was conducted to identify candidate proteins involved in host range, early infection and vegetative growth. Sixty-two candidates for early infection, 26 candidates for vegetative growth, and numerous proteins that may be involved in defining host specificity were identified. In addition, common life stage proteomic trends between the organisms were observed. In mycelia, proteins involved in transport and metabolism of amino acids, carbohydrates and other small molecules were up-regulated. In the germinating cysts, up-regulated proteins associated with lipid transport and metabolism, cytoskeleton and protein synthesis were observed. It appears that the germinating cyst catabolizes lipid reserves through the -oxidation pathway to drive the extensive protein synthesis necessary to produce the germ tube and initiate infection. Once inside the host, the pathogen switches to vegetative growth, where energy is derived from glycolysis and utilized for synthesis of amino acids and other molecules that assist survival in the plant tissue.

Savidor, Alon [ORNL; Donahoo, Ryan S [ORNL; Hurtado-Gonzales, Oscar [University of Tennessee, Knoxville (UTK); Land, Miriam L [ORNL; Shah, Manesh B [ORNL; Lamour, Kurt H [ORNL; McDonald, W Hayes [ORNL



Genetic and physical mapping of Avr1a in Phytophthora sojae.  

PubMed Central

The interaction between soybean and the phytopathogenic oomycete Phytophthora sojae is controlled by host resistance (Rps) genes and pathogen avirulence (Avr) genes. We have mapped the Avr1a locus in F(2) populations derived from four different P. sojae races. Four RAPD and nine AFLP markers linked to Avr1a were initially identified. Nine markers were used to compare genetic linkage maps of the Avr1a locus in two distinct F(2) populations. Distorted segregation ratios favoring homozygous genotypes were noted in both crosses. Segregation analysis of all the markers in one F(2) population of 90 progeny generated a map of 113.2 cM encompassing Avr1a, with one marker cosegregating with the gene. The cosegregating DNA marker was used to isolate P. sojae BAC clones and construct a physical map covering 170 kb, from which additional DNA markers were developed. Three markers occurring within the BAC contig were mapped in an enlarged population of 486 F(2) progeny. Avr1a was localized to a 114-kb interval, and an average physical to genetic distance ratio of 391 kb/cM was calculated for this region. This work provides a basis for the positional cloning of Avr1a.

MacGregor, Terry; Bhattacharyya, Madan; Tyler, Brett; Bhat, Ravindra; Schmitthenner, August F; Gijzen, Mark



Genetic analysis and identification of DNA markers linked to a novel Phytophthora sojae resistance gene in the Japanese soybean cultivar Waseshiroge  

Technology Transfer Automated Retrieval System (TEKTRAN)

Glycine max (L.) Merr. cv. Waseshiroge is considered to be strongly resistant to several races of Phytophthora sojae in Japan. In order to characterize the inheritance of Waseshiroge resistance to P. sojae isolates, 42 F2 progeny plants and 94 F7:8 families were produced from crosses between the sus...


Two RxLR avirulence genes in Phytophthora sojae determine soybean Rps1k-mediated disease resistance.  


Resistance to Phytophthora sojae (Rps) genes have been widely used in soybean against root and stem rot diseases caused by this oomycete. Among 15 known soybean Rps genes, Rps1k has been the most widely used in the past four decades. Here, we show that the products of two distinct but closely linked RxLR effector genes are detected by Rps1k-containing plants, resulting in disease resistance. One of the genes is Avr1b-1, that confers avirulence in the presence of Rps1b. Three lines of evidence, including overexpression and gene silencing of Avr1b-1 in stable P. sojae transformants, as well as transient expression of this gene in soybean, indicated that Avr1b could trigger an Rps1k-mediated defense response. Some isolates of P. sojae that do not express Avr1b are nevertheless unable to infect Rps1k plants. In those isolates, we identified a second RxLR effector gene (designated Avr1k), located 5 kb away from Avr1b-1. Silencing or overexpression of Avr1k in P. sojae stable transformants resulted in the loss or gain, respectively, of the avirulence phenotype in the presence of Rps1k. Only isolates of P. sojae with mutant alleles of both Avr1b-1 and Avr1k could evade perception by the soybean plants carrying Rps1k. PMID:23530601

Song, Tianqiao; Kale, Shiv D; Arredondo, Felipe D; Shen, Danyu; Su, Liming; Liu, Li; Wu, Yuren; Wang, Yuanchao; Dou, Daolong; Tyler, Brett M



Enhanced gene targeting frequency in ku70 and ku80 disruption mutants of Aspergillus sojae and Aspergillus oryzae.  


In the koji molds Aspergillus sojae and Aspergillus oryzae, exogenous DNA is integrated in the genome, in most cases irrespective of the sequence homology, suggesting that DNA integration occurs predominantly through a nonhomologous end joining pathway where two ku genes, namely, ku70 and ku80, play a key role. To determine the effect of ku gene disruption on the gene targeting frequency, we constructed ku70-, ku80-, and ku70-ku80-disrupted strains of A. sojae and A. oryzae. The gene targeting frequency of the tannase gene in ku70 and ku80 strains of both Aspergillus species was markedly enhanced as compared with that of the parental strains. The gene targeting frequency of the aflR and ku80 genes was also enhanced in an A. sojae ku70 background. Therefore, the koji mold strains with ku-disrupted genes will be excellent tools as hosts for efficient gene targeting. PMID:16470383

Takahashi, Tadashi; Masuda, Tsutomu; Koyama, Yasuji



Isozyme differentiation among three pathotypes of the entomogenous fungus Nomuraea rileyi.  


The zymogram technique has been applied to three pathotypes of the entomopathogenic fungus Nomuraea rileyi. Isozyme profiles of isolates from Heliothis zea, Pseudoplusia includens, and Anticarsia gemmatalis were compared for 17 enzymes of known metabolic function. Electrophoretic data supported the taxonomic differences inferred for the three pathotypes based on host specificity. The isolate from A. gemmatalis was found consistently to be the most distinct. This study demonstrates that isozyme analysis may be used to distinguish closely related fungal isolates of N. rileyi. PMID:7195298

Joslyn, D J; Boucias, D G



Identification of ?-tocopherol and ?-tocopheryl acetate from the cuticle of soybean pods armyworm (Spodoptera cosmioides).  


The chemical composition of the soybean pods armyworm Spodoptera cosmioides (Walker, 1858) (Lepidoptera: Noctuidae) and Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Erebidae) larval cuticles was evaluated using gas chromatography coupled to a mass detector (GC-MS). Among the usual lipids found in the insect cuticle, ?-tocopherol and ?-tocopheryl acetate were also isolated from S. cosmioides. On the other hand, no vitamin E derivative was found in A. gemmatalis exuvia. This is the first report of vitamin E occurrence in the insect's cuticle. PMID:23356865

Fronza, Edegar; Migues, Ignacio; Specht, Alexandre; de Barros, Neiva Monteiro; Heinzen, Horacio



Diamagnetic Susceptibility and Root Growth Responses to Magnetic Fields in Lens culinaris, Glycine soja , and Triticum aestivum  

Microsoft Academic Search

We studied the response of root growth to different magnetic fields and forces. We submitted the seeds of three plant species, Lens culinaris L., Glycine soja Siebold & Zucc., and Triticum aestivum L., which differ in concentrations of paramagnetic (e.g., Fe or Co) and diamagnetic materials (e.g., starchy amyloplasts), to different static magnetic fields and forces. A magnetic field of

Josep Peñuelas; Joan Llusià; Benjamín Martínez; Josep Fontcuberta



The NLP toxin family in Phytophthora sojae includes rapidly evolving groups that lack necrosis-inducing activity.  


Necrosis- and ethylene-inducing-like proteins (NLP) are widely distributed in eukaryotic and prokaryotic plant pathogens and are considered to be important virulence factors. We identified, in total, 70 potential Phytophthora sojae NLP genes but 37 were designated as pseudogenes. Sequence alignment of the remaining 33 NLP delineated six groups. Three of these groups include proteins with an intact heptapeptide (Gly-His-Arg-His-Asp-Trp-Glu) motif, which is important for necrosis-inducing activity, whereas the motif is not conserved in the other groups. In total, 19 representative NLP genes were assessed for necrosis-inducing activity by heterologous expression in Nicotiana benthamiana. Surprisingly, only eight genes triggered cell death. The expression of the NLP genes in P. sojae was examined, distinguishing 20 expressed and 13 nonexpressed NLP genes. Real-time reverse-transcriptase polymerase chain reaction results indicate that most NLP are highly expressed during cyst germination and infection stages. Amino acid substitution ratios (Ka/Ks) of 33 NLP sequences from four different P. sojae strains resulted in identification of positive selection sites in a distinct NLP group. Overall, our study indicates that expansion and pseudogenization of the P. sojae NLP family results from an ongoing birth-and-death process, and that varying patterns of expression, necrosis-inducing activity, and positive selection suggest that NLP have diversified in function. PMID:22397404

Dong, Suomeng; Kong, Guanghui; Qutob, Dinah; Yu, Xiaoli; Tang, Junli; Kang, Jixiong; Dai, Tingting; Wang, Hai; Gijzen, Mark; Wang, Yuanchao



Identification of the Glutaminase Genes of Aspergillus sojae Involved in Glutamate Production during Soy Sauce Fermentation.  


Glutaminase, an enzyme that catalyzes the conversion of L-glutamine to L-glutamate, enhances the umami taste in soy sauce. The Aspergillus sojae genome contains 10 glutaminase genes. In this study, we estimated that approximately 60% of the glutamate in soy sauce is produced through the glutaminase reaction. To determine which glutaminase is involved in soy sauce glutamate production, we prepared soy sauces using single and multiple glutaminase gene disruptants of A. sojae. The glutamate concentration in soy sauce prepared using the ?gahA-?gahB-?ggtA-?gls disruptant was approximately 60% lower than that in the control strain, whereas it was decreased by approximately 20-30% in the ?gahA-?gahB disruptant. However, the glutamate concentration was unchanged in the soy sauces prepared using the ?gahA-?ggtA-?gls and ?gahB-?ggtA-?gls disruptants. These results indicate that four glutaminases are involved in glutamate production in soy sauce, and that the peptidoglutaminase activities of GahA and GahB increase the glutamate concentration in soy sauce. PMID:24018673

Ito, Kotaro; Koyama, Yasuji; Hanya, Yoshiki



Molecular mapping of two genes conferring resistance to Phytophthora sojae in a soybean landrace PI 567139B.  


Phytophthora root and stem rot (PRR), caused by the soil-borne oomycete pathogen Phytophthora sojae, is one of the most destructive diseases of soybean. PRR can be effectively controlled by race-specific genes conferring resistance to P. sojae (Rps). However, the Rps genes are usually non-durable, as populations of P. sojae are highly diverse and quick to adapt, and can be overcome 8-15 years after deployment. Thus, it is important to identify novel Rps genes for development of resistant soybean cultivars. PI 567139B is a soybean landrace carrying excellent resistance to nearly all predominant P. sojae races in Indiana. A mapping population consisting of 245 F2 individuals and 403 F2:3 families was developed from a cross between PI 567139B and the susceptible cultivar 'Williams', and used to dissect the resistance carried by PI 567139B. We found that the resistance in PI 567139B was conferred by two independent Rps genes, designated RpsUN1 and RpsUN2. The former was mapped to a 6.5 cM region between SSR markers Satt159 and BARCSOYSSR_03_0250 that spans the Rps1 locus on chromosome 3, while the latter was mapped to a 3.0 cM region between BARCSOYSSR_16_1275 and Sat_144, approximately 3.0-3.4 cM upstream of Rps2 on chromosome 16. According to the 'Williams 82' reference genome sequence, both regions are highly enriched with NBS-LRR genes. Marker assisted resistance spectrum analyses of these genes with 16 isolates of P. sojae, in combination with the mapping results, suggested that RpsUN1 was likely to be a novel allele at the Rps1 locus, while RpsUN2 was more likely to be a novel Rps gene. PMID:23689748

Lin, Feng; Zhao, Meixia; Ping, Jieqing; Johnson, Austin; Zhang, Biao; Abney, T Scott; Hughes, Teresa J; Ma, Jianxin



Cell Culture Derived AgMNPV Bioinsecticide: Biological Constraints and Bioprocess Issues  

Microsoft Academic Search

We have studied parameters for optimizing the Spodoptera frugiperda (Sf9) cell culture and viral infection for the production of Anticarsia gemmatalis multiple nucleopolyhedrosis virus (AgMNPV) polyhedra inclusion bodies (PIBs) in shaker-Schott or spinner bottles and bioreactors.\\u000a We have assayed the kLa of the systems, initial cell seeding, cell culture volume, dissolved oxygen (DO), multiplicity of infection (MOI), nutrients\\u000a consumption, and

Valeria M. Rodas; Fabiano H. Marques; Marcelo T. Honda; Daniela M. Soares; Soraia A. C. Jorge; Marta M. Antoniazzi; Claudia Medugno; Maria E. B. Castro; Bergmann M. Ribeiro; Marlinda L. Souza; Aldo Tonso; Carlos A. Pereira



Glyceollin is an Important Component of Soybean Plant Defense Against Phytophthora sojae and Macrophomina phaseolina.  


ABSTRACT The response of soybean transgenic plants, with suppressed synthesis of isoflavones, and nontransgenic plants to two common soybean pathogens, Macrophomina phaseolina and Phytophthora sojae, was studied. Transgenic soybean plants of one line used in this study were previously generated via bombardment of embryogenic cultures with the phenylalanine ammonia lyase, chalcone synthase, and isoflavone synthase (IFS2) genes in sense orientation driven by the cotyledon-preferable lectin promoter (to turn genes on in cotyledons), while plants of another line were newly produced using the IFS2 gene in sense orientation driven by the Cassava vein mosaic virus constitutive promoter (to turn genes on in all plant parts). Nearly complete inhibition of isoflavone synthesis was found in the cotyledons of young seedlings of transgenic plants transformed with the IFS2 transgene driven by the cotyledon-preferable lectin promoter compared with the untransformed control during the 10-day observation period, with the precursors of isoflavone synthesis being accumulated in the cotyledons of transgenic plants. These results indicated that the lectin promoter could be active not only during seed development but also during seed germination. Downregulation of isoflavone synthesis only in the seed or in the whole soybean plant caused a strong inhibition of the pathogen-inducible glyceollin in cotyledons after inoculation with P. sojae, which resulted in increased susceptibility of the cotyledons of both transgenic lines to this pathogen compared with inoculated cotyledons of untransformed plants. When stems were inoculated with M. phaseolina, suppression of glyceollin synthesis was found only in stems of transgenic plants expressing the transgene driven by a constitutive promoter, which developed more severe infection. These results provide further evidence that rapid glyceollin accumulation during infection contributes to the innate soybean defense response. PMID:23617338

Lygin, Anatoliy V; Zernova, Olga V; Hill, Curtis B; Kholina, Nadegda A; Widholm, Jack M; Hartman, Glen L; Lozovaya, Vera V



Identification of wild soybean (Glycine soja) TIFY family genes and their expression profiling analysis under bicarbonate stress.  


Wild soybean (Glycine soja L. G07256) exhibits a greater adaptability to soil bicarbonate stress than cultivated soybean, and recent discoveries show that TIFY family genes are involved in the response to several abiotic stresses. A genomic and transcriptomic analysis of all TIFY genes in G. soja, compared with G. max, will provide insight into the function of this gene family in plant bicarbonate stress response. This article identified and characterized 34 TIFY genes in G. soja. Sequence analyses indicated that most GsTIFY proteins had two conserved domains: TIFY and Jas. Phylogenetic analyses suggested that these GsTIFY genes could be classified into two groups. A clustering analysis of all GsTIFY transcript expression profiles from bicarbonate stress treated G. soja showed that there were five different transcript patterns in leaves and six different transcript patterns in roots when the GsTIFY family responds to bicarbonate stress. Moreover, the expression level changes of all TIFY genes in cultivated soybean, treated with bicarbonate stress, were also verified. The expression comparison analysis of TIFYs between wild and cultivated soybeans confirmed that, different from the cultivated soybean, GsTIFY (10a, 10b, 10c, 10d, 10e, 10f, 11a, and 11b) were dramatically up-regulated at the early stage of stress, while GsTIFY 1c and 2b were significantly up-regulated at the later period of stress. The frequently stress responsive and diverse expression profiles of the GsTIFY gene family suggests that this family may play important roles in plant environmental stress responses and adaptation. PMID:23090726

Zhu, Dan; Bai, Xi; Luo, Xiao; Chen, Qin; Cai, Hua; Ji, Wei; Zhu, Yanming



Alkaline-stress response in Glycine soja leaf identifies specific transcription factors and ABA-mediated signaling factors  

Microsoft Academic Search

Transcriptome of Glycine soja leaf tissue during a detailed time course formed a foundation for examining transcriptional processes during NaHCO3 stress treatment. Of a total of 2,310 detected differentially expressed genes, 1,664 genes were upregulated and 1,704 genes\\u000a were downregulated at various time points. The number of stress-regulated genes increased dramatically after a 6-h stress\\u000a treatment. GO category gene enrichment

Ying Ge; Yong Li; De-Kang Lv; Xi Bai; Wei Ji; Hua Cai; Ao-Xue Wang; Yan-Ming Zhu



The Phytophthora sojae Avirulence Locus Avr3c Encodes a Multi-Copy RXLR Effector with Sequence Polymorphisms among Pathogen Strains  

PubMed Central

Root and stem rot disease of soybean is caused by the oomycete Phytophthora sojae. The avirulence (Avr) genes of P. sojae control race-cultivar compatibility. In this study, we identify the P. sojae Avr3c gene and show that it encodes a predicted RXLR effector protein of 220 amino acids. Sequence and transcriptional data were compared for predicted RXLR effectors occurring in the vicinity of Avr4/6, as genetic linkage of Avr3c and Avr4/6 was previously suggested. Mapping of DNA markers in a F2 population was performed to determine whether selected RXLR effector genes co-segregate with the Avr3c phenotype. The results pointed to one RXLR candidate gene as likely to encode Avr3c. This was verified by testing selected genes by a co-bombardment assay on soybean plants with Rps3c, thus demonstrating functionality and confirming the identity of Avr3c. The Avr3c gene together with eight other predicted genes are part of a repetitive segment of 33.7 kb. Three near-identical copies of this segment occur in a tandem array. In P. sojae strain P6497, two identical copies of Avr3c occur within the repeated segments whereas the third copy of this RXLR effector has diverged in sequence. The Avr3c gene is expressed during the early stages of infection in all P. sojae strains examined. Virulent alleles of Avr3c that differ in amino acid sequence were identified in other strains of P. sojae. Gain of virulence was acquired through mutation and subsequent sequence exchanges between the two copies of Avr3c. The results illustrate the importance of segmental duplications and RXLR effector evolution in the control of race-cultivar compatibility in the P. sojae and soybean interaction.

Dong, Suomeng; Qutob, Dinah; Tedman-Jones, Jennifer; Kuflu, Kuflom; Wang, Yuanchao; Tyler, Brett M.; Gijzen, Mark



A Myb transcription factor of Phytophthora sojae, regulated by MAP kinase PsSAK1, is required for zoospore development.  


PsSAK1, a mitogen-activated protein (MAP) kinase from Phytophthora sojae, plays an important role in host infection and zoospore viability. However, the downstream mechanism of PsSAK1 remains unclear. In this study, the 3'-tag digital gene expression (DGE) profiling method was applied to sequence the global transcriptional sequence of PsSAK1-silenced mutants during the cysts stage and 1.5 h after inoculation onto susceptible soybean leaf tissues. Compared with the gene expression levels of the recipient P. sojae strain, several candidates of Myb family were differentially expressed (up or down) in response to the loss of PsSAK1, including of a R2R3-type Myb transcription factor, PsMYB1. qRT-PCR indicated that the transcriptional level of PsMYB1 decreased due to PsSAK1 silencing. The transcriptional level of PsMYB1 increased during sporulating hyphae, in germinated cysts, and early infection. Silencing of PsMYB1 results in three phenotypes: a) no cleavage of the cytoplasm into uninucleate zoospores or release of normal zoospores, b) direct germination of sporangia, and c) afunction in zoospore-mediated plant infection. Our data indicate that the PsMYB1 transcription factor functions downstream of MAP kinase PsSAK1 and is required for zoospore development of P. sojae. PMID:22768262

Zhang, Meng; Lu, Jing; Tao, Kai; Ye, Wenwu; Li, Aining; Liu, Xiaoyun; Kong, Liang; Dong, Suomeng; Zheng, Xiaobo; Wang, Yuanchao



Cloning, expression and characterization of endo-beta-1,4-mannanase from Aspergillus fumigatus in Aspergillus sojae and Pichia pastoris.  


To be utilized in biomass conversion, including ethanol production and galactosylated oligosaccharide synthesis, namely prebiotics, the gene of extracellular endo-beta-1,4-mannanase (EC of Aspergillus fumigatus IMI 385708 (formerly known as Thermomyces lanuginosus IMI 158749) was expressed first in Aspergillus sojae and then in Pichia pastoris under the control of the glyceraldehyde triphosphate dehydrogenase (gpdA) and the alcohol oxidase (AOX1) promoters, respectively. The highest production of mannanase (352 U mL(-1)) in A. sojae was observed after 6 days of cultivation. In P. pastoris, the highest mannanase production was observed 10 h after induction with methanol (61 U mL(-1)). The fold increase in mannanase production was estimated as approximately 12-fold and approximately 2-fold in A. sojae and P. pastoris, respectively, when compared with A. fumigatus. Both recombinant enzymes showed molecular mass of about 60 kDa and similar specific activities ( approximately 350 U mg(-1) protein). Temperature optima were at 60 degrees C and 45 degrees C, and maximum activity was at pH 4.5 and 5.2 for A. sojae and P. pastoris, respectively. The enzyme from P. pastoris was more stable retaining most of the activity up to 50 degrees C, whereas the enzyme from A. sojae rapidly lost activity above 40 degrees C. PMID:19205049

Duruksu, Gökhan; Ozturk, Bengu; Biely, Peter; Bakir, Ufuk; Ogel, Zumrut B


Global transcriptome profiling of wild soybean (Glycine soja) roots under NaHCO3 treatment  

PubMed Central

Background Plant roots are the primary site of perception and injury for saline-alkaline stress. The current knowledge of saline-alkaline stress transcriptome is mostly focused on saline (NaCl) stress and only limited information on alkaline (NaHCO3) stress is available. Results Using Affymetrix® Soybean GeneChip®, we conducted transcriptional profiling on Glycine soja roots subjected to 50 mmol/L NaHCO3 treatment. In a total of 7088 probe sets, 3307 were up-regulated and 5720 were down-regulated at various time points. The number of significantly stress regulated genes increased dramatically after 3 h stress treatment and peaked at 6 h. GO enrichment test revealed that most of the differentially expressed genes were involved in signal transduction, energy, transcription, secondary metabolism, transporter, disease and defence response. We also detected 11 microRNAs regulated by NaHCO3 stress. Conclusions This is the first comprehensive wild soybean root transcriptome analysis under alkaline stress. These analyses have identified an inventory of genes with altered expression regulated by alkaline stress. The data extend the current understanding of wild soybean alkali stress response by providing a set of robustly selected, differentially expressed genes for further investigation.



Phylogenetic relationships of the mitochondrial genomes in the genus Glycine subgenus Soja.  


Restriction fragment length polymorphisms (RFLPs) of mitochondrial DNA (mtDNA) of wild and cultivated soybeans were analyzed to study their phylogenetic relationships. The observed number of differences in hybridization profiles greatly varied (1-17 patterns) with both the mtDNA probes and the restriction enzymes that were used. A cladistic analysis was conducted based on the RFLP data. In the parsimonious tree, four distinct groups appeared among 20 accessions of the subgenus Soja representing 20 mitochondrial genome types. Common features with regard to geographic distributions in natural populations in East Asia were observed among the mitochondrial genome types of wild soybean that belonged to the same group: one clade consisted of genome types IIg and VIIg that are detected with very low frequencies; another clade consisted of genome types Ic, Id, Ie, and Ik whose distributions are highly biased mainly in Japan. The genome types that are widely distributed in East Asia such as IVa, IVb, and Va were not grouped into the same clade. The mitochondrial genome types IIIb, IVb, and IVc, in which two different chloroplast genome types exist, belonged to the same clade. Possible changes in mitochondrial genomes during the expansion of the distribution of wild soybeans in East Asia were discussed. PMID:9880923

Kanazawa, A; Tozuka, A; Akimoto, S; Abe, J; Shimamoto, Y



The Effect of Potassium Nitrate on the Reduction of Phytophthora Stem Rot Disease of Soybeans, the Growth Rate and Zoospore Release of Phytophthora Sojae  

Technology Transfer Automated Retrieval System (TEKTRAN)

The effects of potassium nitrate (KNO3) application on Phytophthora stem rot disease reduction of Glycine max (L.) Merr. cvs. Chusei-Hikarikuro and Sachiyutaka, and fungal growth and zoospore release of a Phytophthora sojae isolate were investigated under laboratory conditions. The application of 4-...


The PsCZF1 gene encoding a C2H2 zinc finger protein is required for growth, development and pathogenesis in Phytophthora sojae.  


The C(2)H(2) zinc finger proteins form one of the largest families of transcriptional regulators in eukaryotes. We identified a Phytophthora sojae C(2)H(2) zinc finger (PsCZF1), that is highly conserved in sequenced oomycete pathogens. In transformants of P. sojae containing the PsCZF1 promoter fused to the beta-glucuronidase (GUS) reporter gene, GUS activity was highly induced in the P. sojae oospore stage and upregulated after infection. To elucidate the function of PsCZF1, its expression was silenced by introducing anti-sense constructs into P sojae. PsCZF1-silenced transformants did not exhibit altered cell size or morphology of sporangia and hyphae; however, hyphal growth rate was reduced by around 50% in the mutants. PsCZF1-deficient mutants were also impaired in production of oospores, swimming zoospores and germinating cysts, indicating that the gene is involved in various stages of the life cycle. Furthermore, we found that PsCZF1-deficient mutants lost virulence on host soybean cultivars. Our results suggest that this oomycete-specific C(2)H(2)-type zinc finger protein plays an important role in growth, development, and pathogenesis; therefore, PsCZF1 might be an attractive oomycete-specific target for chemical fungicide screening. PMID:19447167

Wang, Yonglin; Dou, Daolong; Wang, Xiaoli; Li, Aining; Sheng, Yuting; Hua, Chenlei; Cheng, Binyan; Chen, Xiaoren; Zheng, Xiaobo; Wang, Yuanchao



Identification of Candidate Signaling Genes Including Regulators of Chromosome Condensation 1 Protein Family Differentially Expressed in the Soybean - Phytophthora Sojae Interaction  

Technology Transfer Automated Retrieval System (TEKTRAN)

Stem and root rot caused by the oomycete pathogen, Phytopthora sojae, is a serious soybean disease. Use of Phytophthora resistance genes (Rps) in soybean cultivars has been very effective in controlling this pathogen. Resistance encoded by Rps genes is manifested through activation of defense resp...


GPR11, a putative seven-transmembrane G protein-coupled receptor, controls zoospore development and virulence of Phytophthora sojae.  


G protein-coupled receptors (GPCRs) represent a large receptor family involved in a broad spectrum of cell signaling. To understand signaling mechanisms mediated by GPCRs in Phytophthora sojae, we identified and characterized the PsGPR11 gene, which encodes a putative seven-transmembrane GPCR. An expression analysis revealed that PsGPR11 was differentially expressed during asexual development. The highest expression level occurred in zoospores and was upregulated during early infection. PsGPR11-deficienct transformants were obtained by gene silencing strategies. Silenced transformants exhibited no differences in hyphal growth or morphology, sporangium production or size, or mating behavior. However, the release of zoospores from sporangia was severely impaired in the silenced transformants, and about 50% of the sporangia did not completely release their zoospores. Zoospore encystment and germination were also impaired, and zoospores of the transformants lost their pathogenicity to soybean. In addition, no interaction was observed between PsGPR11 and PsGPA1 with a conventional yeast two-hybrid assay, and the transcriptional levels of some genes which were identified as being negatively regulated by PsGPA1 were not clearly altered in PsGPR11-silenced mutants. These results suggest that PsGPR11-mediated signaling controls P. sojae zoospore development and virulence through the pathways independent of G protein. PMID:20008081

Wang, Yonglin; Li, Aining; Wang, Xiaoli; Zhang, Xin; Zhao, Wei; Dou, Daolong; Zheng, Xiaobo; Wang, Yuanchao



PsVPS1, a dynamin-related protein, is involved in cyst germination and soybean infection of Phytophthora sojae.  


Plant pathogens secrete effector proteins to suppress plant immunity. However, the mechanism by which oomycete pathogens deliver effector proteins during plant infection remains unknown. In this report, we characterized a Phytophthora sojae vps1 gene. This gene encodes a homolog of the Saccharomyces cerevisiae vacuolar protein sorting gene vps1 that mediates budding of clathrin-coated vesicles from the late Golgi, which are diverted from the general secretory pathway to the vacuole. PsVPS1-silenced mutants were generated using polyethylene glycol-mediated protoplast stable transformation and were viable but had reduced extracellular protein activity. The PsVPS1-silenced mutants showed impaired hyphal growth, and the shapes of the vacuoles were highly fragmented. Silencing of PsVPS1 affected cyst germination as well as the polarized growth of germinated cysts. Silenced mutants showed impaired invasion of susceptible soybean plants regardless of wounding. These results suggest that PsVPS1 is involved in vacuole morphology and cyst development. Moreover, it is essential for the virulence of P. sojae and extracellular protein secretion. PMID:23516518

Li, Delong; Zhao, Zhijian; Huang, Yidan; Lu, Zhaojun; Yao, Meng; Hao, Yujuan; Zhai, Chunhua; Wang, Yuanchao



Genetic diversity and peculiarity of annual wild soybean (G. soja Sieb. et Zucc.) from various eco-regions in China.  


Annual wild soybean (Glycine soja Sieb. et Zucc.) is believed to be a potential gene source for future soybean improvement in coping with the world climate change for food security. To evaluate the wild soybean genetic diversity and differentiation, we analyzed allelic profiles at 60 simple-sequence repeat (SSR) loci and variation of eight morph-biological traits of a representative sample with 196 accessions from the natural growing area in China. For comparison, a representative sample with 200 landraces of Chinese cultivated soybean was included in this study. The SSR loci produced 1,067 alleles (17.8 per locus) with a mean gene diversity of 0.857 in the wild sample, which indicated the genetic diversity of G. soja was much higher than that of its cultivated counterpart (total 826 alleles, 13.7 per locus, mean gene diversity 0.727). After domestication, the genetic diversity of the cultigens decreased, with its 65.5% alleles inherited from the wild soybean, while 34.5% alleles newly emerged. AMOVA analysis showed that significant variance did exist among Northeast China, Huang-Huai-Hai Valleys and Southern China subpopulations. UPGMA cluster analysis indicated very significant association between the geographic grouping and genetic clustering, which demonstrated the geographic differentiation of the wild population had its relevant genetic bases. In comparison with the other two subpopulations, the Southern China subpopulation showed the highest allelic richness, diversity index and largest number of specific-present alleles, which suggests Southern China should be the major center of diversity for annual wild soybean. PMID:19449177

Wen, Zixiang; Ding, Yanlai; Zhao, Tuanjie; Gai, Junyi



Integration of the Draft Sequence and Physical Map as a Framework for Genomic Research in Soybean (Glycine max (L.) Merr.) and Wild Soybean (Glycine soja Sieb. and Zucc.)  

PubMed Central

Soybean is a model for the legume research community because of its importance as a crop, densely populated genetic maps, and the availability of a genome sequence. Even though a whole-genome shotgun sequence and bacterial artificial chromosome (BAC) libraries are available, a high-resolution, chromosome-based physical map linked to the sequence assemblies is still needed for whole-genome alignments and to facilitate map-based gene cloning. Three independent G. max BAC libraries combined with genetic and gene-based markers were used to construct a minimum tiling path (MTP) of BAC clones. A total of 107,214 clones were assembled into 1355 FPC (FingerPrinted Contigs) contigs, incorporating 4628 markers and aligned to the G. max reference genome sequence using BAC end-sequence information. Four different MTPs were made for G. max that covered from 92.6% to 95.0% of the soybean draft genome sequence (gmax1.01). Because our purpose was to pick the most reliable and complete MTP, and not the MTP with the minimal number of clones, the FPC map and draft sequence were integrated and clones with unpaired BES were added to build a high-quality physical map with the fewest gaps possible ( A physical map was also constructed for the undomesticated ancestor (G. soja) of soybean to explore genome variation between G. max and G. soja. 66,028 G. soja clones were assembled into 1053 FPC contigs covering approximately 547 Mbp of the G. max genome sequence. These physical maps for G. max and its undomesticated ancestor, G. soja, will serve as a framework for ordering sequence fragments, comparative genomics, cloning genes, and evolutionary analyses of legume genomes.

Ha, Jungmin; Abernathy, Brian; Nelson, William; Grant, David; Wu, Xiaolei; Nguyen, Henry T.; Stacey, Gary; Yu, Yeisoo; Wing, Rod A.; Shoemaker, Randy C.; Jackson, Scott A.



Molecular cloning and characterization of a new basic peroxidase cDNA from soybean hypocotyls infected with Phytophthora sojae f.sp. glycines.  


Differential display techniques were used to isolate cDNA clones corresponding to genes which were expressed in soybean hypocotyls by Phytophthora sojae f.sp. glycines infection. With a partial cDNA clone C20CI4 from the differential display PCR as a probe, a new basic peroxidase cDNA clone, designated GMIPER1, was isolated from a cDNA library of soybean hypocotyls infected with P. sojae f.sp. glycines. Sequence analysis revealed that the peroxidase clone encodes a mature protein of 35,813 Da with a putative signal peptide of 27 amino acids in its N-terminus. The amino acid sequence of the soybean peroxidase GMIPER1 is between 54-75% identical to other plant peroxidases including a soybean seed coat peroxidase. Southern blot analysis indicated that multiple copies of sequences related to GMIPER1 exist in the soybean genome. The mRNAs corresponding to the GMIPER1 cDNA accumulated predominantly in the soybean hypocotyls infected with the incompatible race of P. sojae f.sp. glycines, but were expressed at low levels in the compatible interaction. Soybean GMIPER1 mRNAs were not expressed in hypocotyls, leaves, stems, and roots of soybean seedlings. However, treatments with ethephon, salicylic acid or methyl jasmonate induced the accumulation of the GMIPER1 mRNAs in the different organs of soybean. These results suggest that the GMIPER1 gene encoding a putative pathogen-induced peroxidase may play an important role in induced resistance of soybean to P. sojae f.sp. glycines and in response to various external stresses. PMID:9856343

Yi, S Y; Hwang, B K



PsSAK1, a stress-activated MAP kinase of Phytophthora sojae, is required for zoospore viability and infection of soybean.  


Mitogen-activated protein kinase (MAPK) pathways are universal and evolutionarily conserved signal transduction modules in all eukaryotic cells. In this study, PsSAK1, which encodes a stress-activated MAPK of Phytophthora sojae, was identified. PsSAK1 is highly conserved in oomycetes, and it represents a novel group of MAPK due to its pleckstrin homology domain. Reverse-transcription polymerase chain reaction analysis showed that PsSAK1 expression was upregulated in zoospores and cysts and during early infection. In addition, its expression was induced by osmotic and oxidative stress mediated by NaCl and H(2)O(2), respectively. To elucidate the function, the expression of PsSAK1 was silenced using stable transformation of P. sojae. The silencing of PsSAK1 did not impair hyphal growth, sporulation, or oospore production but severely hindered zoospore development, in that the silenced strains showed quicker encystment and a lower germination ratio than the wild type. PsSAK1-silenced mutants produced much longer germ tubes and could not colonize either wounded or unwounded soybean leaves. Our results indicate that PsSAK1 is an important regulator of zoospore development and pathogenicity in P. sojae. PMID:20615113

Li, Aining; Wang, Yonglin; Tao, Kai; Dong, Suomeng; Huang, Qian; Dai, Tingting; Zheng, Xiaobo; Wang, Yuanchao



Genetic diversity of nodulating and non-nodulating rhizobia associated with wild soybean (Glycine soja Sieb. & Zucc.) in different ecoregions of China.  


A total of 99 bacterial isolates that originated from root nodules of Glycine soja were characterized with restriction analyses of amplified 16S ribosomal DNA and 16S-23S rDNA intergenic spacers (ITS), and sequence analyses of 16S rRNA, rpoB, atpD, recA and nodC genes. When tested for nodulation of G. soja, 72 of the isolates were effective symbionts, and these belonged to five species: Bradyrhizobium japonicum, Bradyrhizobium elkanii, Bradyrhizobium yuanmingense, Bradyrhizobium liaoningense and Sinorhizobium fredii. All of these, except some B. yuanmingense strains, also formed effective nodules on the domesticated soybean Glycine max. The remaining 27 isolates did not nodulate either host, but were identified as Rhizobium. Phylogeny nodC in the G. soja symbionts suggested that this symbiosis gene was mainly maintained by vertical gene transfer. Different nodC sublineages and rrs-ITS clusters reflected the geographic origins of isolates in this study. PMID:21303397

Wu, Li Juan; Wang, Hai Qing; Wang, En Tao; Chen, Wen Xin; Tian, Chang Fu



Survival and behavior of the insecticide-exposed predators Podisus nigrispinus and Supputius cincticeps (Heteroptera: Pentatomidae).  


Pentatomid stinkbugs are important predators of defoliating caterpillars in agricultural and forestry systems, and knowledge of the impact of insecticides on natural enemies is important information for integrated pest management (IPM) programs. Thus, we assessed the toxicity and behavioral sublethal response of the predators Podisus nigrispinus and Supputius cincticeps exposed to deltamethrin, methamidophos, spinosad and chlorantraniliprole, insecticides commonly used to control the velvetbean caterpillar (Anticarsia gemmatalis) in soybean crops. With the exception of deltamethrin for S. cincticeps, all insecticides showed higher acute toxicity to the prey than to these natural enemies providing effective control of A. gemmatalis. The recommended field concentration of deltamethrin, methamidophos and spinosad for controlling A. gemmatalis caused 100% mortality of P. nigrispinus and S. cincticeps nymphs. Chlorantraniliprole was the less toxic and the most selective insecticide to these predators resulting in mortalities of less than 10% when exposed to 10× the recommended field concentration for a period of 72h. Behavioral pattern changes in predators were found for all insecticides, especially methamidophos and spinosad, which exhibited irritability (i.e., avoidance after contact) to both predator species. However, insecticide repellence (i.e., avoidance without contact) was not observed in any of the insects tested. The lethal and sublethal effects of pesticides on natural enemies is of great importance for IPM, and our results indicate that substitution of pyrethroid and organophosphate insecticides at their field rates by chlorantraniliprole may be a key factor for the success of IPM programs of A. gemmatalis in soybeans. PMID:23880241

de Castro, A A; Corrêa, A S; Legaspi, J C; Guedes, R N C; Serrão, J E; Zanuncio, J C



Immunity of an Alternative Host Can Be Overcome by Higher Densities of Its Parasitoids Palmistichus elaeisis and Trichospilus diatraeae  

PubMed Central

Interactions of the parasitoids Palmistichus elaeisis Delvare & LaSalle and Trichospilus diatraeae Cherian & Margabandhu (Hymenoptera: Eulophidae) with its alternative host Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae) affect the success or failure of the mass production of these parasitoids for use in integrated pest management programs. The aim of this study was to evaluate changes in the cellular defense and encapsulation ability of A. gemmatalis pupae against P. elaeisis or T. diatraeae in adult parasitoid densities of 1, 3, 5, 7, 9, 11 or 13 parasitoids/pupae. We evaluated the total quantity of circulating hemocytes and the encapsulation rate versus density. Increasing parasitoid density reduced the total number of hemocytes in the hemolymph and the encapsulation rate by parasitized pupae. Furthermore, densities of P. elaeisis above 5 parasitoids/pupae caused higher reduction in total hemocyte numbers. The encapsulation rate fell with increasing parasitoid density. However, parasitic invasion by both species induced generally similar responses. The reduction in defensive capacity of A. gemmatalis is related to the adjustment of the density of these parasitoids to their development in this host. Thus, the role of the density of P. elaeisis or T. diatraeae by pupa is induced suppression of cellular defense and encapsulation of the host, even without them possesses a co-evolutionary history. Furthermore, these findings can predict the success of P. elaeisis and T. diatraeae in the control of insect pests through the use of immunology as a tool for evaluation of natural enemies.

Andrade, Gilberto Santos; Serrao, Jose Eduardo; Zanuncio, Jose Cola; Zanuncio, Teresinha Vinha; Leite, Germano Leao Demolin; Polanczyk, Ricardo Antonio



Hybridization between GM soybean (Glycine max (L.) Merr.) and wild soybean (Glycine soja Sieb. et Zucc.) under field conditions in Japan.  


Accumulation of information about natural hybridization between GM soybean (Glycine max) and wild soybean (Glycine soja) is required for risk assessment evaluation and to establish biosafety regulations in Japan. This is particularly important in areas where wild relatives of cultivated soybean are grown (i.e. East Asia including Japan). To collect information on temporal and spatial factors affecting variation in hybridization between wild and GM soybean, a two year hybridization experiment was established that included one wild soybean and five GM soybean cultivars with different maturity dates. Hybridization frequencies ranged from 0 to 0.097%. The maximum hybridization frequency (0.097%) was obtained from wild soybean crossed with GM soybean cv. AG6702RR, which were adjacently cultivated with wild soybean, with 25 hybrids out of 25 741 seedlings tested. Cultivar AG6702RR had the most synchronous flowering period with wild soybean. Ten hybrids out of 25 741 were produced by crossing with cv. AG5905RR, which had the second most synchronous flowering period with wild soybean. Most hybrids were found where GM and wild soybeans were adjacently cultivated, whereas only one hybrid was detected from wild soybean plants at 2 m, 4 m and 6 m from a pollen source (GM soybean). Differences in flowering phenology, isolation distance and presence of buffer plants accounted for half of the variation in hybridization frequency in this study. Temporal and spatial isolation will be effective strategies to minimize hybridization between GM and wild soybean. PMID:21122483

Mizuguti, Aki; Ohigashi, Kentaro; Yoshimura, Yasuyuki; Kaga, Akito; Kuroda, Yosuke; Matsuo, Kazuhito



Genetic Diversity and Population Structure: Implications for Conservation of Wild Soybean (Glycine soja Sieb. et Zucc) Based on Nuclear and Chloroplast Microsatellite Variation.  


Wild soybean (Glycine soja Sieb. et Zucc) is the most important germplasm resource for soybean breeding, and is currently subject to habitat loss, fragmentation and population decline. In order to develop successful conservation strategies, a total of 604 wild soybean accessions from 43 locations sampled across its range in China, Japan and Korea were analyzed using 20 nuclear (nSSRs) and five chloroplast microsatellite markers (cpSSRs) to reveal its genetic diversity and population structure. Relatively high nSSR diversity was found in wild soybean compared with other self-pollinated species, and the region of middle and lower reaches of Yangtze River (MDRY) was revealed to have the highest genetic diversity. However, cpSSRs suggested that Korea is a center of diversity. High genetic differentiation and low gene flow among populations were detected, which is consistent with the predominant self-pollination of wild soybean. Two main clusters were revealed by MCMC structure reconstruction and phylogenetic dendrogram, one formed by a group of populations from northwestern China (NWC) and north China (NC), and the other including northeastern China (NEC), Japan, Korea, MDRY, south China (SC) and southwestern China (SWC). Contrib analyses showed that southwestern China makes the greatest contribution to the total diversity and allelic richness, and is worthy of being given conservation priority. PMID:23202917

He, Shuilian; Wang, Yunsheng; Volis, Sergei; Li, Dezhu; Yi, Tingshuang



Biological safety of liposome-fullerene consisting of hydrogenated lecithin, glycine soja sterols, and fullerene-C60 upon photocytotoxicity and bacterial reverse mutagenicity.  


Various water-soluble derivatives of fullerene-C60 (C60) have been developed as detoxifiers for reactive oxygen species (ROS), whereas C60 incorporated in liposome (Lpsm) has not been reported yet. We prepared the liposome-fullerene (0.2% aqueous phase, Lpsm-Flln) which was composed of hydrogenated lecithin, glycine soja (soybean) sterols, and C60 in the weight ratio of 89.7:10:0.3, then examined the photocytotoxicity and bacterial reverse mutagenicity, as comparing with the Lpsm containing no C60. Photocytoxicity of Lpsm-Flln or Lpsm was examined using Balb/3T3 fibroblastic cells at graded doses of 0.49-1000 microg/mL under the condition of UVA- or sham-irradiation. The cells were irradiated with UVA (5 J/cm2, 320-400 nm, lambda max = 360 nm) at room temperature for 50 min. The resultant cell viability (% of control) did not decrease dose-dependently to 50% or less regardless of the UVA-irradiation. These results show that Lpsm-Flln or Lpsm does not possess photocytotoxicity to Balb/3T3 fibroblasts, and Lpsm-Flln may not exert a UVA-catalytic ROS-increasing action. A possibility for the reverse mutation by Lpsm-Flln or Lpsm was examined on four histidine-demanding strains of Salmonella typhimurium and a tryptophan-demanding strain of Escherichia coli. As for the dosages of Lpsm-Flln or Lpsm (313-5000 microg/plate), the dose-dependency of the number of reverse mutation colonies of each strain did not show a twice or more difference versus the negative control regardless of the metabolic activation, and, in contrast, marked differences for five positive controls (sodium azide, N-ethyl-N'-nitro-N-nitrosoguanidine, 2-nitrofluorene, 9-aminoacridine, and 2-aminoanthracene). The growth inhibition of bacterial strains and the deposition of Lpsm-Flln or Lpsm were not found. As a result, the bacterial reverse mutagenicity of Lpsm-Flln or Lpsm was judged to be negative under the conditions of this test. Thus, Lpsm-Flln and Lpsm may not give any significant biological toxic effects, such as photocytotoxicity and bacterial reverse mutagenicity. PMID:19482914

Kato, S; Aoshima, H; Saitoh, Y; Miwa, N



Effect of the insect growth regulator diflubenzuron on the predator Podisus nigrispinus (Heteroptera: Pentatomidae).  


Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) is a common natural predator of defoliating caterpillars in agricultural and forest systems. Insecticides acting as growth regulators of insect pests can indirectly affect their predators through consumption of contaminated prey. We examined the reproductive performance of P. nigrispinus fed on caterpillars of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae) reared on soybean leaves exposed to the chitin synthesis inhibitor, diflubenzuron. Caterpillars of A. gemmatalis were fed for 12 h with treated soybean leaves and offered to adults of the predator P. nigrispinus over five consecutive days. The fertility of P. nigrispinus was reduced when feeding on diflubenzuron treated caterpillars, especially at the beginning of the reproductive period, but recovered 3 weeks later. The effects of diflubenzuron ingestion on the life table parameters of P. nigrispinus included an increase in the period taken to double the population size, and reductions in the intrinsic rate of population increase, generation duration, and net reproductive rate. Diflubenzuron therefore had an indirect negative effect on the reproduction and the population dynamics of the non-target predator P. nigrispinus. Clearly, its use in integrated pest management requires further evaluation. PMID:21858643

Castro, Ancidériton A; Lacerda, Mabio C; Zanuncio, Teresinha V; de S Ramalho, Francisco; Polanczyk, Ricardo A; Serrão, José E; Zanuncio, José C



Enzymatic bioconversion of citrus hesperidin by Aspergillus sojae naringinase: enhanced solubility of hesperetin-7-O-glucoside with in vitro inhibition of human intestinal maltase, HMG-CoA reductase, and growth of Helicobacter pylori.  


Hesperetin-7-O-glucoside (Hes-7-G) was produced by the enzymatic conversion of hesperidin by Aspergillus sojae naringinase due to the removal of the terminal rhamnose. Extracts from orange juice and peel containing the hesperidin were so treated by this enzyme that the hesperidin could also be converted to Hes-7-G. The solubility of Hes-7-G in 10% ethanol was enhanced 55- and 88-fold over those of hesperidin and hesperetin, respectively, which may make Hes-7-G more bioavailable. Hes-7-G was 1.7- and 2.4-fold better than hesperidin and hesperetin, respectively, in the inhibition of human intestinal maltase. Hes-7-G was more potent by 2- and 4-fold than hesperidin in the inhibition of human HMG-CoA reductase. Additionally, Hes-7-G exhibited more effective inhibition of the growth of Helicobacter pylori than hesperetin, while its effectiveness was similar to that of hesperidin. Therefore, the results suggest that bioconverted Hes-7-G is more effective and bioavailable than hesperidin, as it has enhanced inhibitory and solubility properties. PMID:22980799

Lee, Young-Su; Huh, Ji-Young; Nam, So-Hyun; Moon, Sung-Kwon; Lee, Soo-Bok



Characterization of a novel Cry9Bb delta-endotoxin from Bacillus thuringiensis.  


The Brazilian Bacillus thuringiensis serovar japonensis strain S725 was selected for its toxicity to the velvetbean caterpillar, Anticarsia gemmatalis. This strain produces spherical crystals harbouring a major protein of about 130 kDa which yields fragments of between 50 and 70 kDa upon trypsin activation. The protein showed a high level of identity and immunoafinity to the Cry9 class of delta-endotoxins. The cloned cry9-like gene sequence contains a 3492 bp ORF, which encodes a polypeptide of 1163 amino acids, with a predicted molecular mass of 131.4 kDa. The deduced amino acid sequence is unique and shows 73% identity to Cry9Ba, 64% identity to Cry9Ea, 63% identity to Cry9Da, and 59% identity to Cry9Ca proteins. The novel delta-endotoxin was assigned to a new subclass, Cry9Bb, by the Bt Toxin Nomenclature Committee. The Cry9Bb protein was expressed in an acrystalliferous Bt strain, and exhibited activity against the tobacco hornworm, Manduca sexta, and the velvetbean caterpillar, A. gemmatalis. The biological effect of an amino acid residue change, A84P, was investigated. The LC(50) for the Cry9Bb crystals against M. sexta neonate larvae was 6.84 microg/cm(2), while the LC(50) for the mutant's Cry9Bb crystals was 0.78 microg/cm(2). PCR screening revealed that in addition to cry9Bb, Bt strain S725 also contains cry1I and vip3 genes. Transcription analysis, using RT-PCR, showed that the cry1I gene was transcribed at T(2) and T(5) stages of sporulation. PMID:18456277

Silva-Werneck, J O; Ellar, D J



Field evaluation of soybean engineered with a synthetic cry1Ac transgene for resistance to corn earworm, soybean looper, velvetbean caterpillar (Lepidoptera: Noctuidae), and lesser cornstalk borer (Lepidoptera: Pyralidae).  


A transgenic line of the soybean 'Jack', Glycine max (L.) Merrill, expressing a synthetic cry1Ac gene from Bacillus thuringiensis variety kurstaki (Jack-Bt), was evaluated for resistance to four lepidopteran pests in the field. Jack-Bt and genotypes serving as susceptible and resistant controls were planted in field cages and artificially infested with larvae of corn earworm, Helicoverpa zea (Boddie), and velvetbean caterpillar, Anticarsia gemmatalis (Hübner), in 1996, 1997, and 1998, and also with soybean looper, Pseudoplusia includens (Walker), in 1996. Susceptible controls included Jack (1996-1998), 'Cobb' (1996), and Jack-HPH (1996). GatIR 81-296 was used as the resistant control in all 3 yr. Compared with untransformed Jack, Jack-Bt showed three to five times less defoliation from corn earworm and eight to nine times less damage from velvetbean caterpillar. Defoliation of GatIR 81-296 was intermediate between that of Jack and Jack-Bt for corn earworm, and similar to that of Jack for velveltbean caterpillar. Jack-Bt exhibited significant, but lower resistance to soybean looper. Jack-Bt also showed four times greater resistance than Jack to natural infestations of lesser cornstalk borer, Elasmopalpus lignosellus (Zeller), in conventional field plots at two locations in 1998. Data from these experiments suggest that expression of this cry1Ac construct in soybean should provide adequate levels of resistance to several lepidopteran pests under field conditions. PMID:10902306

Walker, D R; All, J N; McPherson, R M; Boerma, H R; Parrott, W A



Perception of solar UVB radiation by phytophagous insects: Behavioral responses and ecosystem implications  

PubMed Central

Most of our present knowledge about the impacts of solar UVB radiation on terrestrial ecosystems comes from studies with plants. Recently, the effects of UVB on the growth and survival of consumer species have begun to receive attention, but very little is known about UVB impacts on animal behavior. Here we report that manipulations of the flux of solar UVB received by field-grown soybean crops had large and consistent effects on the density of the thrips (Caliothrips phaseoli, Thysanoptera: Thripidae) populations that invaded the canopies, as well as on the amount of leaf damage caused by the insects. Solar UVB strongly reduced thrips herbivory. Thrips not only preferred leaves from plants that were not exposed to solar UVB over leaves from UVB-exposed plants in laboratory and field choice experiments, but they also appeared to directly sense and avoid exposure to solar UVB. Additional choice experiments showed that soybean leaf consumption by the late-season soybean worm Anticarsia gemmatalis (Lepidoptera: Noctuidae) was much less intense in leaves with even slight symptoms of an early thrips attack than in undamaged leaves. These experiments suggest that phytophagous insects can present direct and indirect behavioral responses to solar UVB. The indirect responses are mediated by changes in the plant host that are induced by UVB and, possibly, by other insects whose behavior is affected by UVB.

Mazza, Carlos A.; Zavala, Jorge; Scopel, Ana L.; Ballare, Carlos L.



Genetic transformation, recovery, and characterization of fertile soybean transgenic for a synthetic Bacillus thuringiensis cryIAc gene.  

PubMed Central

Somatic embryos of jack, a Glycine max (L.) Merrill cultivar, were transformed using microprojectile bombardment with a synthetic Bacillus thuringiensis insecticidal crystal protein gene (Bt cryIAc) driven by the 35S promoter and linked to the HPH gene. Approximately 10 g of tissue was bombarded, and three transgenic lines were selected on hygromycin-containing media and converted into plants. The recovered lines contained the HPH gene, but the Bt gene was lost in one line. The plasmid was rearranged in the second line, and the third line had two copies, one of which was rear-ranged. The CryIAc protein accumulated up to 46 ng mg-1 extractable protein. In detached-leaf bioassays, plants with an intact copy of the Bt gene, and to a lesser extent those with the rearranged copy, were protected from damage from corn earworm (Helicoverpa zea), soybean looper (Pseudoplusia includens), tobacco budworm (Heliothis virescens), and velvetbean caterpillar (Anticarsia gemmatalis). Corn earworm produced less than 3% defoliation on transgenic plants, compared with 20% on the lepidopteran-resistant breeding line GatIR81-296, and more than 40% on susceptible cultivars. Unlike previous reports of soybean transformation using this technique, all plants were fertile. To our knowledge, this is the first report of a soybean transgenic for a highly expressed insecticidal gene.

Stewart, C N; Adang, M J; All, J N; Boerma, H R; Cardineau, G; Tucker, D; Parrott, W A



Characterization of a nucleopolyhedrovirus from the black cutworm, agrotis ipsilon (Lepidoptera: noctuidae)  


The black cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), is a serious localized pest of vegetable and field crops. We have characterized a newly discovered baculovirus, the Agrotis ipsilon multicapsid nucleopolyhedrovirus (AgipMNPV), that was isolated from A. ipsilon in Illinois. Restriction enzyme fragment profiles of AgipMNPV DNA were distinct from those of previously described nucleopolyhedroviruses. Electron microscopy of AgipMNPV-infected tissues indicated that nucleocapsids of this virus are multiply enveloped. A. ipsilon was highly susceptible to infection by AgipMNPV and significantly more susceptible to infection by AgipMNPV than by Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV). Host range studies showed that Heliothis virescens and Helicoverpa zea were moderately susceptible to infection; Pseudaletia unipuncta and Spodoptera frugiperda were only partially susceptible, and Anticarsia gemmatalis, Spodoptera exigua, Trichoplusia ni, and Ostrinia nubilalis were not susceptible to infection by AgipMNPV. Because of its high virulence, AgipMNPV has potential as an alternative to chemical insecticides for control of A. ipsilon. Copyright 1999 Academic Press. PMID:10534417

Boughton; Harrison; Lewis; Bonning



An Amino Acid Substitution Inhibits Specialist Herbivore Production of an Antagonist Effector and Recovers Insect-Induced Plant Defenses1[W][OA  

PubMed Central

Plants respond to insect herbivory through the production of biochemicals that function as either direct defenses or indirect defenses via the attraction of natural enemies. While attack by closely related insect pests can result in distinctive levels of induced plant defenses, precise biochemical mechanisms responsible for differing responses remain largely unknown. Cowpea (Vigna unguiculata) responds to Fall armyworm (Spodoptera frugiperda) herbivory through the detection of fragments of chloroplastic ATP synthase ?-subunit proteins, termed inceptin-related peptides, present in larval oral secretions (OS). In contrast to generalists like Fall armyworm, OS of the legume-specializing velvetbean caterpillar (VBC; Anticarsia gemmatalis) do not elicit ethylene production and demonstrate significantly lower induced volatile emission in direct herbivory comparisons. Unlike all other Lepidoptera OS examined, which preferentially contain inceptin (Vu-In; +ICDINGVCVDA?), VBC OS contain predominantly a C-terminal truncated peptide, Vu-In?A (+ICDINGVCVD?). Vu-In?A is both inactive and functions as a potent naturally occurring antagonist of Vu-In-induced responses. To block antagonist production, amino acid substitutions at the C terminus were screened for differences in VBC gut proteolysis. A valine-substituted peptide (Vu-In?V; +ICDINGVCVDV?) retaining full elicitor activity was found to accumulate in VBC OS. Compared with the native polypeptide, VBC that previously ingested 500 pmol of the valine-modified chloroplastic ATP synthase ?-subunit precursor elicited significantly stronger plant responses in herbivory assays. We demonstrate that a specialist herbivore minimizes the activation of defenses by converting an elicitor into an antagonist effector and identify an amino acid substitution that recovers these induced plant defenses to a level observed with generalist herbivores.

Schmelz, Eric A.; Huffaker, Alisa; Carroll, Mark J.; Alborn, Hans T.; Ali, Jared G.; Teal, Peter E.A.




Microsoft Academic Search

This study aimed to apply an exploratory research to observe the consumption behavior of petit suisse cheese between 120 students and employees from the Federal University of Technology at the Medianeira campus. Results pointed out that 80% were women and 58,33% presented average age betweem 20-40 years. This study pointed out that 53,33% consumed soy products one to three times

Elizete Neuza Brach; Henry Charles; Albert David; Naidoo Terroso


Phyto-œstrogènes du soja : problèmes posés chez le nourrisson allergique au lait de vache et consommant des formules à base de soja  

Microsoft Academic Search

This work presents isoflavones levels in soy-based infant formulas commercialised in France. These compounds are known to be both estrogenic and anti-gonadotrophic compounds. A comparison is made with soy based food supplements for estrogen deprived menopausal women. This work gives the latest scientific data on the effects of isoflavones in humans and presents the actual controversy on the effect of

Catherine Bennetau-Pelissero; P. Sauvant; G. Peltre; P. Auriol; Anna Rocca; Fabienne Rancé



Bradyrhizobium elkanii lipo-oligosaccharide signals induce complete nodule structures on Glycine soja Siebold et Zucc  

Microsoft Academic Search

The (Brady)rhizobium nodulation gene products synthesize lipo-oligosaccharide (LOS) signal molecules that induce nodule primordia on legume roots. Previously, nodule primordia had been induced by LOS on several legumes, but the induction of a complete nodule structure by LOS had only been shown on alfalfa. Since alfalfa can spontaneously form nodules at a low frequency, it has been unclear whether LOS

Thomas J. W. Stokkermans; N. Kent Peters



Qualidade fisiológica de sementes de milho, feijão, soja e alface na presença de extrato de tiririca  

Microsoft Academic Search

RESUMO - A espécie Cyperus rotundus (tiririca) é perene e de difícil controle. Os órgãos subterrâneos dessa ciperácea produzem inibidores capazes de interferir na germinação e no crescimento de plântulas e de plantas de várias espécies, fenômeno chamado de alelopatia. A inibição na germinação de sementes pode estar associada à interferência de substâncias alelopáticas na atividade de enzimas chaves no

Fabiana Rezende Muniz; Maria das Graças Cardoso; Édila Vilela Resende Von Pinho; Michelle Vilela




Microsoft Academic Search

UTILIZATION OF FULL-FAT SOYBEAN WITHOUT KUNITZ FACTOR IN BROWN-EGG LAYER DIETS. Two hundred and eighty brown-egg layers (56\\/treatment) were individually alloted in cages and subjected to a completely randomized design. The birds were fed increasing dietary levels (0; 5; 10; 15; and 20%) of low Kunitz raw soybean during 84 days. Egg shell quality and yolk color were not affected

Paulo R; Rogério MAGGIONI


Ectopic Expression of a WRKY Homolog from Glycine soja Alters Flowering Time in Arabidopsis.  


Flowering is a critical event in the life cycle of plants; the WRKY-type transcription factors are reported to be involved in many developmental processes sunch as trichome development and epicuticular wax loading, but whether they are involved in flowering time regulation is still unknown. Within this study, we provide clear evidence that GsWRKY20, a member of WRKY gene family from wild soybean, is involved in controlling plant flowering time. Expression of GsWRKY20 was abundant in the shoot tips and inflorescence meristems of wild soybean. Phenotypic analysis showed that GsWRKY20 over-expression lines flowered earlier than the wild-type plants under all conditions: long-day and short-day photoperiods, vernalization, or exogenous GA3 application, indicating that GsWRKY20 may mainly be involved in an autonomous flowering pathway. Further analyses by qRT-PCR and microarray suggests that GsWRKY20 accelerating plant flowering might primarily be through the regulation of flowering-related genes (i.e., FLC, FT, SOC1 and CO) and floral meristem identity genes (i.e., AP1, SEP3, AP3, PI and AG). Our results provide the evidence demonstrating the effectiveness of manipulating GsWRKY20 for altering plant flowering time. PMID:23991184

Luo, Xiao; Sun, Xiaoli; Liu, Baohui; Zhu, Dan; Bai, Xi; Cai, Hua; Ji, Wei; Cao, Lei; Wu, Jing; Wang, Mingchao; Ding, Xiaodong; Zhu, Yanming




Microsoft Academic Search

The transfer of seed storage material from the cotiledons to the embryo axys of soybean seeds (Glycine max (L.) Merrill) was studied. Two lots of cultivar Bragg with germination above 80% and differing in vigor levels, as assessed by the accelerated aging test were used. The initial seed quality was evaluated by moisture content, weight of 1,000 seeds, germination and



Filières soja OGM et non OGM : à quelles conditions la coexistence est-elle possible ?  

Microsoft Academic Search

L'Union européenne a mis en place une réglementation dont l'objectif est d’encadrer les disséminations volontaires d’organismes génétiquement modifiés (OGM) dans l’environnement. Elle laisse aux Etats membres la possibilité de fixer des mesures de coexistence des cultures OGM et non OGM pour satisfaire au libre choix des producteurs agricoles et des consommateurs face aux produits issus, ou non, d'organismes génétiquement modifiés.

Romain Bourgier; Raùl Green; Marianne Le Bail; Louis-Georges Soler



Identification and Functional Characterization of the Soybean GmaPPO12 Promoter Conferring Phytophthora sojae Induced Expression  

PubMed Central

Identification of pathogen-inducible promoters largely lags behind cloning of the genes for disease resistance. Here, we cloned the soybean GmaPPO12 gene and found that it was rapidly and strongly induced by Phytophthorasojae infection. Computational analysis revealed that its promoter contained many known cis-elements, including several defense related transcriptional factor-binding boxes. We showed that the promoter could mediate induction of GUS expression upon infection in both transient expression assays in Nicotianabenthamiana and stable transgenic soybean hairy roots. Importantly, we demonstrated that pathogen-induced expression of the GmaPPO12 promoter was higher than that of the soybean GmaPR1a promoter. A progressive 5’ and 3’ deletion analysis revealed two fragments that were essential for promoter activity. Thus, the cloned promoter could be used in transgenic plants to enhance resistance to phytophthora pathogens, and the identified fragment could serve as a candidate to produce synthetic pathogen-induced promoters.

Chai, Chunyue; Lin, Yanling; Shen, Danyu; Wu, Yuren; Li, Hongjuan; Dou, Daolong




Microsoft Academic Search

This study was conducted to determine the effect of planting date on soybean seed quality. The cultivars evaluated were Cristalina, IAC-2, Tropical, Doko, and IAC-8. The sowing dates were stablished between October 29, 1983 and January 7, 1984 (10\\/29, 11\\/11, 11\\/25, 12\\/10, 12\\/24 and 01\\/7), at Lucas do Rio Verde in Mato Grosso State, Brazil. In the plantings carried out



Genetic Variation of Wild Soybean Glycine soja Sieb. et Zucc. in the Far East Region of the Russian Federation  

Microsoft Academic Search

Polymorphism of RAPD markers was analyzed in the wild soybean populations from the Far East region of Russia. The level of RAPD marker polymorphism was significantly higher in the wild than in the cultivated soybean. The results obtained suggest active development of genetically different groups of wild soybean. Geographically isolated subpopulations showed maximum distance from the main population of wild

A. M. Seitova; A. N. Ignatov; T. P. Suprunova; I. L. Tsvetkov; E. V. Deineko; D. B. Dorokhov; V. K. Shumnyi; K. G. Skryabin



Preferential nodulation of Glycine max, Glycine soja and Macroptilium atropurpureum by two Bradyrhizobium species japonicum and elkanii  

Microsoft Academic Search

Soybean bradyrhizobia, Bradyrhizobium japonicum and Bradyrhizobium elkanii differ in various traits such as DNA fingerprint, rhizobitoxine production, indole-3-acetic acid production and uptake of hydrogenase. In this communication, we investigated whether the differences between both species extend to host preference in multistrain environments. Nodule occupancy of B.japonicum and B. elkanii significantly depended on host plants. B. japonicum and B. elkanii preferentially

Kiwamu Minamisawa; Shingo Onodera; Yasuaki Tanimura; Naoki Kobayashi; Ken-Ichi Yuhashi; Masatsugu Kubota



Consumo de energia eletrica na moagem de sorgo e soja utilizados na alimentacao de frangos. (Electric energy consumption in soybean and sorghum milling used in chickens rations).  

National Technical Information Service (NTIS)

This research aims the evaluation of the performance of milling machines, considering the electric energy consumption to obtain rations of different granulometry and nutritive quality analysis of this rations in relation to chicken growth. 50 refs, 60 tab...

M. D. Mendonca




Microsoft Academic Search

The aim of the study was to evaluate the effectiveness of a combination of personal protective equipments (PPE) for the tractor operator applying herbicides on soybean and peanut crops with a tractor mounted boom sprayer and the safetyness of these work conditions. The application of the herbicides were in pre-planting soil-incorporated (ppi), pre-emergence (pre) and post-emergence (post) treatment, with volumes



Uso DA Expansao Direta Para Estimar Areas de Soja E Milho Atraves de Dados Multiespectrais E Temporais Do Landsat/TM (Use of Direct Expansion for Estimation of Soybean and Corn Areas Through Multispectral and Temporal Landsat/TM Data).  

National Technical Information Service (NTIS)

The contribution of LANDSAT/TM data in estimating soybean and corn areas was analyzed through a direct expansion method by sampling techniques. The study was carried out in the test site of Distrito Federal (5,814 sq km), covered by LANDSAT/TM (row/path 2...

M. Alvesmoreira



Evolução clínica e laboratorial de crianças com alergia a leite de vaca e ingestão de bebida à base de soja Clinical and laboratorial follow-up of children with cow's milk allergy (CMA) using a soy based beverage  

Microsoft Academic Search

Objective: To evaluate the use of a soy beverage in infants with cow's milk allergy (CMA), considering all nutritional aspects and the development of soy allergy. Methods: Twelve CMA patients with 1 to 4 years old received soy beverage and were subjected to clinical, labo- ratory and nutritional assessment during 12 months. Soy formula samples were evaluated by ELISA technique

Ana Paula; B. M. Castro; Cristina Miuki; A. Jacob; Gabriela Ackel Corradi; Dulcineia Abdalla; Renata F. F. Gonçalves; Fabia Thais; L. Rocha; Antonio Carlos Pastorino


NODULAÇÃO E CRESCIMENTO DE VARIEDADES DE SOJA RR SOB APLICAÇÃO DE GLYPHOSATE, FLUAZIFOP-P-BUTYL E FOMESAFEN1 GR Glycine max Nodulation and Growth Under Glyphosate, Fluazifop-p-Butyl and Fomesafen Aplication  

Microsoft Academic Search

The objective of this work was to evaluate the effects of the herbicides glyphosate, fluazifop-p-butyl and fomesafen, applied by different methods, on nodulation and initial growth of two glyphosate-resistant (GR) soybean varieties. Treatments were arranged in a 6 x 2 factorial, with six herbicide application methods (no-herbicide check, fomesafen\\/fomesafen sequential application, fomesafen\\/(fomesafen+fluazifop-p-butyl sequential application), fluazifop-p-butyl single application, glyphosate\\/glyphosate sequential application



Molho cremoso à base de extrato de soja: estabilidade, propriedades reológicas, valor nutricional e aceitabilidade do consumidor Soy - based low fat emulsion: stability, rheology, nutritional value and consumer acceptance  

Microsoft Academic Search

The salad dressing preparation, which replaces for replacing the mayonnaise, uses utilizes emulsifiers and thickeners related or not to the soy protein in order to reduce the egg and oil quantity. In the The present work, used homogenized whole soymilk was used to prepare a creamy low-fat emulsion to replace traditional mayonnaise. The whole soymilk was elaborated with 6-12% of

Denise Carvalho; Pereira CAMPOS; Rosemar ANTONIASSI; Rosires DELIZA; Sidinéa Cordeiro de FREITAS; Ilana FELBERG




Technology Transfer Automated Retrieval System (TEKTRAN)

No white-flowered accession exists among the more than 1,100 Glycine. soja accessions in the USDA Soybean Germplasm Collection, although one-third of the Glycine max accessions are white-flowered. One white-flowered plant was found in G. soja accession PI 424008A growing in Stoneville, Mississippi i...


Constructing the Self in/as Thirdspace: New Potentials for Identity Exploration in the Composition Classroom  

ERIC Educational Resources Information Center

|In this article the author introduces a concept she calls "Thirdspace identity construction," which instructors can use to understand what happens in students' texts when such ever-open possibilities for identity exploration are allowed. This concept borrows from the work of critical geographer Edward Soja. Soja's "Thirdspace" represents a…

Lauer, Claire



QTL affecting fitness of hybrids between wild and cultivated soybeans in experimental fields  

PubMed Central

The objective of this study was to identify quantitative trait loci (QTL) affecting fitness of hybrids between wild soybean (Glycine soja) and cultivated soybean (Glycine max). Seed dormancy and seed number, both of which are important for fitness, were evaluated by testing artificial hybrids of G. soja × G. max in a multiple-site field trial. Generally, the fitness of the F1 hybrids and hybrid derivatives from self-pollination was lower than that of G. soja due to loss of seed dormancy, whereas the fitness of hybrid derivatives with higher proportions of G. soja genetic background was comparable with that of G. soja. These differences were genetically dissected into QTL for each population. Three QTLs for seed dormancy and one QTL for total seed number were detected in the F2 progenies of two diverse cross combinations. At those four QTLs, the G. max alleles reduced seed number and severely reduced seed survival during the winter, suggesting that major genes acquired during soybean adaptation to cultivation have a selective disadvantage in natural habitats. In progenies with a higher proportion of G. soja genetic background, the genetic effects of the G. max alleles were not expressed as phenotypes because the G. soja alleles were dominant over the G. max alleles. Considering the highly inbreeding nature of these species, most hybrid derivatives would disappear quickly in early self-pollinating generations in natural habitats because of the low fitness of plants carrying G. max alleles.

Kuroda, Yosuke; Kaga, Akito; Tomooka, Norihiko; Yano, Hiroshi; Takada, Yoshitake; Kato, Shin; Vaughan, Duncan



Antifungal activity of borrelidin produced by a Streptomyces strain isolated from soybean.  


In this study, an endophytic Streptomyces sp. neau-D50 with strong antifungal activity against Phytophthora sojae was isolated from healthy soybean root, using an in vitro screening technique. A bioactivity-guided approach was then employed to isolate and determine the chemical identity of bioactive constituents with antifungal activity from strain neau-D50. The structure of the antifungal metabolite was elucidated as borrelidin on the basis of spectral analysis. To our knowledge, this is the first report that borrelidin has strong antifungal activity against dominant race 1 of P. sojae with EC(50) and EC(95) of 0.0056 and 0.026 mg/L, respectively. The values were respectively 62.5- and 262.3-fold lower than those of the commercial fungicide metalaxyl, which has been used to treat soybean seed for the control of P. sojae . The in situ bioassays demonstrated that borrelidin at 10 mg/L reduced P. sojae race 1 lesions on soybean seedlings by 94.72% without affecting root growth. Thus, borrelidin might be a promising candidate for new antifungal agents against P. sojae. PMID:22242825

Liu, Chong-Xi; Zhang, Ji; Wang, Xiang-Jing; Qian, Ping-Ting; Wang, Ji-Dong; Gao, Ya-Mei; Yan, Yi-Jun; Zhang, Shu-Zhen; Xu, Peng-Fei; Li, Wen-Bin; Xiang, Wen-Sheng



The effect of dietary gamma-tocopherol on serum tocopherols in formulated infants.  


The levels of alpha-, beta- and gamma-tocopherol were determined in two formulas (Nutramigen and Soja-semp) and in 20 infants fed either of these formulas. The gamma-tocopherol concentration in Nutramigen was 22.8 +/- 1.9 mumol/l; and that in Soja-semp 3.7 +/- 0.5 mumol/l. This difference was reflected in the serum tocopherols of the infants, since in the 10 infants fed Nutramigen, gamma-tocopherol accounted for 16.7 +/- 7.9% of the total tocopherols compared to 4.2 +/- 1.8% in the 10 infants fed Soja-semp. The study shows that gamma-tocopherol accumulates in the serum of infants fed large amounts of it. The serum gamma-tocopherol level should therefore be taken into account, when estimating the vitamin E status in humans, especially when the intake of gamma-tocopherol is high. PMID:7246124

Jansson, L; Holmberg, L; Jakobsson, I



Host-Pathogen Interactions  

PubMed Central

The glucan elicitor isolated from the mycelial walls of Phytophthora megasperma var. sojae, the fungus which causes stem and root rot in soybeans, stimulates the activity of phenylalanine ammonia-lyase and the accumulation of glyceollin in suspension-cultured soybean cells. Nigeran, a commercially available fungal wall glucan, was the only other compound tested which has any activity in this system. Glyceollin is a phenylpropanoid-derived phytoalexin which is toxic to P. megasperma var. sojae. Evidence is presented to support the hypothesis that the action of elicitors in stimulating phytoalexin synthesis is not species or variety specific but, rather, is part of a general defensive response of plants.

Ebel, Jurgen; Ayers, Arthur R.; Albersheim, Peter



Amplified fragment length polymorphism (AFLP) in soybean: species diversity, inheritance, and near-isogenic line analysis  

Microsoft Academic Search

Amplified fragment length polymorphism (AFLP) analysis is a PCR-based technique capable of detecting more than 50 independent loci in a single PCR reaction. The objectives of the present study were to: (1) assess the extent of AFLP variation in cultivated (Gycine max L. Merr.) and wild soybean (G. soja Siebold & Zucc.), (2) determine genetic relationships among soybean accessions using

P. J. Maughan; M. A. Sagllai Maroof; G. R. Buss; G. M. Huestis




Technology Transfer Automated Retrieval System (TEKTRAN)

Two gene pairs are known to affect leaflet shape in soybean (Glycine max [L.] Merr.), but only two categories of leaflet shape are routinely used to characterize soybean germplasm. Very little information has been published about leaflet shape and size for wild soybean (Glycine soja Sieb. & Zucc.) a...


Nutrition rehabilitation of undernourished children utilizing Spiruline and Misola  

Microsoft Academic Search

BACKGROUND: Malnutrition constitutes a public health problem throughout the world and particularly in developing countries. AIMS: The objective of the study is to assess the impact of an elementary integrator composed of Spiruline (Spirulina platensis) and Misola (millet, soja, peanut) produced at the Centre Medical St Camille (CMSC) of Ouagadougou, Burkina Faso, on the nutritional status of undernourished children. MATERIALS

Jacques Simpore; Fatoumata Kabore; Frederic Zongo; Deleli Dansou; Augustin Bere; Salvatore Pignatelli; Daniela M Biondi; Giuseppe Ruberto; Salvatore Musumeci



Footprints of evolution: the dynamics of effector genes in the Phytophthora genome  

Microsoft Academic Search

Phytophthora is a genus comprised of over 65 destructive plant pathogenic species that cause severe damages in agriculture, forestry and natural habitats. Economically important pathogens are Phytophthora infestans (causing potato late blight) and Phytophthora sojae (causing soybean root and stem rot). A newly discovered species, Phytophthora ramorum is destroying oak trees along the west-coast of the USA by causing the

R. H. Y. Jiang



Cryptic Sexuality in Aspergillus parasiticus and A. flavus  

Technology Transfer Automated Retrieval System (TEKTRAN)

Ascomycetous fungi of the genus Aspergillus comprise a wide variety of species of biotechnological importance (e.g. A. sojae, A. oryzae, A. niger) as well as pathogens and toxin producers (e.g. A. flavus, A. parasiticus, A. fumigatus, A. nidulans). With the exception of A. nidulans, which is a homot...


Phytophthora genomics: the plant destroyers' genome decoded  

Microsoft Academic Search

The year 2004 was an exciting one for the Phytophthora research community. The United States Department of Energy Joint Genome Institute (JGI) completed the draft genome sequence of two Phytophthora species, Phytophthora sojae and Phytophthora ramorum. In August of that year over 50 people gathered at JGI in Walnut Creek, California, for an annotation jamboree and searched for the secrets

Francine Govers; Mark Gijzen




Technology Transfer Automated Retrieval System (TEKTRAN)

Draft genome sequences of the soybean pathogen Phytophthora sojae and the sudden oak death pathogen Phytophthora ramorum have been determined. Oomycetes such as these Phytophthora species share the kingdom Stramenopiles with photosynthetic algae such as diatoms, and the Phytophthora sequences sugges...


American Ethni\\/Cities: Critical Geography, Subject Formation, and the Urban Representations of Abraham Cahan, Richard Wright, and James Baldwin  

Microsoft Academic Search

By drawing upon aspects of critical geography to explore three writers' representations of urban space and subject formation, American Ethni\\/Cities develops and advocates for a new methodological approach to the study of literature. Predicated on theories devised by Henri Lefebvre, David Harvey, Edward Soja, Gil Valentine and other geographically-minded thinkers, this spatially conscious literary practice has the potential to enhance

Joshua Scott Stone



Fermentação da proteína de seis alimentos por microrganismos ruminais, incubados puros ou com monensina ou rumensin®  

Microsoft Academic Search

RESUMO - Avaliaram-se os efeitos da fermentação in vitro de seis alimentos: fubá de milho (FM), farelo de soja (FS), farelo de trigo (FT), sorgo (SO), glúten de milho (GM) e uréia (UR), incubados puros ou com o antibiótico monensina, esse na forma pur a para análise (Monensina) ou comercial (Rumensin®). O experimento constituiu-se de 18 unidades experimentais (três alimentos

Natália Guarino Souza Barbosa; Rogério de Paula Lana; Antônio Bento Mâncio; Arnaldo Chaer Borges; Cesar de Queiroz; Juliana Silva Oliveira




Technology Transfer Automated Retrieval System (TEKTRAN)

A combined proteomics approach was applied for the separation, identification, and comparison of two major storage proteins, '-conglycinin and glycinin in wild (Glycine soja) and cultivated (Glycine max) soybean seeds. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) with three immobili...



Microsoft Academic Search

Plant-associated bacteria that live inside plant tissues without causing any harm to plants are defined as endophytic bacteria. The present investigation was carried out to analyse the phenotypic and genotypic diversity in the bacterial endophytes of two species of soybean viz. Glycine max and G. soja . A total of 65 bacterial endophytes were isolated from three tissues: stem, root

Pham Quang Hung; K. Annapurna


Effect of root temperature, plant age, frequency and duration of flooding and inoculum placement and concentration on susceptibility of asparagus to Phytophthora rot  

Microsoft Academic Search

The conditions and recommended procedure to maximise disease severity for greenhouse screening of asparagus seedlings for resistance to Phytophthora rot are described. The optimum temperature for disease development on asparagus seedlings inoculated with Phytophthora megasperma var. sojae was 15°C and 18–24°C for P. cryptogea. The same temperatures were also optimal for indirect germination of sporangia of the two species. Young

Peter G. Falloon; R. G. Grogan



An Integrated Genetic Linkage Map of the Soybean Genome  

Microsoft Academic Search

highly informative DNA markers evenly spaced throughout the genome. A number of molecular genetic maps of the soybean (Glycine max Using a mapping population derived from an inter- (L.) Merr.) have been developed over the past 10 yr. These maps are specific G. max 3 G. soja cross, Shoemaker and Olson primarily based on restriction fragment length polymorphism (RFLP) (1993)

P. B. Cregan; T. Jarvik; A. L. Bush; R. C. Shoemaker; K. G. Lark; A. L. Kahler; N. Kaya; T. T. VanToai; D. G. Lohnes; J. Chung; J. E. Specht



La production de viandes végétales  

Microsoft Academic Search

[eng] Cereal and animal proteins share a certain past (the Experience of Beccari in 1728, Liebig's Hydrolysats, etc). However most new synthetic meats appearing in the U.S. are not derived from cereals but from a vegetable plant : soja. . Huge milling groups have created and put on the market these new

Jean Bure



Changing Course: Locating Third Space in a College Classroom  

ERIC Educational Resources Information Center

|The author reflects on her experience teaching undergraduate children's literature over several semesters and the impact of her action research study on her teaching. Using theoretical concepts of Third Space (Bhabha, 1994; Soja, 1996), the author examines the evolution of her philosophy of education and classroom practice. The author determines…

Johnston, Jennifer A.




Technology Transfer Automated Retrieval System (TEKTRAN)

Draft genome sequences of the soybean pathogen Phytophthora sojae and the sudden oak death pathogen Phytophthora ramorum have been determined to depths of 9x and 7.7x, respectively. Oomycetes such as these Phytophthora species share the kingdom Stramenopiles with photosynthetic algae such as diatoms...


Effect of salt stress on seed germination and seedling growth of three salinity plants.  


Seeds of three halophyte species included Limonium sinense Kuntze, Glycine soja sieb. and Sorghum sudanense Stapf. were used to investigate the effects of different salinity concentrations on their germination percentage, germination energy, germination index, relative germination rate, relative salt-injury rate, radicle length and hypocotyl length. Results showed that seeds of G. soja can germinate well and rapidly at lower level of salt (< 200 mmol L(-1)), 50 mmol L(-1) salt concentrations is better to S. sudanense, suitable salt concentrations was beneficial to germinate of S. sudanense seeds. Seeds of L. sinense and S. sudanense can germinate at higher level of salt (400 mmol L(-1)). Three plant seeds had different physiological mechanism for germination under salt stress, radicle hypocotyl ratio of L. sinense and S. sudanense increased with increasing salt suggested that they had the higher tolerance in shoot growth under NaCl stress, which is perhaps the reason for their wide utilisation for saline soil rehabilitation. PMID:18819537

Li, Yan



Tryptophan Biosynthesis in Stramenopiles: Eukaryotic Winners in the Diatom Complex Chloroplast  

Microsoft Academic Search

Tryptophan is an essential amino acid that, in eukaryotes, is synthesized either in the plastids of photoautotrophs or in\\u000a the cytosol of fungi and oomycetes. Here we present an in silico analysis of the tryptophan biosynthetic pathway in stramenopiles,\\u000a based on analysis of the genomes of the oomycetes Phytophthora sojae and P. ramorum and the diatoms Thalassiosira pseudonana and Phaeodactylum

Kate?ina Jiroutová; Aleš Horák; Chris Bowler; Miroslav Oborník



Survey and analysis of microsatellites from transcript sequences in Phytophthora species: frequency, distribution, and potential as markers for the genus  

Microsoft Academic Search

Background: Members of the genus Phytophthora are notorious pathogens\\u000a with world-wide distribution. The most devastating species include P.\\u000a infestans, P. ramorum and P. sojae. In order to develop molecular\\u000a methods for routinely characterizing their populations and to gain a\\u000a better insight into the organization and evolution of their genomes, we\\u000a used an in silico approach to survey and compare simple

Diana P. Garnica; Andres M. Pinzon; Lina M. Quesada-Ocampo; Adriana J. Bernal; Emiliano Barreto; Niklaus J. Grunwald; Silvia Restrepo



A New Cell Wall Located N-rich Protein is Strongly Induced During the Hypersensitive Response in Glycine Max L  

Microsoft Academic Search

Soybean (Glycine max (L.) Merill, cv. Williams 82) plants and cell cultures respond to avirulent pathogens with a hypersensitive reaction. After inoculation of soybean with Pseudomonas syringae pv. glycinea, carrying the avirulence gene avrA, or zoospores from the fungus Phytophthora sojae Race 1, a resistance-gene-dependent cell death programme is activated. A new gene was identified by differential display of mRNAs

Andrea A. Ludwig; Raimund Tenhaken



Map order and linkage distances of molecular markers close to the supernodulation ( nts-1 ) locus of soybean  

Microsoft Academic Search

The molecular characteristics of markers in the chromosome region surrounding the supernodulation gene (nts-1) of soybean (Glycine max L. Merr.) were investigated in 187 F2 plants from a cross of G. max cv. Bragg (nts) and G. soja PI468.397 (wild-type nodulation). RFLP marker pUTG-132a, linked tightly (0.7±0.5 cM) to nts-1, was converted to a PCR marker. The polymorphism resides within

A. Kolchinsky; D. Landau-Ellis; P. M. Gresshoff



Resistance of Glycine species and various cultivated legumes to the soybean aphid (Homoptera: Aphididae).  


The soybean aphid, Aphis glycines Matsumura, is a new pest of soybean, Glycine max (L.) Merr., in North America. It has become widespread on soybean in North America since it was first identified in the Midwest in 2000. Species of Rhamnus L. (buckthorn) are the primary hosts of A. glycines, and soybean is known as a secondary host. There is limited information about the secondary host range of A. glycines. Aphid colonization on various legume hosts was compared in choice experiments. Aphid colonization occurred on species in the genus Glycine Wild. No colonization occurred on Lablab purpureus (L.) Sweet, Lens culinaris Medik, Phaseolus vulgaris L., Pisum sativum L., or species of Vicia L. and Vigna Savi. Colonization was limited or aphids were transient on species of Medicago L., Phaseolus L., and Trifolium L. There were significant differences in aphid colonization among Medicago truncatula accessions with numbers ranging from 7 to 97 aphids per plant. Six Glycine soja Sieb. & Zucc. accessions were as resistant as G. max accessions to A. glycines; these may represent novel sources of A. glycines resistance not found in G. max. Antibiosis was found to play a large role in the expression of resistance in three of the G. soja accessions. Results of this study indicated that G. max and G. soja were the best secondary hosts of A. glycines; however, its secondary host range may include other leguminous species. Therefore, A. glycines did not seem to have a highly restricted monophagous secondary host range. PMID:15279293

Hill, Curtis B; Li, Yan; Hartman, Glen L



Distinct changes in soybean xylem sap proteome in response to pathogenic and symbiotic microbe interactions  

PubMed Central

Background Plant systemic signaling characterized by the long distance transport of molecules across plant organs involves the xylem and phloem conduits. Root-microbe interactions generate systemic signals that are transported to aerial organs via the xylem sap. We analyzed the xylem sap proteome of soybean seedlings in response to pathogenic and symbiotic interactions to identify systemic signaling proteins and other differentially expressed proteins. Results We observed the increase of a serine protease and peroxidase in the xylem sap in response to Phytophthora sojae elicitor treatment. The high molecular weight fraction of soybean xylem sap was found to promote the growth of Neurospora crassa in vitro at lower concentrations and inhibit growth at higher concentrations. Sap from soybean plants treated with a P. sojae elicitor had a significantly higher inhibitory effect than sap from control soybean plants. When soybean seedlings were inoculated with the symbiont Bradyrhizobium japonicum, the abundance of a xyloglucan transendoglycosyl transferase protein increased in the xylem sap. However, RNAi-mediated silencing of the corresponding gene did not significantly affect nodulation in soybean hairy root composite plants. Conclusion Our study identified a number of sap proteins from soybean that are differentially induced in response to B. japonicum and P. sojae elicitor treatments and a majority of them were secreted proteins.

Subramanian, Senthil; Cho, Un-Haing; Keyes, Carol; Yu, Oliver



Growth in microgravity increases susceptibility of soybean to a fungal pathogen.  


The influence of microgravity on the susceptibility of soybean roots to Phytophthora sojae was studied during the Space Shuttle Mission STS-87. Seedlings of soybean cultivar Williams 82 grown in spaceflight or at unit gravity were untreated or inoculated with the soybean root rot pathogen P. sojae. At 3, 6 and 7 d after launch while still in microgravity, seedlings were photographed and then fixed for subsequent microscopic analysis. Post-landing analysis of the seedlings revealed that at harvest day 7 the length of untreated roots did not differ between flight and ground samples. However, the flight-grown roots infected with P. sojae showed more disease symptoms (percentage of brown and macerated areas) and the root tissues were more extensively colonized relative to the ground controls exposed to the fungus. Ethylene levels were higher in spaceflight when compared to ground samples. These data suggest that soybean seedlings grown in microgravity are more susceptible to colonization by a fungal pathogen relative to ground controls. PMID:11427686

Ryba-White, M; Nedukha, O; Hilaire, E; Guikema, J A; Kordyum, E; Leach, J E



Intracellular and extracellular phosphatidylinositol 3-phosphate produced by phytophthora species is important for infection.  


RxLR effectors produced by Phytophthora pathogens have been proposed to bind to phosphatidylinositol 3-phosphate (PtdIns(3)P) to mediate their translocation into host cells and/or to increase their stability in planta. Since the levels of PtdIns(3)P in plants are low, we examined whether Phytophthora species may produce PtdIns(3)P to promote infection. We observed that PtdIns(3)P-specific GFP biosensors could bind to P. parasitica and P. sojae hyphae during infection of Nicotiana benthamiana leaves transiently secreting the biosensors, suggesting that the hyphae exposed PtdIns(3)P on their plasma membrane and/or secreted PtdIns(3)P. Silencing of the phosphatidylinositol 3-kinases (PI3K) genes, treatment with LY294002, or expression of PtdIns(3)P-binding proteins by P. sojae reduced the virulence of the pathogen on soybean, indicating that pathogen-synthesized PtdIns(3)P was required for full virulence. Secretion of PtdIns(3)P-binding proteins or of a PI3P-5-kinase by N. benthamiana leaves significantly increased the level of resistance to infection by P. parasitica and P. capsici. Together, our results support the hypothesis that Phytophthora species produce external PtdIns(3)P to aid in infection, such as to promote entry of RxLR effectors into host cells. Our results derived from P. sojae RxLR effector Avr1b confirm that both the N-terminus and the C-terminus of this effector can bind PtdIns(3)P. PMID:23475996

Lu, Shan; Chen, Linlin; Tao, Kai; Sun, Nannan; Wu, Yuren; Lu, Xiaoxue; Wang, Yuanchao; Dou, Daolong



The effects of microgravity and clinorotation on the interaction of plant cells with fungal pathogen  

NASA Astrophysics Data System (ADS)

The influence of microgravity and slow horizontal clinorotation (2 rev/min), which partly mimics microgravity, on the interaction of plant cells of soybean roots to Phytophthora sojae and of potato minitubers to Phytophthora infestans was studied during the Space Shuttle Mission STS-87 and during clinorotation. Seedlings of soybean cultivar Williams 82 grown in spaceflight and at 1 g were untreated or inoculated with pathogen P. sojae; minitubers of potato (cv Adreta) grown at horizontal clinorotation and the vertical control also were untreated or inoculated with pathogen P. infestans. The methods of light microscopy, scanning and transmission electron microscopy, confocal microscopy and also cytochemistry for the determination of callose content and peroxydase activity were used in the experiments. Post-landing analysis of the meristem cells of soybean roots infected with P. sojae and post-clinorotation analysis of the parenchyma cells of potato minitubers cells infected with P. infestans showed more destroying symptoms in cells of plant-host, which were more extensive colonized relative to the controls exposed to the pathogen fungus. Infected cells of plants-host were divided in two types: cells of first type were completely destroyed and hyphae of pathogen fungus were into these cells or in intercellular spaces; cells of second type characterized by partly changed ultrastructure and a calcium sites were contained above in mentioned cells. These data suggest that root cells of soybean seedlings grown in microgravity and cells of potato minitubers grown at slow horizontal clinorotation are more susceptible to penetration of a fungal pathogen in comparison with the corresponding controls.

Nedukha, O.; Kordyum, E.; Leach, J.; Martyn, G.; Ryba-White, M.


Phytophthora genome sequences uncover evolutionary origins and mechanisms of pathogenesis  

Microsoft Academic Search

Draft genome sequences have been determined for the soybean pathogen\\u000a Phytophthora sojae and the sudden oak death pathogen Phytophthora\\u000a ramorum. Oomycetes such as these Phytophthora species share the kingdom\\u000a Stramenopila with photosynthetic algae such as diatoms, and the presence\\u000a of many Phytophthora genes of probable phototroph origin supports a\\u000a photosynthetic ancestry for the stramenopiles. Comparison of the two\\u000a species' genomes

Brett M. Tyler; Sucheta Tripathy; Xuemin Zhang; Paramvir Dehal; Rays H. Y. Jiang; Andrea Aerts; Felipe D. Arredondo; Laura Baxter; Douda Bensasson; Jim L. Beynon; Jarrod Chapman; Cynthia M. B. Damasceno; Anne E. Dorrance; Daolong Dou; Allan W. Dickerman; Inna L. Dubchak; Matteo Garbelotto; Mark Gijzen; Stuart G. Gordon; Francine Govers; Niklaus J. Grunwald; Wayne Huang; Kelly L. Ivors; Richard W. Jones; Sophien Kamoun; Konstantinos Krampis; Kurt H. Lamour; Mi-Kyung Lee; W. Hayes McDonald; Mónica Medina; Harold J. G. Meijer; Eric K. Nordberg; Donald J. Maclean; Manuel D. Ospina-Giraldo; Paul F. Morris; Vipaporn Phuntumart; Nicholas H. Putnam; Sam Rash; Jocelyn K. C. Rose; Yasuko Sakihama; Asaf A. Salamov; Alon Savidor; Chantel F. Scheuring; Brian M. Smith; Bruno W. S. Sobral; Astrid Terry; Trudy A. Torto-Alalibo; Joe Win; Zhanyou Xu; Hongbin Zhang; Igor V. Grigoriev; Daniel S. Rokhsar; Jeffrey L. Boore



Effect of raw legume diets on intestinal absorption of D-galactose by chick.  


The effect of four raw legume diets on the intestinal absorption of D-galactose and oxygen consumption were studied in chick. Field beans (Vicia faba), soybeans (Glycine soja), bitter vetch (Vicia ervilia), and navy beans (Phaseolus vulgaris), were used. The intestinal absorption was determined by both in vivo and in vitro techniques. In vivo, only navy beans and soybeans inhibit intestinal transport of D-galactose, while in vitro all the diets do. Oxygen consumption by intestinal rings increases in chicks fed on bitter vetch diet. PMID:7394275

Lasheras, B; Bolufer, J; Cenarruzabeitia, M N; Lluch, M; Larralde, J



Intestinal absorption of D-galactose and L-leucine and intestinal disaccharidase activities in growing chickens fed different raw legume diets.  


A significant (P less than .01) impairment in the rate of growth, along with a significant (P less than .01) inhibition in the rate of in vivo intestinal absorption of D-galactose and L-leucine, and in the in vitro intestinal absorption of D-galactose, was found in growing chickens fed ad libitum over a 60-day period, diets containing the raw legumes Vicia faba, Glycine soja, Vicia ervilia, and Phaseolus vulgaris as the main source of protein. Furthermore, a significant (P less than .01) reduction in the intestinal disaccharidase activity was found in the legume-fed chickens. The possible nature of these effects was discussed. PMID:7301750

Santidrian, S; Lasheras, B; Cenarruzabeitia, M N; Bolufer, J; Larralde, J



Arabidopsis nonhost resistance gene PSS1 confers immunity against an oomycete and a fungal pathogen but not a bacterial pathogen that cause diseases in soybean  

PubMed Central

Background Nonhost resistance (NHR) provides immunity to all members of a plant species against all isolates of a microorganism that is pathogenic to other plant species. Three Arabidopsis thaliana PEN (penetration deficient) genes, PEN1, 2 and 3 have been shown to provide NHR against the barley pathogen Blumeria graminis f. sp. hordei at the prehaustorial level. Arabidopsis pen1-1 mutant lacking the PEN1 gene is penetrated by the hemibiotrophic oomycete pathogen Phytophthora sojae, the causal organism of the root and stem rot disease in soybean. We investigated if there is any novel nonhost resistance mechanism in Arabidopsis against the soybean pathogen, P. sojae. Results The P.sojaesusceptible (pss) 1 mutant was identified by screening a mutant population created in the Arabidopsis pen1-1 mutant that lacks penetration resistance against the non adapted barley biotrophic fungal pathogen, Blumeria graminis f. sp. hordei. Segregation data suggested that PEN1 is not epistatic to PSS1. Responses of pss1 and pen1-1 to P. sojae invasion were distinct and suggest that PSS1 may act at both pre- and post-haustorial levels, while PEN1 acts at the pre-haustorial level against this soybean pathogen. Therefore, PSS1 encodes a new form of nonhost resistance. The pss1 mutant is also infected by the necrotrophic fungal pathogen, Fusarium virguliforme, which causes sudden death syndrome in soybean. Thus, a common NHR mechanism is operative in Arabidopsis against both hemibiotrophic oomycetes and necrotrophic fungal pathogens that are pathogenic to soybean. However, PSS1 does not play any role in immunity against the bacterial pathogen, Pseudomonas syringae pv. glycinea, that causes bacterial blight in soybean. We mapped PSS1 to a region very close to the southern telomere of chromosome 3 that carries no known disease resistance genes. Conclusions The study revealed that Arabidopsis PSS1 is a novel nonhost resistance gene that confers a new form of nonhost resistance against both a hemibiotrophic oomycete pathogen, P. sojae and a necrotrophic fungal pathogen, F. virguliforme that cause diseases in soybean. However, this gene does not play any role in the immunity of Arabidopsis to the bacterial pathogen, P. syringae pv. glycinea, which causes bacterial blight in soybean. Identification and further characterization of the PSS1 gene would provide further insights into a new form of nonhost resistance in Arabidopsis, which could be utilized in improving resistance of soybean to two serious pathogens.



The Phytophthora Genome Initiative Database: informatics and analysis for distributed pathogenomic research  

PubMed Central

The Phytophthora Genome Initiative (PGI) is a distributed collaboration to study the genome and evolution of a particularly destructive group of plant pathogenic oomycete, with the goal of understanding the mechanisms of infection and resistance. NCGR provides informatics support for the collaboration as well as a centralized data repository. In the pilot phase of the project, several investigators prepared Phytophthora infestans and Phytophthora sojae EST and Phytophthora sojae BAC libraries and sent them to another laboratory for sequencing. Data from sequencing reactions were transferred to NCGR for analysis and curation. An analysis pipeline transforms raw data by performing simple analyses (i.e., vector removal and similarity searching) that are stored and can be retrieved by investigators using a web browser. Here we describe the database and access tools, provide an overview of the data therein and outline future plans. This resource has provided a unique opportunity for the distributed, collaborative study of a genus from which relatively little sequence data are available. Results may lead to insight into how better to control these pathogens. The homepage of PGI can be accessed at , with database access through the database access hyperlink.

Waugh, Mark; Hraber, Peter; Weller, Jennifer; Wu, Yihe; Chen, Guonghong; Inman, Jeff; Kiphart, Don; Sobral, Bruno



The phytophthora genome initiative database: informatics and analysis for distributed pathogenomic research.  


The Phytophthora Genome Initiative (PGI) is a distributed collaboration to study the genome and evolution of a particularly destructive group of plant pathogenic oomycete, with the goal of understanding the mechanisms of infection and resistance. NCGR provides informatics support for the collaboration as well as a centralized data repository. In the pilot phase of the project, several investigators prepared Phytophthora infestans and Phytophthora sojae EST and Phytophthora sojae BAC libraries and sent them to another laboratory for sequencing. Data from sequencing reactions were transferred to NCGR for analysis and curation. An analysis pipeline transforms raw data by performing simple analyses (i.e., vector removal and similarity searching) that are stored and can be retrieved by investigators using a web browser. Here we describe the database and access tools, provide an overview of the data therein and outline future plans. This resource has provided a unique opportunity for the distributed, collaborative study of a genus from which relatively little sequence data are available. Results may lead to insight into how better to control these pathogens. The homepage of PGI can be accessed at, with database access through the database access hyperlink. PMID:10592189

Waugh, M; Hraber, P; Weller, J; Wu, Y; Chen, G; Inman, J; Kiphart, D; Sobral, B



The aspartic proteinase family of three Phytophthora species  

PubMed Central

Background Phytophthora species are oomycete plant pathogens with such major social and economic impact that genome sequences have been determined for Phytophthora infestans, P. sojae and P. ramorum. Pepsin-like aspartic proteinases (APs) are produced in a wide variety of species (from bacteria to humans) and contain conserved motifs and landmark residues. APs fulfil critical roles in infectious organisms and their host cells. Annotation of Phytophthora APs would provide invaluable information for studies into their roles in the physiology of Phytophthora species and interactions with their hosts. Results Genomes of Phytophthora infestans, P. sojae and P. ramorum contain 11-12 genes encoding APs. Nine of the original gene models in the P. infestans database and several in P. sojae and P. ramorum (three and four, respectively) were erroneous. Gene models were corrected on the basis of EST data, consistent positioning of introns between orthologues and conservation of hallmark motifs. Phylogenetic analysis resolved the Phytophthora APs into 5 clades. Of the 12 sub-families, several contained an unconventional architecture, as they either lacked a signal peptide or a propart region. Remarkably, almost all APs are predicted to be membrane-bound. Conclusions One of the twelve Phytophthora APs is an unprecedented fusion protein with a putative G-protein coupled receptor as the C-terminal partner. The others appear to be related to well-documented enzymes from other species, including a vacuolar enzyme that is encoded in every fungal genome sequenced to date. Unexpectedly, however, the oomycetes were found to have both active and probably-inactive forms of an AP similar to vertebrate BACE, the enzyme responsible for initiating the processing cascade that generates the A? peptide central to Alzheimer's Disease. The oomycetes also encode enzymes similar to plasmepsin V, a membrane-bound AP that cleaves effector proteins of the malaria parasite Plasmodium falciparum during their translocation into the host red blood cell. Since the translocation of Phytophthora effector proteins is currently a topic of intense research activity, the identification in Phytophthora of potential functional homologues of plasmepsin V would appear worthy of investigation. Indeed, elucidation of the physiological roles of the APs identified here offers areas for future study. The significant revision of gene models and detailed annotation presented here should significantly facilitate experimental design.



Studies on lectins. XL. O-glycosyl derivatives of Spheron in affinity chromatography of lectins.  


Free monosaccharides can be used for direct glycosylation of Spheron, a spherical macroporous hydroxyalkyl methacrylate-ethylene dimethacrylate copolymer, in a reaction that proceeds at room temperature in dioxane medium under catalysis of dry HCl or BF3. Derivatives of L-fucose, D-galactose, D-glucose, D-mannose, N-acetyl-D-galactosamine and N-acetyl-D-glucosamine thus prepared from Spheron beads have been shown to be efficient affinity carriers in isolation of lectins from seeds of Canavalia ensiformis D.C. (concanavalin A), Dolichos biflorus L., Glycine soja (L.) Sieb. et Zucc., Lens esculenta Moench, Ricinus communis L., Ulex europaeus L. and from albumin glands of the garden snail Helix pomatia L. PMID:638203

Filka, K; Coupek, J; Kocourek, J



Effect of the raw legume Vicia ervilia on muscle and liver protein metabolism in growing rats.  


Body weight gain (BWG), food intake, food efficiency rate (FER: food intake, g/protein intake, g), gastrocnemius muscle and liver weights, protein, RNA and DNA contents of gastrocnemius muscle and liver have been measured in growing rats (80-90 g initial body weight) fed ad libitum over a period of 11 days on 12.00% protein diets containing either heated and defatted Glycinae sojae (HSB) as control or the raw legume Vicia ervilia as the main sources of protein. It has been found that, as compared to HSB-fed rats, those fed the legume Vicia ervilia diet exhibited a significant reduction in growth, PER, FER, as well as, in RNA-activity (protein, g/day/RNA, g) and RNA/DNA ratio in both muscle and liver. Protein synthesis capacity (PSC:RNA, microgram/protein, mg), was found significantly increased in liver but not in muscle. The possible nature of these findings is discussed. PMID:2484317

Goena, M; Marzo, F; Fernández-González, A L; Tosar, A; Frühbeck, G; Santidrián, S



Effect of raw legume diets on disaccharidase activity in the small intestine of chicks.  


The effect of four raw legume diets: field beans (Vicia faba) (RFB), navy beans (Phaseolus vulgaris) (RNB), soybeans (Glycine soja) (RSB) and bitter vetch (VICIA ervilia) (RBV), on disaccharidase activities in chick small intestine have been studied. Maltase and sucrase activities, which vary with age, were determined in 1 to 60 day old animals, RFB and RBV diets had no effect on maltase activity and only increased sucrase activity in 60 day old chicks. Both maltase and sucrase activities decreased in chicks on RSB diet, regardless of their age, and the decrease was even more pronounced in chicks on RNB diet. Contrarywise, chicks fed on autoclaved navy beans and soybeans showed a considerably higher activity of these disaccharidases. PMID:7192009

Lasheras, B; Cenarruzabeitia, M N; Fontán, J; Lluch, M; Larralde, J



Structure and chromosomal arrangement of leghemoglobin genes in kidney bean suggest divergence in soybean leghemoglobin gene loci following tetraploidization  

PubMed Central

We have determined the structure of one of the leghemoglobin (Lb) genes of Phaseolus vulgaris (kidney bean) and deduced the chromosomal arrangement of leghemoglobin genes by genomic hybridizations with Lb and two other sequences, each specific to the 5' or 3' region of the soybean leghemoglobin loci. By comparing this organization with two other species of legumes, Glycine max (soybean) and G. soja (wild soybean), a phylogeny of leghemoglobin gene loci was traced. The intragenic structure of the kidney bean leghemoglobin gene shows the same intron/exon arrangement as that of soybean leghemoglobin genes and extensive sequence homologies in both coding as well as 5' and 3' non-coding regions. The presence in the kidney bean genome of four leghemoglobin genes suggests that tandem duplications of a single primordial plant globin gene had occurred to generate four leghemoglobin genes in an `Lb-locus' before Glycine and Phaseolus species diverged. Chromosome duplication by tetraploidization in Glycine generated two loci containing four genes each. A large deletion in one of the two four-gene loci in soybean resulted in the generation of the Lbc2 locus containing two leghemoglobin genes, one functional and another pseudo (Lb?2). This pseudogene, unlike that present on the main locus, is represented by only two and a half exons and appears to be truncated. The two other truncated genes (LbT1 and LbT2) were probably generated similarly in the genome of Glycine spp. following tetraploidization before the divergence of G. max and G. soja. ImagesFig. 4.Fig. 5.Fig. 6.

Lee, Jong Seob; Verma, Desh Pal S.



Analyses of genome architecture and gene expression reveal novel candidate virulence factors in the secretome of Phytophthora infestans  

PubMed Central

Background Phytophthora infestans is the most devastating pathogen of potato and a model organism for the oomycetes. It exhibits high evolutionary potential and rapidly adapts to host plants. The P. infestans genome experienced a repeat-driven expansion relative to the genomes of Phytophthora sojae and Phytophthora ramorum and shows a discontinuous distribution of gene density. Effector genes, such as members of the RXLR and Crinkler (CRN) families, localize to expanded, repeat-rich and gene-sparse regions of the genome. This distinct genomic environment is thought to contribute to genome plasticity and host adaptation. Results We used in silico approaches to predict and describe the repertoire of P. infestans secreted proteins (the secretome). We defined the "plastic secretome" as a subset of the genome that (i) encodes predicted secreted proteins, (ii) is excluded from genome segments orthologous to the P. sojae and P. ramorum genomes and (iii) is encoded by genes residing in gene sparse regions of P. infestans genome. Although including only ~3% of P. infestans genes, the plastic secretome contains ~62% of known effector genes and shows >2 fold enrichment in genes induced in planta. We highlight 19 plastic secretome genes induced in planta but distinct from previously described effectors. This list includes a trypsin-like serine protease, secreted oxidoreductases, small cysteine-rich proteins and repeat containing proteins that we propose to be novel candidate virulence factors. Conclusions This work revealed a remarkably diverse plastic secretome. It illustrates the value of combining genome architecture with comparative genomics to identify novel candidate virulence factors from pathogen genomes.



Isolation and characterization of rhizobitoxine mutants of Bradyrhizobium japonicum.  

PubMed Central

To explore the role of rhizobitoxine in Bradyrhizobium-legume symbiosis, 11 rhizobitoxine mutants of B. japonicum USDA61 were isolated on the basis of their inability to synthesize the toxin in culture. Each mutant is prototrophic and symbiotically effective on soybean, cowpea, siratro, and Glycine soja. The rhizobitoxine mutants differ in their chlorosis phenotypes and rhizobitoxine production in planta. As expected, one group of mutant fail to make toxin in planta, resulting in the absence of chlorosis. Another group of mutants causes severe chlorosis on all cultivars of soybean tested. Surprisingly, this group of mutants makes more rhizobitoxine in soybean nodules than the wild-type strain does. This phenotype is only observed on soybean and not on other hosts such as cowpea, siratro, or G. soja. The remaining mutants all produce rhizobitoxine in planta but vary in the amount of toxin they produce and the severity of chlorosis they induce in soybean plants. Biochemical analysis of mutants demonstrates that one mutant is unable to synthesize serinol, a molecule hypothesized to be an intermediate in rhizobitoxine biosynthesis. By using these mutants, it was found that rhizobitoxine plays no apparent role in the nodulation of rj1 soybeans. Recently, it was found that inhibition of ethylene biosynthesis allows Rhizobium meliloti to overcome nitrate inhibition of nodule formation on alfalfa. Because rhizobitoxine also inhibits ethylene biosynthesis, we tested the ability of mutants which accumulate high levels of toxin in planta to overcome nitrate inhibition of nodule formation on soybean plants and found that the nodule formation induced by the wild type and that induced by mutant strains were equally suppressed in the presence of nitrate. Images

Ruan, X; Peters, N K



Identification and candidate gene analysis of a novel phytophthora resistance gene Rps10 in a Chinese soybean cultivar.  


Resistance to Phytophthora sojae isolate PsMC1 was evaluated in 102 F2?3 families derived from a cross between the resistant soybean cultivar Wandou 15 and the susceptible cultivar Williams and genotyped using simple sequence repeat (SSR) markers. The segregation ratio of resistant, segregating, and susceptible phenotypes in the population suggested that the resistance in Wandou 15 was dominant and monogenic. Twenty-six polymorphic SSR markers were identified on soybean chromosome 17 (Molecular linkage group D2; MLG D2), which were linked to the resistance gene based on bulked segregation analysis (BSA). Markers Sattwd15-24/25 and Sattwd15-47 flanked the resistance gene at a distance of 0.5 cM and 0.8 cM, respectively. Two cosegregating markers, Sattwd15-28 and Sattwd15-32, were also screened in this region. This is the first Rps resistance gene mapped on chromosome 17, which is designated as Rps10. Eight putative genes were found in the mapped region between markers Sattwd15-24/25 and Sattwd15-47. Among them, two candidate genes encoding serine/threonine (Ser/Thr) protein kinases in Wandou 15 and Williams were identified and sequenced. And the differences in genomic sequence and the putative amino acid sequence, respectively, were identified within each candidate gene between Wandou 15 and Williams. This novel gene Rps10 and the linked markers should be useful in developing soybean cultivars with durable resistance to P. sojae. PMID:23936102

Zhang, Jiqing; Xia, Changjian; Duan, Canxing; Sun, Suli; Wang, Xiaoming; Wu, Xiaofei; Zhu, Zhendong



Chronostratigraphical Subdivision of the Late Glacial and the Holocene for the Alaska Region  

NASA Astrophysics Data System (ADS)

Our work is a kind of so called data mining. The first step of our work was collection of the radiocarbon data for samples coming from Alaska. We construct data base using Radiocarbon Measurements Lists published by different radiocarbon laboratories (mainly in the journal 'Radiocaron'). The next step was careful analysis of collected dates. We excluded from our analysis all dates suspected of contamination by younger or older organic matter. Such fact could be stated, for instance, on the base of inconsistency of radiocarbon age and stratigraphy or palynology. Finally, we calibrated whole large set of chosen radiocarbon dates and construct probability density function (PDF). Analysis of the shape of PDF was the subject of the previous research (eg. Michczynska and Pazdur, 2004; Macklin et al., 2006; Starkel et al., 2006, Michczynska et al., 2007). In our analysis we take into account the distinct tendency to collect samples from specific horizons. It is a general rule to take samples for radiocarbon dating from places of visible sedimentation changes or changes in palynological diagram. Therefore the culminations of the PDF represent periods of environmental changes and could be helpful in identifying the chronostratigraphical boundaries on the calendar time scale. References: Michczy?ska D.J., Pazdur A., 2004. A shape analysis of cumulative probability density function of radiocarbon dates set in the study of climate change in Late Glacial and Holocene. Radiocarbon 46(2): 733-744. Michczy?ska D.J., Michczy?ski A., Pazdur A. 2007. Frequency distribution of radiocarbon dates as a tool for reconstructing environmental changes. Radiocarbon 49(2): 799-806. Macklin M.G., Benito G., Gregory K.J., Johnstone E., Lewin J., Michczy?ska D.J., Soja R., Starkel L., Thorndycraft V.R., 2006. Past hydrological events reflected in the Holocene fluvial record of Europe. CATENA 66: 145-154. Starkel L., Soja R., Michczy?ska D.J., 2006. Past hydrological events reflected in Holocene history of Polish rivers. CATENA 66: 24-33.

Michczynska, D. J.; Hajdas, I.



Profiling of dynamic changes in the microbial community during the soy sauce fermentation process.  


Soy sauce is a traditional condiment manufactured by natural inoculation and mixed culture fermentation. As is well known, it is the microbial community that plays an important role in the formation of its flavors. However, to date, its dynamic changes during the long period of fermentation process are still unclear, intensively constraining the improvement and control of the soy sauce quality. In this work, we revealed the dynamic changes of the microbial community by combining a cultured dependent method and a cultured independent method of polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis. Results indicated that the two methods verified and complemented each other in profiling microbial community, and that significant dynamics of the microbial community existed during the fermentation process, especially the strong inhibition of the growth of most of the microbes when entering into the mash stage from the koji stage. In the analysis of bacterial community, Staphylococcus and Bacillus were found to be the dominant bacteria and detected in the whole fermentation process. Kurthia and Klebsiella began to appear in the koji stage and then fade away in the early stage of the mash fermentation. In the analysis of fungal community, Aspergillus sojae and Zygosaccharomyces rouxii were found to be the dominant fungi in the koji and mash fermentation, respectively. It was clearly shown that when A. sojae decreased and disappeared in the middle stage of the mash fermentation, Z. rouxii appeared and increased at the meantime. Aspergillus parasiticus, Trichosporon ovoides and Trichosporon asahii also appeared in the koji and the early period of the mash fermentation and disappeared thereafter. Similar to Z. rouxii, Millerozyma farinosa and Peronospora farinosa were also found spontaneously which appeared in the mid-late period of the mash fermentation. The principal component analysis suggested that the microbial community underwent significant changes in the early period of the fermentation and, thereafter, tended to the stabilization in the mid-late periods. This study gave us important clues to understand the fermentation process and can serve as a foundation for improving the quality of soy sauce in the future. PMID:24037306

Wei, Quanzeng; Wang, Hongbin; Chen, Zhixin; Lv, Zhijia; Xie, Yufeng; Lu, Fuping



The soybean-Phytophthora resistance locus Rps1-k encompasses coiled coil-nucleotide binding-leucine rich repeat-like genes and repetitive sequences  

PubMed Central

Background A series of Rps (resistance to Pytophthora sojae) genes have been protecting soybean from the root and stem rot disease caused by the Oomycete pathogen, Phytophthora sojae. Five Rps genes were mapped to the Rps1 locus located near the 28 cM map position on molecular linkage group N of the composite genetic soybean map. Among these five genes, Rps1-k was introgressed from the cultivar, Kingwa. Rps1-k has been providing stable and broad-spectrum Phytophthora resistance in the major soybean-producing regions of the United States. Rps1-k has been mapped and isolated. More than one functional Rps1-k gene was identified from the Rps1-k locus. The clustering feature at the Rps1-k locus might have facilitated the expansion of Rps1-k gene numbers and the generation of new recognition specificities. The Rps1-k region was sequenced to understand the possible evolutionary steps that shaped the generation of Phytophthora resistance genes in soybean. Results Here the analyses of sequences of three overlapping BAC clones containing the 184,111 bp Rps1-k region are reported. A shotgun sequencing strategy was applied in sequencing the BAC contig. Sequence analysis predicted a few full-length genes including two Rps1-k genes, Rps1-k-1 and Rps1-k-2. Previously reported Rps1-k-3 from this genomic region [1] was evolved through intramolecular recombination between Rps1-k-1 and Rps1-k-2 in Escherichia coli. The majority of the predicted genes are truncated and therefore most likely they are nonfunctional. A member of a highly abundant retroelement, SIRE1, was identified from the Rps1-k region. The Rps1-k region is primarily composed of repetitive sequences. Sixteen simple repeat and 63 tandem repeat sequences were identified from the locus. Conclusion These data indicate that the Rps1 locus is located in a gene-poor region. The abundance of repetitive sequences in the Rps1-k region suggested that the location of this locus is in or near a heterochromatic region. Poor recombination frequencies combined with presence of two functional Rps genes at this locus has been providing stable Phytophthora resistance in soybean.

Gao, Hongyu; Bhattacharyya, Madan K



Signaling in Soybean Phenylpropanoid Responses (Dissection of Primary, Secondary, and Conditioning Effects of Light, Wounding, and Elicitor Treatments).  

PubMed Central

The spatial and temporal deployment of plant defense responses involves a complex interplay of signal events, often resulting in superimposition of signaling processes. We have employed a minimal-wound protocol to clearly separate and characterize the specific contributions of light, wounding, and a wall glucan elicitor preparation (PWG) from Phytophthora sojae (Kauf. and Gerde.) to the regulation of phenylpropanoid defense responses in soybean (Glycine max L. [Merr.]) cotyledon tissues. The assay also allowed us to clearly reconstitute responses to combinations of these primary signals and to examine the effects of other pathogenesis-related molecules on the responses in a defined manner. Light specifically triggers accumulation of malonylglucosyl conjugates of the 5-hydroxy-isoflavone, genistein, which is normally found in epidermal cells. PWG selectively induces accumulation of conjugates of the 5-deoxy-isoflavone daidzein, the first committed precursor of the phytoalexin glyceollin. Wounding initiates phenolic polymer deposition, a process greatly potentiated by PWG and light. Whereas glutathione selectively enhances light induction of genistein conjugates, methyl jasmonate enhances both light and PWG-induced isoflavone conjugate accumulations. Wound exudate fully activates the cell's capacity (competency) for the phenolic polymer and glyceollin responses to PWG, whereas glutathione partially restores competency, favoring coumestrol and phenolic polymer responses to PWG. Abscisic acid inhibits all induced phenylpropanoid responses.

Graham, T. L.; Graham, M. Y.



Effects of light treatment on isoflavone content of germinated soybean seeds.  


Our research objective was to increase isoflavone content in the germinated soybean seeds of four different varieties (Pungsannamulkong, Cheongjakong, Aga4, and Aga3) by optimizing light treatments (dark, greenhouse, fluorescent, incandescent, and ultraviolet lamps). The results demonstrated that the highest isoflavone content was produced from the Aga3 variety, which was developed by an interspecific cross between Eunhakong (Glycine max) and KLG10084 (G. soja) at the Plant Genetic Laboratory, Kyunpook National University. Aga3 is known to have one of the highest isoflavone content in the world at present. Our results recommend exposure of 7-day-old Aga3 sprouts to a combined light treatment of greenhouse lamps (12 h per day) and ultraviolet light (40 min per day) for maximum isoflavone production. Aga3 produced high levels of isoflavone because, in part, it contained very high isoflavone levels within the seed as compared with the other varieties. Under stress conditions, Aga3 could produce over 1.90 times more isoflavone than its seed content and 1.53 times more isoflavone than when grown under dark conditions. PMID:18841981

Phommalth, Siviengkhek; Jeong, Yeon-Shin; Kim, Yong-Hoon; Dhakal, Krishna Hari; Hwang, Young-Hyun



The ?-conglycinin deficiency in wild soybean is associated with the tail-to-tail inverted repeat of the ?-subunit genes.  


?-conglycinin, a major seed protein in soybean, is composed of ?, ?', and ? subunits sharing a high homology among them. Despite its many health benefits, ?-conglycinin has a lower amino acid score and lower functional gelling properties compared to glycinin, another major soybean seed protein. In addition, the ?, ?', and ? subunits also contain major allergens. A wild soybean (Glycine soja Sieb et Zucc.) line, 'QT2', lacks all of the ?-conglycinin subunits, and the deficiency is controlled by a single dominant gene, Scg-1 (Suppressor of ?-conglycinin). This gene was characterized using a soybean cultivar 'Fukuyutaka', 'QY7-25', (its near-isogenic line carrying the Scg-1 gene), and the F? population derived from them. The physical map of the Scg-1 region covered by lambda phage genomic clones revealed that the two ?-subunit genes, a ?-subunit gene, and a pseudo ?-subunit gene were closely organized. The two ?-subunit genes were arranged in a tail-to-tail orientation, and the genes were separated by 197 bp in Scg-1 compared to 3.3 kb in the normal allele (scg-1). In addition, small RNA was detected in immature seeds of the mutants by northern blot analysis using an RNA probe of the ? subunit. These results strongly suggest that ?-conglycinin deficiency in QT2 is controlled by post-transcriptional gene silencing through the inverted repeat of the ? subunits. PMID:22193750

Tsubokura, Yasutaka; Hajika, Makita; Kanamori, Hiroyuki; Xia, Zhengjun; Watanabe, Satoshi; Kaga, Akito; Katayose, Yuichi; Ishimoto, Masao; Harada, Kyuya



Lack of galactose or galacturonic acid in Bradyrhizobium japonicum USDA 110 exopolysaccharide leads to different symbiotic responses in soybean.  


Exopolysaccharide (EPS) and lipopolysaccharide (LPS) from Bradyrhizobium japonicum are important for infection and nodulation of soybean (Glycine max), although their roles are not completely understood. To better understand this, we constructed mutants in B. japonicum USDA 110 impaired in galactose or galacturonic acid incorporation into the EPS without affecting the LPS. The derivative LP 3010 had a deletion of lspL-ugdH and produced EPS without galacturonic acid whereas LP 3013, with an insertion in exoB, produced EPS without galactose. In addition, the strain LP 3017, with both mutations, had EPS devoid of both galactosides. The missing galactosides were not replaced by other sugars. The defects in EPS had different consequences. LP 3010 formed biofilms and nodulated but was defective in competitiveness for nodulation; and, inside nodules, the peribacteroid membranes tended to fuse, leading to the merging of symbiosomes. Meanwhile, LP 3013 and LP 3017 were unable to form biofilms and produced empty pseudonodules but exoB suppressor mutants were obtained when LP 3013 plant inoculation was supplemented with wild-type EPS. Similar phenotypes were observed with all these mutants in G. soja. Therefore, the lack of each galactoside in the EPS has a different functional effect on the B. japonicum-soybean symbiosis. PMID:20636103

Quelas, Juan Ignacio; Mongiardini, Elías J; Casabuono, Adriana; López-García, Silvina L; Althabegoiti, M Julia; Covelli, Julieta M; Pérez-Giménez, Julieta; Couto, Alicia; Lodeiro, Aníbal R



Genome sequences of two Phytophthora species responsible for Sudden Oak Death and Soybean Root Rot provide novel insights into their evolutionary origins and mechanisms of pathogenesis  

SciTech Connect

The approximately 60 species of Phytophthora are all destructive pathogens, causing rots of roots, stems, leaves and fruits of a wide range of agriculturally and ornamentally important plants (1). Some species, such as P. cinnamomi, P. parasitica and P. cactorum, each attack hundreds of different plant host species, whereas others are more restricted. Some of the crops where Phytophthora infections cause the greatest financial losses include potato, soybean, tomato, alfalfa, tobacco, peppers, cucurbits, pineapple, strawberry, raspberry and a wide range of perennial tree crops, especially citrus, avocado, almonds, walnuts, apples and cocoa, and they also heavily affect the ornamental, nursery and forestry industries. The economic damage overall to crops in the United States by Phytophthora species is estimated in the tens of billions of dollars, including the costs of control measures, and worldwide it is many times this amount (1). In the northern midwest of the U.S., P. sojae causes $200 million in annual losses to soybean alone, and worldwide causes around $1-2 billion in losses per year. P. infestans infections resulted in the Irish potato famine last century and continues to be a difficult and worsening problem for potato and tomato growers worldwide, with worldwide costs estimated at $5 billion per year.

Tyler, Brett M.; Tripathi, Sucheta; Aerts, Andrea; Bensasson, Douda; Dehal, Paramvir; Dubchak, Inna; Garbelotto, Matteo; Gijzen, Mark; Huang, Wayne; Ivors, Kelly; Jiang, Rays; Kamoun, Sophien; Krampis, Konstantinos; Lamour, Kurt; McDonald, Hayes; Medina, Monica; Morris, Paul; Putnam, Nik; Rash, Sam; Salamov, Asaf; Smith, Brian; Smith, Joe; Terry, Astrid; Torto, Trudy; Grigoriev, Igor; Rokhsar, Daniel; Boore, Jeffrey



Host-Pathogen Interactions  

PubMed Central

The invertase present in the culture fluid of races 1, 2, and 3 of Phytophthora megasperma Drechs. var. sojae A. A. Hildebrand (Pms) were purified until they gave but a single band, whether stained for protein or carbohydrate, after isoelectric focusing in flat bed gels. The sugar compositions of multiple preparations of the purified invertases from each race of this fungal pathogen were determined by quantitative gas chromatography of their alditol acetates. The invertases are composed of about 25% carbohydrate. Mannose and glucosamine make up more than 97% of the carbohydrate portions of the invertases of all three Pms races analyzed, but the ratio of mannose to glucosamine is clearly not the same in each race. The glycosyl linkage compositions of the glucosamine-containing mannans of multiple preparations of the Pms invertases were determined by GC-MS analysis of the partially methylated alditol acetate derivatives. The results of these analyses demonstrate clear quantitative differences between the glycosyl components of the different Pms races. The existence of race-specific carbohydrate structures in the differentially virulent Pms races suggests that these carbohydrates may be involved in determining the specificity of hostpathogen interactions.

Ziegler, Ernst; Albersheim, Peter



Construction and characterization of a DNA probe for distinguishing strains of Aspergillus flavus.  

PubMed Central

Repetitive DNA sequences have proven useful and reliable characters in evaluating genetic relatedness of strains at different levels of taxonomic classification. A DNA probe was constructed to distinguish among strains of Aspergillus flavus by DNA fingerprinting techniques. Chromosomal DNA of A. flavus var. flavus NRRL 6541 was partially digested with EcoRI and ligated to a Lambda Dash bacteriophage vector. Four lambda clones were identified which displayed multiple and distinct bands when hybridized with chromosomal DNA from seven strains of A. flavus var. flavus digested with either EcoRI or PstI. One of these clones was chosen for further analysis and was subcloned into pUC19. The subclone, pAF28, contained a 6.2-kb chromosomal DNA insert and was able to distinguish among strains characterized by K. E. Papa (Mycologia 78:98-101, 1986) as belonging to 22 different vegetative compatibility groups. The subclone identified unique banding patterns when hybridized to genomic DNA digested with PstI. The cloned probe may be species specific as it hybridized with the DNA of all isolates of A. flavus tested in addition to strains recognized as varieties of A. flavus (e.g., A. flavus var. oryzae, A. flavus var. parasiticus, and A. flavus var. sojae). pAF28 hybridized to a single band on a Southern blot with Aspergillus nomius DNA but did not hybridize with the DNA of other fungal species tested including Aspergillus ochraceus, Aspergillus auricomus, Aspergillus alliaceus, Fusarium moniliforme, and Penicillium thomii.

McAlpin, C E; Mannarelli, B



Cloning, heterologous expression, and functional characterization of a chitinase gene, Lbchi32, from Limonium bicolor.  


In the present study, an endochitinase gene, Lbchi32, was cloned from Limonium bicolor. The cDNA sequence of Lbchi32 was 1,443 bp in length and encoded 319 amino acid residues. The DNA sequence of Lbchi32 was 2,512 bp in length and contained three exons and two introns. The Lbchi32 gene was inserted into a pPIC9 vector and transferred into Pichia pastoris strains GS115 and KM71 for heterologous expression. SDS-PAGE analyses indicated that LbCHI32 was expressed in both GS115 and KM71 and that it was secreted extracellularly. The optimal reaction conditions for LbCHI32 activity are 45 degrees C, pH 5.0, and 5 mM Ba(2+). The LbCHI32 enzyme can efficiently degrade chitin, chitin derivatives, and the cell walls of different pathogenic fungi, including phytopathogenic Rhizoctonia solani, Fusarium oxysporum, Sclerotinia sclerotiorum, Valsa sordida, Septoria tritici, and Phytophthora sojae. These findings suggest that Lbchi32 has potential use in the degradation of chitin and chitin derivatives. PMID:20512617

Liu, Zhi Hua; Yang, Chuan Ping; Qi, Xiao Tian; Xiu, Li Li; Wang, Yu Cheng



Cloning and characterization of a chitinase gene Lbchi31 from Limonium bicolor and identification of its biological activity.  


Chitinases are digestive enzymes that break down glycosidic bonds in chitin. In the current study, an endochitinase gene Lbchi31 was cloned from Limonium bicolor. The cDNA sequence of Lbchi31 was 1,107 bp in length, encoding 322 amino acid residues with a calculated molecular mass of 31.7 kDa. Clustal analysis showed that there was a highly conserved chitin-binding domains in Lbchi31 protein, containing four sulfide bridges. The Lbchi31 gene was inserted into the pPIC9 vector and transferred into yeast Pichia pastoris GS115 and KM71 for heterologous expression. The transformant harboring the Lbchi31 gene showed a clearly visible protein band with a molecular mass of more than 31 kDa in the SDS-PAGE gel, indicating that it had been translated in P. pastoris. Enzyme characterization showed that the optimal reaction condition for chitinase LbCHI31 activity was: 40 degrees C, pH of 5.0 and 5 mmol l(-1) of Mn(2+). The maximum enzyme activity was 0.88 U ml(-1) following exposure to the cell wall chitin of Valsa sordida. The LbCHI31 enzyme can efficiently degrade cell wall chitin of the phytopathogenic Rhizoctonia solani, Fusarium oxysporum, Sclerotinia sclerotiorum, V. sordida, Septoria tritici and Phytophthora sojae, suggesting that it has the biocontrol function to fungal phytopathogen. PMID:19685157

Liu, Zhi Hua; Wang, Yu Cheng; Qi, Xiao Tian; Yang, Chuan Ping



Competence for Elicitation of H2O2 in Hypocotyls of Cucumber Is Induced by Breaching the Cuticle and Is Enhanced by Salicylic Acid.  

PubMed Central

To study H2O2 production, the epidermal surfaces of hypocotyl segments from etiolated seedlings of cucumber (Cucumis sativus L.) were gently abraded. Freshly abraded segments were not constitutively competent for rapid H2O2 elicitation. This capacity developed subsequent to abrasion in a time-dependent process that was greatly enhanced in segments exhibiting an acquired resistance to penetration of their epidermal cell walls by Colletotrichum lagenarium, because of root pretreatment of the respective seedlings with 2,6-dichloroisonicotinic acid. When this compound or salicylic acid was applied to abraded segments, it also greatly enhanced the induction of competence for H2O2 elicitation. This process was fully inhibited by 5 [mu]M cycloheximide or 200 [mu]M puromycin, suggesting a requirement for translational protein synthesis. Both a crude elicitor preparation and a partially purified oligoglucan mixture from Phytophthora sojae also induced, in addition to H2O2 production, a refractory state, which explains the transient nature of H2O2 elicitation. Taken together, these results suggest that the cucumber hypocotyl epidermis becomes conditioned for competence to produce H2O2 in response to elicitors by a stimulus resulting from breaching the cuticle and/or cutting segments. This conditioning process is associated with protein synthesis and is greatly enhanced when substances able to induce systemic acquired resistance are present in the tissue.

Fauth, M.; Merten, A.; Hahn, M. G.; Jeblick, W.; Kauss, H.



[Antimicrobial efficacy of benzyl isothiocyanate].  


The potential value of benzyl isothiocyanate (BIT) vapor for food preservation was investigated in comparison with allyl isothiocyanate (AIT) vapor. Measurements of minimum inhibitory concentration (MIC) against various bacteria and fungi indicated that BIT vapor shows higher antibacterial activity towards Gram-positive bacteria, including lactic acid bacteria, than AIT vapor. BIT has a similar inhibitory effect to AIT against Gram-negative bacteria, yeast and fungi, except that it was less effective against Pseudomonas fluorescens JCM 5963. Both BIT and AIT have a strong inhibitory activity against yeast and fungi. AIT vapor suppressed growth of total aerobic bacteria in bigeye tuna meat and pork mince during storage at 10 degrees C and 30 degrees C, while BIT vapor had no effect. Also, AIT vapor strongly inhibited mycelial development of Aspergillus sojae JCM 2251 and Penicillium expansum IAM 13777 inoculated on rice cakes, while BIT vapor only slightly suppressed their growth. BIT has antimicrobial activities, but further studies are needed to establish its suitability for use in food preservation. PMID:20065621

Kamii, Eri; Isshiki, Kenji



Uncovering the Salt Response of Soybean by Unraveling Its Wild and Cultivated Functional Genomes Using Tag Sequencing  

PubMed Central

Soil salinity has very adverse effects on growth and yield of crop plants. Several salt tolerant wild accessions and cultivars are reported in soybean. Functional genomes of salt tolerant Glycine soja and a salt sensitive genotype of Glycine max were investigated to understand the mechanism of salt tolerance in soybean. For this purpose, four libraries were constructed for Tag sequencing on Illumina platform. We identify around 490 salt responsive genes which included a number of transcription factors, signaling proteins, translation factors and structural genes like transporters, multidrug resistance proteins, antiporters, chaperons, aquaporins etc. The gene expression levels and ratio of up/down-regulated genes was greater in tolerant plants. Translation related genes remained stable or showed slightly higher expression in tolerant plants under salinity stress. Further analyses of sequenced data and the annotations for gene ontology and pathways indicated that soybean adapts to salt stress through ABA biosynthesis and regulation of translation and signal transduction of structural genes. Manipulation of these pathways may mitigate the effect of salt stress thus enhancing salt tolerance.

Ali, Zulfiqar; Zhang, Da Yong; Xu, Zhao Long; Xu, Ling; Yi, Jin Xin; He, Xiao Lan; Huang, Yi Hong; Liu, Xiao Qing; Khan, Asif Ali; Trethowan, Richard M.; Ma, Hong Xiang



Receptor-Mediated Increase in Cytoplasmic Free Calcium Required for Activation of Pathogen Defense in Parsley  

PubMed Central

Transient influx of Ca2+ constitutes an early element of signaling cascades triggering pathogen defense responses in plant cells. Treatment with the Phytophthora sojae–derived oligopeptide elicitor, Pep-13, of parsley cells stably expressing apoaequorin revealed a rapid increase in cytoplasmic free calcium ([Ca2+]cyt), which peaked at ?1 ?M and subsequently declined to sustained values of 300 nM. Activation of this biphasic [Ca2+]cyt signature was achieved by elicitor concentrations sufficient to stimulate Ca2+ influx across the plasma membrane, oxidative burst, and phytoalexin production. Sustained concentrations of [Ca2+]cyt but not the rapidly induced [Ca2+]cyt transient peak are required for activation of defense-associated responses. Modulation by pharmacological effectors of Ca2+ influx across the plasma membrane or of Ca2+ release from internal stores suggests that the elicitor-induced sustained increase of [Ca2+]cyt predominantly results from the influx of extracellular Ca2+. Identical structural features of Pep-13 were found to be essential for receptor binding, increases in [Ca2+]cyt, and activation of defense-associated responses. Thus, a receptor-mediated increase in [Ca2+]cyt is causally involved in signaling the activation of pathogen defense in parsley.

Blume, Beatrix; Nurnberger, Thorsten; Nass, Norbert; Scheel, Dierk



GK4, a G-protein-coupled receptor with a phosphatidylinositol phosphate kinase domain in Phytophthora infestans, is involved in sporangia development and virulence.  


For dispersal and host infection plant pathogens largely depend on asexual spores. Pathogenesis and sporulation are complex processes that are governed by cellular signalling networks including G-protein and phospholipid signalling. Oomycetes possess a family of novel proteins called GPCR-PIPKs (GKs) that are composed of a seven-transmembrane spanning (7-TM) domain fused to a phosphatidylinositol phosphate kinase (PIPK) domain. Based on this domain structure GKs are anticipated to link G-protein and phospholipid signal pathways; however, their functions are currently unknown. Expression analyses of the 12 GK genes in Phytophthora infestans and their orthologues in Phytophthora sojae, revealed differential expression during asexual development. PiGK1 and PiGK4 were fused to monomeric red fluorescent protein (mRFP) and ectopically expressed in P.?infestans. In growing hyphae different subcellular distribution patterns were observed indicating that these two GKs act independently during development. We focused on the functional analyses of PiGK4. Its localization suggested involvement in cell differentiation and elongation and its 7-TM domain showed a canonical GPCR membrane topology. Silencing of GK4 and overexpression of full-length and truncated constructs in P.?infestans revealed that PiGK4 is not only involved in spore germination and hyphal elongation but also in sporangia cleavage and infection. PMID:23448716

Hua, Chenlei; Meijer, Harold J G; de Keijzer, Jeroen; Zhao, Wei; Wang, Yuanchao; Govers, Francine



Single-strand-conformation polymorphism of ribosomal DNA for rapid species differentiation in genus Phytophthora.  


Single-strand-conformation polymorphism (SSCP) of ribosomal DNA of 29 species (282 isolates) of Phytophthora was characterized in this study. Phytophthora boehmeriae, Phytophthora botryosa, Phytophthora cactorum, Phytophthora cambivora, Phytophthora capsici, Phytophthora cinnamomi, Phytophthora colocasiae, Phytophthora fragariae, Phytophthora heveae, Phytophthora hibernalis, Phytophthora ilicis, Phytophthora infestans, Phytophthora katsurae, Phytophthora lateralis, Phytophthora meadii, Phytophthora medicaginis, Phytophthora megakarya, Phytophthora nicotianae, Phytophthora palmivora, Phytophthora phaseoli, Phytophthora pseudotsugae, Phytophthora sojae, Phytophthora syringae, and Phytophthora tropicalis each showed a unique SSCP pattern. Phytophthora citricola, Phytophthora citrophthora, Phytophthora cryptogea, Phytophthora drechsleri, and Phytophthora megasperma each had more than one distinct pattern. A single-stranded DNA ladder also was developed, which facilitates comparison of SSCP patterns within and between gels. With a single DNA fingerprint, 277 isolates of Phytophthora recovered from irrigation water and plant tissues in Virginia were all correctly identified into eight species at substantially reduced time, labor, and cost. The SSCP analysis presented in this work will aid in studies on taxonomy, genetics, and ecology of the genus Phytophthora. PMID:12892637

Kong, Ping; Hong, Chuanxue; Richardson, Patricia A; Gallegly, Mannon E



Uncovering the salt response of soybean by unraveling its wild and cultivated functional genomes using tag sequencing.  


Soil salinity has very adverse effects on growth and yield of crop plants. Several salt tolerant wild accessions and cultivars are reported in soybean. Functional genomes of salt tolerant Glycine soja and a salt sensitive genotype of Glycine max were investigated to understand the mechanism of salt tolerance in soybean. For this purpose, four libraries were constructed for Tag sequencing on Illumina platform. We identify around 490 salt responsive genes which included a number of transcription factors, signaling proteins, translation factors and structural genes like transporters, multidrug resistance proteins, antiporters, chaperons, aquaporins etc. The gene expression levels and ratio of up/down-regulated genes was greater in tolerant plants. Translation related genes remained stable or showed slightly higher expression in tolerant plants under salinity stress. Further analyses of sequenced data and the annotations for gene ontology and pathways indicated that soybean adapts to salt stress through ABA biosynthesis and regulation of translation and signal transduction of structural genes. Manipulation of these pathways may mitigate the effect of salt stress thus enhancing salt tolerance. PMID:23209559

Ali, Zulfiqar; Zhang, Da Yong; Xu, Zhao Long; Xu, Ling; Yi, Jin Xin; He, Xiao Lan; Huang, Yi Hong; Liu, Xiao Qing; Khan, Asif Ali; Trethowan, Richard M; Ma, Hong Xiang



Anxiolytic Effects of Flavonoids in Animal Models of Posttraumatic Stress Disorder  

PubMed Central

The dysregulation of the serotonergic system has long been recognized as an important factor underlying the pathophysiology of PTSD. To date, SSRIs have already been established as the firstline pharmacotherapeutic agents for treating acute and chronic PTSD. However, SSRIs largely have several disadvantages which limit their utility. Our previous study has also shown that administration of the total flavonoids, isolated from the extract of Xiaobuxin-Tang (XBXT, mild mind-easing decoction), comprising four Chinese medicines including Haematitum, Flos Inulae, Folium Phyllostachydis Henonis, and Semen Sojae Preparatum, exerted significant antidepressant-like effect in chronically mildly stressed rats, possibly mediated by serotonergic activation. Since the central serotonergic dysfunction is an important and well-known cause mediating the pathophysiology of trauma-related symptoms in PTSD, it is reasonable to predict that flavonoids may exert therapeutic effects on PTSD in animal models. Therefore, the present study aims to examine the effect of flavonoids in alleviating the enhanced anxiety and fear response induced in two PTSD animal models. Ser, an SSRI, was administered as a positive control. Furthermore, the changes of brain monoaminergic neurotransmitters after chronic flavonoids administration have also been assessed in SPS-treated rats.

Zhang, Li-Ming; Yao, Jia-Zhi; Li, Yang; Li, Kai; Chen, Hong-Xia; Zhang, You-Zhi; Li, Yun-Feng



Novel alleles among soybean Bowman-Birk proteinase inhibitor gene families.  


Trypsin inhibitors have been found in various animals, plants and microorganisms. There were two types of trypsin inhibitors in soybean including Bowman-Birk protease inhibitors (BBI) and Kunitz inhibitors (KTI). The different BBI genes from wild soybean (G. soja) and cultivated soybean (G. max) formed a multigene family. We constructed a cDNA library of cultivar 'SuiNong 14' seed at the R7 growth stage using the SMART Kit. Seventeen contigs or singletons were highly homologous to soybean protease inhibitors. Contigs of 5, 35, 8 and 9 were highly homologous to BBI family members BBI-A1, BBI-A2, BBI-C and BBI-D, respectively. Sequence analyses showed there were novel allelic variations among the 4 BBI members in SuiNong 14. Based on the comparison of soybean seed cDNA libraries from different developmental stages, it was apparent that the expression of trypsin inhibitors increased during seed development in soybean. Phylogenetic analysis of BBI gene sequences among dicotyledonous and monocotyledonous plants demonstrated that these genes shared a common progenitor. PMID:18677596

Wang, Yueping; Chen, Xiongting; Qiu, Lijuan



Phylogenetic relationship of Lotus uliginosus symbionts with bradyrhizobia nodulating genistoid legumes.  


Lotus species are legumes with potential for pastures in soils with low-fertility and environmental constraints. The aim of this work was to characterize bacteria that establish efficient nitrogen-fixing symbiosis with the forage species Lotus uliginosus. A total of 39 isolates were obtained from nodules of L. uliginosus naturally growing in two different locations of Portugal. Molecular identification of the isolates plus the commercial inoculant strain NZP2039 was performed by REP-PCR, 16S rRNA RFLP, and 16S rRNA, glnII and recA sequence analyses. Limited genetic diversity was found among the L. uliginosus symbionts, which showed a close phylogenetic relationship with the species Bradyrhizobium japonicum. The symbiotic nifH, nodA and nodC gene sequences were closely related with the corresponding genes of various Bradyrhizobium strains isolated from Lupinus and other genistoid legumes and therefore were phylogenetically separated from other Lotus spp. rhizobia. The L. uliginosus bradyrhizobia were able to nodulate and fix nitrogen in association with L. uliginosus, could nodulate Lotus corniculatus with generally poor nitrogen-fixing efficiency, formed nonfixing nodules in Lotus tenuis and Lupinus luteus roots and were unable to nodulate Glycine soja or Glycine max. Thus, L. uliginosus rhizobia seem closely related to B. japonicum biovar genistearum strains. PMID:22092879

Lorite, María J; Videira e Castro, Isabel; Muñoz, Socorro; Sanjuán, Juan



Culture condition-dependent metabolite profiling of Aspergillus fumigatus with antifungal activity.  


Three sections of Aspergillus (five species, 21 strains) were classified according to culture medium-dependent and time-dependent secondary metabolite profile-based chemotaxonomy. Secondary metabolites were analysed by liquid chromatography-electrospray ionisation tandem mass spectrometry (LC-ESI-MS-MS) and multivariate statistical methods. From the Aspergillus sections that were cultured on malt extract agar (MEA) and Czapek yeast extract agar (CYA) for 7, 12, and 16 d, Aspergillus sections Fumigati (A. fumigatus), Nigri (A. niger), and Flavi (A. flavus, A. oryzae, and A. sojae) clustered separately on the basis of the results of the secondary metabolite analyses at 16 d regardless of culture medium. Based on orthogonal projection to latent structures discriminant analysis by partial least squares discriminant analysis (PLS-DA), we identified the secondary metabolites that helped differentiate sections between A. fumigatus and Aspergillus section Flavi to be gliotoxin G, fumigatin oxide, fumigatin, pseurotin A or D, fumiquinazoline D, fumagillin, helvolic acid, 1,2-dihydrohelvolic acid, and 5,8-dihydroxy-9,12-octadecadienoic acid (5,8-diHODE). Among these compounds, fumagillin, helvolic acid, and 1,2-dihydrohelvolic acid of A. fumigatus showed antifungal activities against Malassezia furfur, which is lipophilic yeast that causes epidermal skin disorders. PMID:23537878

Kang, Daejung; Son, Gun Hee; Park, Hye Min; Kim, Jiyoung; Choi, Jung Nam; Kim, Hyang Yeon; Lee, Sarah; Hong, Seung-Beom; Lee, Choong Hwan



Soybean glyceollins mitigate inducible nitric oxide synthase and cyclooxygenase-2 expression levels via suppression of the NF-?B signaling pathway in RAW 264.7 cells.  


Glyceollins, produced to induce disease resistance responses against specific species, such as an incompatible pathogen Phytophthora sojae in soybeans, have the potential to exhibit anti-inflammatory activity in RAW 264.7 cells. To investigate the anti-inflammatory effects of elicited glyceollins via a signaling pathway, we studied the glyceollin signaling pathway using several assays including RNA and protein expression levels. We found that soybean glyceollins significantly reduced LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production, as well as the expression of inducible ?? synthase (iNOS) and cyclooxygenase-2 (COX-2) via the suppression of NF-?B activation. Glyceollins also inhibited the phosphorylation of I?B? kinase (IKK), the degradation of I?B?, and the formation of NF-?B-DNA binding complex in a dose-dependent manner. Furthermore, they inhibited pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-?, interleukin (IL)-1? and IL-18, but increased the generation of the anti-inflammatory cytokine IL-10. Collectively, the present data show that glyceollins elicit potential anti-inflammatory effects by suppressing the NF-?B signaling pathway in RAW 264.7 cells. PMID:22246209

Yoon, Eun-Kyung; Kim, Hyun-Kyoung; Cui, Song; Kim, Yong-Hoon; Lee, Sang-Han



Molecular basis of recognition between phytophthora pathogens and their hosts.  


Recognition is the earliest step in any direct plant-microbe interaction. Recognition between Phytophthora pathogens, which are oomycetes, phylogenetically distinct from fungi, has been studied at two levels. Recognition of the host by the pathogen has focused on recognition of chemical, electrical, and physical features of plant roots by zoospores. Both host-specific factors such as isoflavones, and host-nonspecific factors such as amino acids, calcium, and electrical fields, influence zoospore taxis, encystment, cyst germination, and hyphal chemotropism in guiding the pathogen to potential infection sites. Recognition of the pathogen by the host defense machinery has been analyzed using biochemical and genetic approaches. Biochemical approaches have identified chemical elicitors of host defense responses, and in some cases, their cognate receptors from the host. Some elicitors, such as glucans and fatty acids, have broad host ranges, whereas others such as elicitins have narrow host ranges. Most elicitors identified appear to contribute primarily to basic or nonhost resistance. Genetic analysis has identified host resistance (R) genes and pathogen avirulence (Avr) genes that interact in a gene-for-gene manner. One Phytophthora Avr gene, Avr1b from P. sojae, has been cloned and characterized. It encodes a secreted elicitor that triggers a system-wide defense response in soybean plants carrying the cognate R gene, Rps1b. PMID:12147757

Tyler, Brett M



Overexpression of GsZFP1 enhances salt and drought tolerance in transgenic alfalfa (Medicago sativa L.).  


GsZFP1 encodes a Cys2/His2-type zinc-finger protein. In our previous study, when GsZFP1 was heterologously expressed in Arabidopsis, the transgenic Arabidopsis plants exhibited enhanced drought and cold tolerance. However, it is still unknown whether GsZFP1 is also involved in salt stress. GsZFP1 is from the wild legume Glycine soja. Therefore, the aims of this study were to further elucidate the functions of the GsZFP1 gene under salt and drought stress in the forage legume alfalfa and to investigate its biochemical and physiological functions under these stress conditions. Our data showed that overexpression of GsZFP1 in alfalfa resulted in enhanced salt tolerance. Under high salinity stress, greater relative membrane permeability and malondialdehyde (MDA) content were observed and more free proline and soluble sugars accumulated in transgenic alfalfa than in the wild-type (WT) plants; in addition, the transgenic lines accumulated less Na(+) and more K(+) in both the shoots and roots. Overexpression of GsZFP1 also enhanced the drought tolerance of alfalfa. The fold-inductions of stress-responsive marker gene expression, including MtCOR47, MtRAB18, MtP5CS, and MtRD2, were greater in transgenic alfalfa than those of WT under drought stress conditions. In conclusion, the transgenic alfalfa plants generated in this study could be used for farming in salt-affected as well as arid and semi-arid areas. PMID:23867600

Tang, Lili; Cai, Hua; Ji, Wei; Luo, Xiao; Wang, Zhenyu; Wu, Jing; Wang, Xuedong; Cui, Lin; Wang, Yang; Zhu, Yanming; Bai, Xi



Inactivation of duplicated nod factor receptor 5 (NFR5) genes in recessive loss-of-function non-nodulation mutants of allotetraploid soybean (Glycine max L. Merr.).  


Chemically induced non-nodulating nod139 and nn5 mutants of soybean (Glycine max) show no visible symptoms in response to rhizobial inoculation. Both exhibit recessive Mendelian inheritance suggesting loss of function. By allele determination and genetic complementation in nod139 and nn5, two highly related lipo-oligochitin LysM-type receptor kinase genes in Glycine max were cloned; they are presumed to be the critical nodulation-inducing (Nod) factor receptor similar to those of Lotus japonicus, pea and Medicago truncatula. These duplicated receptor genes were called GmNFR5alpha and GmNFR5beta. Nonsense mutations in GmNFR5alpha and GmNFR5beta were genetically complemented by both wild-type GmNFR5alpha and GmNFR5beta in transgenic roots, indicating that both genes are functional. Both genes lack introns. In cultivar Williams82 GmNFR5alpha is located in chromosome 11 and in tandem with GmLYK7 (a related LysM receptor kinase gene), while GmNFR5beta is in tandem with GmLYK4 in homologous chromosome 1, suggesting ancient synteny and regional segmental duplication. Both genes are wild type in G. soja CPI100070 and Harosoy63; however, a non-functional NFR5beta allele (NFR5beta*) was discovered in parental lines Bragg and Williams, which harbored an identical 1,407 bp retroelement-type insertion. This retroelement (GmRE-1) and related sequences are located in several soybean genome positions. Paradoxically, putatively unrelated soybean cultivars shared the same insertion, suggesting a smaller than anticipated genetic base in this crop. GmNFR5alpha but not GmNFR5beta* was expressed in inoculated and uninoculated tap and lateral root portions at about 10-25% of GmATS1 (ATP synthase subunit 1), but not in trifoliate leaves and shoot tips. PMID:20007291

Indrasumunar, Arief; Kereszt, Attila; Searle, Iain; Miyagi, Mikiko; Li, Dongxue; Nguyen, Cuc D T; Men, Artem; Carroll, Bernard J; Gresshoff, Peter M



Artificial selection for determinate growth habit in soybean.  


Determinacy is an agronomically important trait associated with the domestication in soybean (Glycine max). Most soybean cultivars are classifiable into indeterminate and determinate growth habit, whereas Glycine soja, the wild progenitor of soybean, is indeterminate. Indeterminate (Dt1/Dt1) and determinate (dt1/dt1) genotypes, when mated, produce progeny that segregate in a monogenic pattern. Here, we show evidence that Dt1 is a homolog (designated as GmTfl1) of Arabidopsis terminal flower 1 (TFL1), a regulatory gene encoding a signaling protein of shoot meristems. The transition from indeterminate to determinate phenotypes in soybean is associated with independent human selections of four distinct single-nucleotide substitutions in the GmTfl1 gene, each of which led to a single amino acid change. Genetic diversity of a minicore collection of Chinese soybean landraces assessed by simple sequence repeat (SSR) markers and allelic variation at the GmTfl1 locus suggest that human selection for determinacy took place at early stages of landrace radiation. The GmTfl1 allele introduced into a determinate-type (tfl1/tfl1) Arabidopsis mutants fully restored the wild-type (TFL1/TFL1) phenotype, but the Gmtfl1 allele in tfl1/tfl1 mutants did not result in apparent phenotypic change. These observations indicate that GmTfl1 complements the functions of TFL1 in Arabidopsis. However, the GmTfl1 homeolog, despite its more recent divergence from GmTfl1 than from Arabidopsis TFL1, appears to be sub- or neo-functionalized, as revealed by the differential expression of the two genes at multiple plant developmental stages and by allelic analysis at both loci. PMID:20421496

Tian, Zhixi; Wang, Xiaobo; Lee, Rian; Li, Yinghui; Specht, James E; Nelson, Randall L; McClean, Phillip E; Qiu, Lijuan; Ma, Jianxin



Nutrition rehabilitation of undernourished children utilizing Spiruline and Misola  

PubMed Central

Background Malnutrition constitutes a public health problem throughout the world and particularly in developing countries. Aims The objective of the study is to assess the impact of an elementary integrator composed of Spiruline (Spirulina platensis) and Misola (millet, soja, peanut) produced at the Centre Medical St Camille (CMSC) of Ouagadougou, Burkina Faso, on the nutritional status of undernourished children. Materials and methods 550 undernourished children of less than 5 years old were enrolled in this study, 455 showed severe marasma, 57 marasma of medium severity and 38 kwashiorkor plus marasma. We divided the children randomly into four groups: 170 were given Misola (731 ± 7 kcal/day), 170 were given Spiruline plus traditional meals (748 ± 6 kcal/day), 170 were given Spiruline plus Misola (767 ± 5 kcal/day). Forty children received only traditional meals (722 ± 8 kcal/day) and functioned as the control group. The duration of this study was eight weeks. Results and Discussion Anthropometrics and haematological parameters allowed us to appreciate both the nutritional and biological evolution of these children. The rehabilitation with Spiruline plus Misola (this association gave an energy intake of 767 ± 5 kcal/day with a protein assumption of 33.3 ± 1.2 g a day), both greater than Misola or Spiruline alone, seems to correct weight loss more quickly. Conclusion Our results indicate that Misola, Spiruline plus traditional meals or Spiruline plus Misola are all a good food supplement for undernourished children, but the rehabilitation by Spiruline plus Misola seems synergically favour the nutrition rehabilitation better than the simple addition of protein and energy intake.

Simpore, Jacques; Kabore, Fatoumata; Zongo, Frederic; Dansou, Deleli; Bere, Augustin; Pignatelli, Salvatore; Biondi, Daniela M; Ruberto, Giuseppe; Musumeci, Salvatore



Are algal genes in nonphotosynthetic protists evidence of historical plastid endosymbioses?  

PubMed Central

Background How photosynthetic organelles, or plastids, were acquired by diverse eukaryotes is among the most hotly debated topics in broad scale eukaryotic evolution. The history of plastid endosymbioses commonly is interpreted under the "chromalveolate" hypothesis, which requires numerous plastid losses from certain heterotrophic groups that now are entirely aplastidic. In this context, discoveries of putatively algal genes in plastid-lacking protists have been cited as evidence of gene transfer from a photosynthetic endosymbiont that subsequently was lost completely. Here we examine this evidence, as it pertains to the chromalveolate hypothesis, through genome-level statistical analyses of similarity scores from queries with two diatoms, Phaeodactylum tricornutum and Thalassiosira pseudonana, and two aplastidic sister taxa, Phytophthora ramorum and P. sojae. Results Contingency tests of specific predictions of the chromalveolate model find no evidence for an unusual red algal contribution to Phytophthora genomes, nor that putative cyanobacterial sequences that are present entered these genomes through a red algal endosymbiosis. Examination of genes unrelated to plastid function provide extraordinarily significant support for both of these predictions in diatoms, the control group where a red endosymbiosis is known to have occurred, but none of that support is present in genes specifically conserved between diatoms and oomycetes. In addition, we uncovered a strong association between overall sequence similarities among taxa and relative sizes of genomic data sets in numbers of genes. Conclusion Signal from "algal" genes in oomycete genomes is inconsistent with the chromalveolate hypothesis, and better explained by alternative models of sequence and genome evolution. Combined with the numerous sources of intragenomic phylogenetic conflict characterized previously, our results underscore the potential to be mislead by a posteriori interpretations of variable phylogenetic signals contained in complex genome-level data. They argue strongly for explicit testing of the different a priori assumptions inherent in competing evolutionary hypotheses.

Stiller, John W; Huang, Jinling; Ding, Qin; Tian, Jing; Goodwillie, Carol



Construction and characterization of a DNA probe for distinguishing strains of Aspergillus flavus.  


Repetitive DNA sequences have proven useful and reliable characters in evaluating genetic relatedness of strains at different levels of taxonomic classification. A DNA probe was constructed to distinguish among strains of Aspergillus flavus by DNA fingerprinting techniques. Chromosomal DNA of A. flavus var. flavus NRRL 6541 was partially digested with EcoRI and ligated to a Lambda Dash bacteriophage vector. Four lambda clones were identified which displayed multiple and distinct bands when hybridized with chromosomal DNA from seven strains of A. flavus var. flavus digested with either EcoRI or PstI. One of these clones was chosen for further analysis and was subcloned into pUC19. The subclone, pAF28, contained a 6.2-kb chromosomal DNA insert and was able to distinguish among strains characterized by K. E. Papa (Mycologia 78:98-101, 1986) as belonging to 22 different vegetative compatibility groups. The subclone identified unique banding patterns when hybridized to genomic DNA digested with PstI. The cloned probe may be species specific as it hybridized with the DNA of all isolates of A. flavus tested in addition to strains recognized as varieties of A. flavus (e.g., A. flavus var. oryzae, A. flavus var. parasiticus, and A. flavus var. sojae). pAF28 hybridized to a single band on a Southern blot with Aspergillus nomius DNA but did not hybridize with the DNA of other fungal species tested including Aspergillus ochraceus, Aspergillus auricomus, Aspergillus alliaceus, Fusarium moniliforme, and Penicillium thomii. PMID:7793909

McAlpin, C E; Mannarelli, B



A second eukaryotic group with mitochondrion-encoded tmRNA: in silico identification and experimental confirmation.  


In bacteria, stalled ribosomes are rescued by transfer-mRNA (tmRNA) that catalyzes two steps. First, a non-encoded alanine is added to the incomplete polypeptide chain by the tRNA (Ala) -like portion of tmRNA, and second, the ribosome switches to the mRNA-like domain of tmRNA, thus resuming protein synthesis. Mitochondrial DNA (mtDNA)-encoded mt-tmRNA is so far only known from jakobid protists, but we posit that the corresponding ssrA gene may also reside in other mtDNAs. Here we present a highly sensitive covariance model built from jakobid ssrA genes that identifies previously unrecognized ssrA homologs in mtDNAs of oomycetes. These genes, located in previously unassigned genomic regions, are circular permuted as in ?-Protobacteria, implying that pre-tmRNA is processed and the two pieces are held together by non-covalent interactions. RNA-Seq data from Phytophthora sojae confirm predicted processing sites as well as post-transcriptional addition of 3' CCA, a prerequisite for tmRNAs to be charged with alanine by alanyl-tRNA synthetase. Structure modeling of oomycete tmRNAs infers that the mRNA-like domain is lacking as in jakobids. Features of mitochondrial tmRNAs include the G-U pair at position three of the acceptor stem, a hallmark of bacterial tmRNAs, and a T-loop sequence that differs from that of standard tRNAs and most bacterial tmRNAs, forming alternative, virtually isosteric tertiary interactions with the D-loop. The anticodon stem has two additional G-A base pairs formed between the D-loop and the variable region, shortening the length of the variable region to a single nucleotide. PMID:23823571

Hafez, Mohamed; Burger, Gertraud; Steinberg, Sergey V; Lang, B Franz



GsTIFY10, a novel positive regulator of plant tolerance to bicarbonate stress and a repressor of jasmonate signaling.  


Recent discoveries show that TIFY family genes are plant-specific genes involved in the response to several abiotic stresses, also acting as key regulators of jasmonate signaling in Arabidopsis thaliana. However, there is limited information about this gene family in wild soybean, nor is its role in plant bicarbonate stress adaptation completely understood. Here, we isolated and characterized a novel TIFY family gene, GsTIFY10, from Glycine soja. GsTIFY10 could be induced by bicarbonate, salinity stress and the phytohormone JA, both in the leaves and roots of wild soybean. Over-expression of GsTIFY10 in Arabidopsis resulted in enhanced plant tolerance to bicarbonate stress during seed germination, early seedling and adult seedling developmental stages, and the expression levels of some bicarbonate stress response and stress-inducible marker genes were significantly higher in the GsTIFY10 overexpression lines than in wild-type plants. It was also found that GsTIFY10 could repress JA signal transduction. The roots of plants overexpressing GsTIFY10 grew longer than wild-type in the presence of MeJA, and some JA response and JA biosynthesis marker genes were suppressed in the GsTIFY10 overexpression lines. Subcellular localization studies using a GFP fusion protein showed that GsTIFY10 is localized to the nucleus. These results suggest that the newly isolated wild soybean GsTIFY10 is a positive regulator of plant bicarbonate stress tolerance and is also a repressor of jasmonate signaling, from hormone perception to transcriptional activity. PMID:21805375

Zhu, Dan; Bai, Xi; Chen, Chao; Chen, Qin; Cai, Hua; Li, Yong; Ji, Wei; Zhai, Hong; Lv, Dekang; Luo, Xiao; Zhu, Yanming



A single origin and moderate bottleneck during domestication of soybean (Glycine max): implications from microsatellites and nucleotide sequences  

PubMed Central

Background and Aims It is essential to illuminate the evolutionary history of crop domestication in order to understand further the origin and development of modern cultivation and agronomy; however, despite being one of the most important crops, the domestication origin and bottleneck of soybean (Glycine max) are poorly understood. In the present study, microsatellites and nucleotide sequences were employed to elucidate the domestication genetics of soybean. Methods The genomes of 79 landrace soybeans (endemic cultivated soybeans) and 231 wild soybeans (G. soja) that represented the species-wide distribution of wild soybean in East Asia were scanned with 56 microsatellites to identify the genetic structure and domestication origin of soybean. To understand better the domestication bottleneck, four nucleotide sequences were selected to simulate the domestication bottleneck. Key Results Model-based analysis revealed that most of the landrace genotypes were assigned to the inferred wild soybean cluster of south China, South Korea and Japan. Phylogeny for wild and landrace soybeans showed that all landrace soybeans formed a single cluster supporting a monophyletic origin of all the cultivars. The populations of the nearest branches which were basal to the cultivar lineage were wild soybeans from south China. The coalescent simulation detected a bottleneck severity of K? = 2 during soybean domestication, which could be explained by a foundation population of 6000 individuals if domestication duration lasted 3000 years. Conclusions As a result of integrating geographic distribution with microsatellite genotype assignment and phylogeny between landrace and wild soybeans, a single origin of soybean in south China is proposed. The coalescent simulation revealed a moderate genetic bottleneck with an effective wild soybean population used for domestication estimated to be ?2 % of the total number of ancestral wild soybeans. Wild soybeans in Asia, especially in south China contain tremendous genetic resources for cultivar improvement.

Guo, Juan; Wang, Yunsheng; Song, Chi; Zhou, Jianfeng; Qiu, Lijuan; Huang, Hongwen; Wang, Ying



Expression of wild soybean WRKY20 in Arabidopsis enhances drought tolerance and regulates ABA signalling.  


The WRKY-type transcription factors are involved in plant development and stress responses, but how the regulation of stress tolerance is related to plant development is largely unknown. GsWRKY20 was initially identified as a stress response gene using large-scale Glycine soja microarrays. Quantitative reverse transcription-PCR (qRT-PCR) showed that the expression of this gene was induced by abscisic acid (ABA), salt, cold, and drought. Overexpression of GsWRKY20 in Arabidopsis resulted in a decreased sensitivity to ABA during seed germination and early seedling growth. However, compared with the wild type, GsWRKY20 overexpression lines were more sensitive to ABA in stomatal closure, and exhibited a greater tolerance to drought stress, a decreased water loss rate, and a decreased stomatal density. Moreover, microarray and qRT-PCR assays showed that GsWRKY20 mediated ABA signalling by promoting the expression of negative regulators of ABA signalling, such as AtWRKY40, ABI1, and ABI2, while repressing the expression of the positive regulators of ABA, for example ABI5, ABI4, and ABF4. Interestingly, GsWRKY20 also positively regulates the expression of a group of wax biosynthetic genes. Further, evidence is provided to support that GsWRKY20 overexpression lines have more epicuticular wax crystals and a much thicker cuticle, which contribute to less chlorophyll leaching compared with the wild type. Taken together, the findings reveal an important role for GsWRKY20 in enhancing drought tolerance and regulating ABA signalling. PMID:23606412

Luo, Xiao; Bai, Xi; Sun, Xiaoli; Zhu, Dan; Liu, Baohui; Ji, Wei; Cai, Hua; Cao, Lei; Wu, Jing; Hu, Mengran; Liu, Xin; Tang, Lili; Zhu, Yanming



Mosaic origin of the heme biosynthesis pathway in photosynthetic eukaryotes.  


Heme biosynthesis represents one of the most essential metabolic pathways in living organisms, providing the precursors for cytochrome prosthetic groups, photosynthetic pigments, and vitamin B(12). Using genomic data, we have compared the heme pathway in the diatom Thalassiosira pseudonana and the red alga Cyanidioschyzon merolae to those of green algae and higher plants, as well as to those of heterotrophic eukaryotes (fungi, apicomplexans, and animals). Phylogenetic analyses showed the mosaic character of this pathway in photosynthetic eukaryotes. Although most of the algal and plant enzymes showed the expected plastid (cyanobacterial) origin, at least one of them (porphobilinogen deaminase) appears to have a mitochondrial (alpha-proteobacterial) origin. Another enzyme, glutamyl-tRNA synthase, obviously originated in the eukaryotic nucleus. Because all the plastid-targeted sequences consistently form a well-supported cluster, this suggests that genes were either transferred from the primary endosymbiont (cyanobacteria) to the primary host nucleus shortly after the primary endosymbiotic event or replaced with genes from other sources at an equally early time, i.e., before the formation of three primary plastid lineages. The one striking exception to this pattern is ferrochelatase, the enzyme catalyzing the first committed step to heme and bilin pigments. In this case, two red algal sequences do not cluster either with the other plastid sequences or with cyanobacterial sequences and appear to have a proteobacterial origin like that of the apicomplexan parasites Plasmodium and Toxoplasma. Although the heterokonts also acquired their plastid via secondary endosymbiosis from a red alga, the diatom has a typical plastid-cyanobacterial ferrochelatase. We have not found any remnants of the plastidlike heme pathway in the nonphotosynthetic heterokonts Phytophthora ramorum and Phytophthora sojae. PMID:16093570

Oborník, Miroslav; Green, Beverley R



Bradyrhizobium huanghuaihaiense sp. nov., an effective symbiotic bacterium isolated from soybean (Glycine max L.) nodules.  


In a survey of the biodiversity and biogeography of rhizobia associated with soybean (Glycine max L.) in different sites of the Northern (Huang-Huai-Hai) Plain of China, ten strains were defined as representing a novel genomic species in the genus of Bradyrhizobium. They were distinguished from defined species in restriction fragment length polymorphism (RFLP) analysis of the 16S rRNA gene and the 16S-23S rRNA gene intergenic spacer (IGS). In BOX-PCR, these strains presented two patterns that shared 94% similarity, demonstrating that they were a homogenous group with limited diversity. In phylogenetic analyses of the 16S rRNA gene, IGS and housekeeping gene sequences, four representative strains formed a distant lineage within the genus Bradyrhizobium, which was consistent with the results of DNA-DNA hybridization. The strains of this novel group formed effective nodules with G. max, Glycine soja and Vigna unguiculata in cross-nodulation tests and harboured symbiotic genes (nodC and nifH) identical to those of reference strains of Bradyrhizobium japonicum, Bradyrhizobium liaoningense and 'Bradyrhizobium daqingense' originating from soybean, implying that the novel group may have obtained these symbiotic genes by lateral gene transfer. In analyses of cellular fatty acids and phenotypic features, some differences were found between the novel group and related Bradyrhizobium species, demonstrating that the novel group is distinct phenotypically from other Bradyrhizobium species. Based upon the data obtained, these strains are proposed to represent a novel species, Bradyrhizobium huanghuaihaiense sp. nov., with CCBAU 23303(T) (?=?LMG 26136(T) ?=?CGMCC 1.10948(T) ?=?HAMBI 3180(T)) as the type strain. The DNA G+C content of strain CCBAU 23303(T) is 61.5 mol% (T(m)). PMID:22003042

Zhang, Yan Ming; Li, Ying; Chen, Wen Feng; Wang, En Tao; Sui, Xin Hua; Li, Qin Qin; Zhang, Yun Zeng; Zhou, Yu Guang; Chen, Wen Xin



Survey and analysis of microsatellites from transcript sequences in Phytophthora species: frequency, distribution, and potential as markers for the genus  

PubMed Central

Background Members of the genus Phytophthora are notorious pathogens with world-wide distribution. The most devastating species include P. infestans, P. ramorum and P. sojae. In order to develop molecular methods for routinely characterizing their populations and to gain a better insight into the organization and evolution of their genomes, we used an in silico approach to survey and compare simple sequence repeats (SSRs) in transcript sequences from these three species. We compared the occurrence, relative abundance, relative density and cross-species transferability of the SSRs in these oomycetes. Results The number of SSRs in oomycetes transcribed sequences is low and long SSRs are rare. The in silico transferability of SSRs among the Phytophthora species was analyzed for all sets generated, and primers were selected on the basis of similarity as possible candidates for transferability to other Phytophthora species. Sequences encoding putative pathogenicity factors from all three Phytophthora species were also surveyed for presence of SSRs. However, no correlation between gene function and SSR abundance was observed. The SSR survey results, and the primer pairs designed for all SSRs from the three species, were deposited in a public database. Conclusion In all cases the most common SSRs were trinucleotide repeat units with low repeat numbers. A proportion (7.5%) of primers could be transferred with 90% similarity between at least two species of Phytophthora. This information represents a valuable source of molecular markers for use in population genetics, genetic mapping and strain fingerprinting studies of oomycetes, and illustrates how genomic databases can be exploited to generate data-mining filters for SSRs before experimental validation.

Garnica, Diana P; Pinzon, Andres M; Quesada-Ocampo, Lina M; Bernal, Adriana J; Barreto, Emiliano; Grunwald, Niklaus J; Restrepo, Silvia



Multiple Horizontal Gene Transfer Events and Domain Fusions Have Created Novel Regulatory and Metabolic Networks in the Oomycete Genome  

PubMed Central

Complex enzymes with multiple catalytic activities are hypothesized to have evolved from more primitive precursors. Global analysis of the Phytophthora sojae genome using conservative criteria for evaluation of complex proteins identified 273 novel multifunctional proteins that were also conserved in P. ramorum. Each of these proteins contains combinations of protein motifs that are not present in bacterial, plant, animal, or fungal genomes. A subset of these proteins were also identified in the two diatom genomes, but the majority of these proteins have formed after the split between diatoms and oomycetes. Documentation of multiple cases of domain fusions that are common to both oomycetes and diatom genomes lends additional support for the hypothesis that oomycetes and diatoms are monophyletic. Bifunctional proteins that catalyze two steps in a metabolic pathway can be used to infer the interaction of orthologous proteins that exist as separate entities in other genomes. We postulated that the novel multifunctional proteins of oomycetes could function as potential Rosetta Stones to identify interacting proteins of conserved metabolic and regulatory networks in other eukaryotic genomes. However ortholog analysis of each domain within our set of 273 multifunctional proteins against 39 sequenced bacterial and eukaryotic genomes, identified only 18 candidate Rosetta Stone proteins. Thus the majority of multifunctional proteins are not Rosetta Stones, but they may nonetheless be useful in identifying novel metabolic and regulatory networks in oomycetes. Phylogenetic analysis of all the enzymes in three pathways with one or more novel multifunctional proteins was conducted to determine the probable origins of individual enzymes. These analyses revealed multiple examples of horizontal transfer from both bacterial genomes and the photosynthetic endosymbiont in the ancestral genome of Stramenopiles. The complexity of the phylogenetic origins of these metabolic pathways and the paucity of Rosetta Stones relative to the total number of multifunctional proteins suggests that the proteome of oomycetes has few features in common with other Kingdoms.

Morris, Paul Francis; Schlosser, Laura Rose; Onasch, Katherine Diane; Wittenschlaeger, Tom; Austin, Ryan; Provart, Nicholas



Comparative genomic analysis of soybean flowering genes.  


Flowering is an important agronomic trait that determines crop yield. Soybean is a major oilseed legume crop used for human and animal feed. Legumes have unique vegetative and floral complexities. Our understanding of the molecular basis of flower initiation and development in legumes is limited. Here, we address this by using a computational approach to examine flowering regulatory genes in the soybean genome in comparison to the most studied model plant, Arabidopsis. For this comparison, a genome-wide analysis of orthologue groups was performed, followed by an in silico gene expression analysis of the identified soybean flowering genes. Phylogenetic analyses of the gene families highlighted the evolutionary relationships among these candidates. Our study identified key flowering genes in soybean and indicates that the vernalisation and the ambient-temperature pathways seem to be the most variant in soybean. A comparison of the orthologue groups containing flowering genes indicated that, on average, each Arabidopsis flowering gene has 2-3 orthologous copies in soybean. Our analysis highlighted that the CDF3, VRN1, SVP, AP3 and PIF3 genes are paralogue-rich genes in soybean. Furthermore, the genome mapping of the soybean flowering genes showed that these genes are scattered randomly across the genome. A paralogue comparison indicated that the soybean genes comprising the largest orthologue group are clustered in a 1.4 Mb region on chromosome 16 of soybean. Furthermore, a comparison with the undomesticated soybean (Glycine soja) revealed that there are hundreds of SNPs that are associated with putative soybean flowering genes and that there are structural variants that may affect the genes of the light-signalling and ambient-temperature pathways in soybean. Our study provides a framework for the soybean flowering pathway and insights into the relationship and evolution of flowering genes between a short-day soybean and the long-day plant, Arabidopsis. PMID:22679494

Jung, Chol-Hee; Wong, Chui E; Singh, Mohan B; Bhalla, Prem L



Development and evaluation of SoySNP50K, a high-density genotyping array for soybean.  


The objective of this research was to identify single nucleotide polymorphisms (SNPs) and to develop an Illumina Infinium BeadChip that contained over 50,000 SNPs from soybean (Glycine max L. Merr.). A total of 498,921,777 reads 35-45 bp in length were obtained from DNA sequence analysis of reduced representation libraries from several soybean accessions which included six cultivated and two wild soybean (G. soja Sieb. et Zucc.) genotypes. These reads were mapped to the soybean whole genome sequence and 209,903 SNPs were identified. After applying several filters, a total of 146,161 of the 209,903 SNPs were determined to be ideal candidates for Illumina Infinium II BeadChip design. To equalize the distance between selected SNPs, increase assay success rate, and minimize the number of SNPs with low minor allele frequency, an iteration algorithm based on a selection index was developed and used to select 60,800 SNPs for Infinium BeadChip design. Of the 60,800 SNPs, 50,701 were targeted to euchromatic regions and 10,000 to heterochromatic regions of the 20 soybean chromosomes. In addition, 99 SNPs were targeted to unanchored sequence scaffolds. Of the 60,800 SNPs, a total of 52,041 passed Illumina's manufacturing phase to produce the SoySNP50K iSelect BeadChip. Validation of the SoySNP50K chip with 96 landrace genotypes, 96 elite cultivars and 96 wild soybean accessions showed that 47,337 SNPs were polymorphic and generated successful SNP allele calls. In addition, 40,841 of the 47,337 SNPs (86%) had minor allele frequencies ? 10% among the landraces, elite cultivars and the wild soybean accessions. A total of 620 and 42 candidate regions which may be associated with domestication and recent selection were identified, respectively. The SoySNP50K iSelect SNP beadchip will be a powerful tool for characterizing soybean genetic diversity and linkage disequilibrium, and for constructing high resolution linkage maps to improve the soybean whole genome sequence assembly. PMID:23372807

Song, Qijian; Hyten, David L; Jia, Gaofeng; Quigley, Charles V; Fickus, Edward W; Nelson, Randall L; Cregan, Perry B



The kinome of Phytophthora infestans reveals oomycete-specific innovations and links to other taxonomic groups  

PubMed Central

Background Oomycetes are a large group of economically and ecologically important species. Its most notorious member is Phytophthora infestans, the cause of the devastating potato late blight disease. The life cycle of P. infestans involves hyphae which differentiate into spores used for dispersal and host infection. Protein phosphorylation likely plays crucial roles in these stages, and to help understand this we present here a genome-wide analysis of the protein kinases of P. infestans and several relatives. The study also provides new insight into kinase evolution since oomycetes are taxonomically distant from organisms with well-characterized kinomes. Results Bioinformatic searches of the genomes of P. infestans, P. ramorum, and P. sojae reveal they have similar kinomes, which for P. infestans contains 354 eukaryotic protein kinases (ePKs) and 18 atypical kinases (aPKs), equaling 2% of total genes. After refining gene models, most were classifiable into families seen in other eukaryotes. Some ePK families are nevertheless unusual, especially the tyrosine kinase-like (TKL) group which includes large oomycete-specific subfamilies. Also identified were two tyrosine kinases, which are rare in non-metazoans. Several ePKs bear accessory domains not identified previously on kinases, such as cyclin-dependent kinases with integral cyclin domains. Most ePKs lack accessory domains, implying that many are regulated transcriptionally. This was confirmed by mRNA expression-profiling studies that showed that two-thirds vary significantly between hyphae, sporangia, and zoospores. Comparisons to neighboring taxa (apicomplexans, ciliates, diatoms) revealed both clade-specific and conserved features, and multiple connections to plant kinases were observed. The kinome of Hyaloperonospora arabidopsidis, an oomycete with a simpler life cycle than P. infestans, was found to be one-third smaller. Some differences may be attributable to gene clustering, which facilitates subfamily expansion (or loss) through unequal crossing-over. Conclusion The large sizes of the Phytophthora kinomes imply that phosphorylation plays major roles in their life cycles. Their kinomes also include many novel ePKs, some specific to oomycetes or shared with neighboring groups. Little experimentation to date has addressed the biological functions of oomycete kinases, but this should be stimulated by the structural, evolutionary, and expression data presented here. This may lead to targets for disease control.



Three-dimensional geometry and tectonostratigraphy of the Pennine zone, Central Alps, Switzerland and Northern Italy  

NASA Astrophysics Data System (ADS)

Continental collision during Alpine orogenesis entailed a polyphase deformation history (D 1-D 5) in the Pennine zone of the Central Alps. The regional tectonostratigraphy was basically developed during D 1 and D 2, characterised by isoclinal, typically north-closing recumbent anticlines, separated by pinched-in synclines, on the scale of tens of kilometres. Later deformation phases (D 3 and D 4) warped the stack into wavy to open folds. Exhumation of this zone resulted locally in later vertical shortening and folding of already steep fabrics (D 5). Three-dimensional models of the nappe pile were constructed, based on geostatistical assessment of the regional foliation field and considering the abundant structural field data. These models indicate the existence of five principal tectonostratigraphic levels developed during D 1 and thus equivalent to nappe units s. str.: the Gotthard, the Leventina-Antigorio, the Maggia-Simano (and probably the Monte Leone as well as the Composite Lepontine Series), Lebendun-Soja and Adula-Cima Lunga levels. All these tectonic units formed part of the passive continental margin of Europe prior to the onset of the Alpine orogenesis. Individual isoclinal post-nappe folds reflect relative displacements on the order of 40 km or more. The most prominent D 2 post-nappe structure is the Wandfluhhorn Fold, structurally equivalent to the northern closure of the Leventina-Lucomagno Antiform. The Lebendun and Monte Leone folds are of similar magnitudes and also affect the whole nappe pile, whereas the smaller Mogno and Molare synforms only refold the Maggia-Simano nappe internally. Principal D 3 and D 4 structures are the tight Mergoscia Synform directly north of the Insubric Fault between Bellinzona and Locarno (Southern Steep Belt), the Maggia Steep Zone, forming the steep western limb of the Campo Tencia Synform and subdividing the Lepontine dome into the Simplon and Ticino subdomes, the Chiéra Synform steepening the dominant foliation in the north (Northern Steep Belt) and the Vanzone and Claro Antiforms steepening the dominant foliation in the south (Southern Steep Belt). The current geometry of the Northern and Southern Steep Belts reflects an interplay between D 4 and D 3, involving both fold interference and reactivation/tightening.

Maxelon, Michael; Mancktelow, Neil S.



RPG1-B-Derived Resistance to AvrB-Expressing Pseudomonas syringae Requires RIN4-Like Proteins in Soybean1[W][OA  

PubMed Central

Soybean (Glycine max) RPG1-B (for resistance to Pseudomonas syringae pv glycinea) mediates species-specific resistance to P. syringae expressing the avirulence protein AvrB, similar to the nonorthologous RPM1 in Arabidopsis (Arabidopsis thaliana). RPM1-derived signaling is presumably induced upon AvrB-derived modification of the RPM1-interacting protein, RIN4 (for RPM1-interacting 4). We show that, similar to RPM1, RPG1-B does not directly interact with AvrB but associates with RIN4-like proteins from soybean. Unlike Arabidopsis, soybean contains at least four RIN4-like proteins (GmRIN4a to GmRIN4d). GmRIN4b, but not GmRIN4a, complements the Arabidopsis rin4 mutation. Both GmRIN4a and GmRIN4b bind AvrB, but only GmRIN4b binds RPG1-B. Silencing either GmRIN4a or GmRIN4b abrogates RPG1-B-derived resistance to P. syringae expressing AvrB. Binding studies show that GmRIN4b interacts with GmRIN4a as well as with two other AvrB/RPG1-B-interacting isoforms, GmRIN4c and GmRIN4d. The lack of functional redundancy among GmRIN4a and GmRIN4b and their abilities to interact with each other suggest that the two proteins might function as a heteromeric complex in mediating RPG1-B-derived resistance. Silencing GmRIN4a or GmRIN4b in rpg1-b plants enhances basal resistance to virulent strains of P. syringae and the oomycete Phytophthora sojae. Interestingly, GmRIN4a- or GmRIN4b-silenced rpg1-b plants respond differently to AvrB-expressing bacteria. Although both GmRIN4a and GmRIN4b function to monitor AvrB in the presence of RPG1-B, GmRIN4a, but not GmRIN4b, negatively regulates AvrB virulence activity in the absence of RPG1-B.

Selote, Devarshi; Kachroo, Aardra



Basaltic Martian analogues from the Baikal Rift Zone and Mongolian terranes  

NASA Astrophysics Data System (ADS)

In order to compare the results of studies of the western part of the Valles Marineris canyon on Mars there have been done field works on terrestrial surface areas similar with regard to geological setting and environmental conditions. One of the possible terrestrial analogues of the Valles Marineris canyon is the Baikal Rift Zone [1]. Field investigations have been done on the south end of the Baikal Lake, in the Khamar-Daban massif, where the outcrops of volcanic rocks occur. The second part of the field works has been done in the Mongolian terranes: Mandalovoo, Gobi Altay and Bayanhongor, because of environmental conditions being similar to those on Mars. The Mandalovoo terrane comprises a nearly continuous Paleozoic islandarc sequence [2]. In the Gobi Altay terrane an older sequence is capped by younger Devonian-Triassic volcanic-sedimentary deposits [2]. The Bayanhongor terrane forms a northwest-trending, discontinuous, narrow belt that consists of a large ophiolite allochton [3]. The collected samples of basalts derive from various geologic environments. The CORONA satellite-images have been used for the imaging of the Khamar-Daban massif and the Mandalovoo terrane. These images have the same spatial resolution and range as the Mars Orbiter Camera images of the Mars Global Surveyor mission. In the Mandalovoo terrane these images allowed to find an area with large amounts of tectonic structures, mainly faults (part of the Ongi massif), similar to the studied area on Mars. Microscopic observations in thin sections show diversification of composition and structures of basalts. These rocks have mostly a porphyric structure, rarely aphyric. The main components are plagioclases, pyroxenes and olivines phenocrysts, in different proportions. The groundmass usually consist of plagioclases, pyroxenes and opaques. The most diversified are basalts from the Mandalovoo terrane. Infrared spectroscopy has been used to analyse the composition of the rock material and compare these results with those of Martian missions. The range of the recorded spectra is from 400 to 2000 cm-1. Characteristic absorption bands in the spectra confirm the presence of minerals observed in thin sections and moreover, indicate the presence of leucite, analcime and saponite. The analyzed spectra have been compared with those of the Planetary Fourier Spectrometer of the Mars Express mission. The long wavelength channel of this spectrometer has a similar spectral range, from 230 to 2000 cm-1. This comparison indicates the similar absorption bands of clinopyroxenes in both spectra. References: [1] Komatsu G. (2003) Lunar Planet. Sci. XXXIV, abstract 1314. [2] Soja C. M. and White B. (2006) Geol. Soc. Amer., 38, 90. [3] Parfenov L. M. et al. (2002) Northeast Asia geodynamics map.

Gurgurewicz, J.; Kostylew, J.



Fullerene-C60 incorporated in liposome exerts persistent hydroxyl radical-scavenging activity and cytoprotection in UVA/B-irradiated keratinocytes.  


The aim of this study is to examine antioxidant activity of fullerene-C60 (C60) incorporated in liposome (LpsmFlln, a diameter of 75.6 nm). LpsmFlln is water-soluble, and composed of hydrogenated lecithin of 89.7%, glycine soja sterol of 10% and C60 of 0.3%. Hydroxyl radicals (*OH), generated from UVA- or UVB-irradiated H2O2, were scavenged by LpsmFlln but not by C60-lacking Lpsm as assessed by ESR, showing that the active principle is C60 as scanty as 1/415 weight versus LpsmFlln; the *OH amount (% of non-additive control) was decreased, LpsmFlln-dose-dependently, and for 0.5% LpsmFlln (C60-eq.:16.7 microM) to 34.1% or 78.3% upon irradiation with UVA (12 J/cm2) or UVB (500 mJ/cm2), respectively, showing the superiority for UVA to UVB in terms of the *OH scavenging of LpsmFlln. Cell viability of human skin keratinocytes HaCaT decreased to 41.1% upon UVA-irradiation at 10 J/cm2, but retained to 60.6% with 0.025% LpsmFlln (C60-eq.: 0.84 microM) together with prevention of cell-morphological degeneration, in contrast to scarce effects of C60-lacking Lpsm. The scavenging activity for Fenton reaction-generated *OH, detected by DMPO/ESR, was 96.2% or 72.2% (% of no-additive control) at 1 min and decreased time-dependently to 24.8% or 28.3% at 12 min with 16.7 microM L-ascorbic acid (Asc) or Trolox, respectively, whereas 0.5% LpsmFlln (C60-eq:16.7 microM, the same concentration as for Asc) diminished *OH by 90.9% at 1 min and 91.5% even at 12 min, demonstrating the superiority of LpsmFlln to Asc or Trolox in terms of persistence of *OH-scavenging ability. Repressive efficacy on beta-carotene discoloration (% of control) for 60 min was in the order, based on the same molar or weight concentration: 1.3%:3.34 microM Asc < 25.0%:0.1% Lpsm < 36.3%:0.1% LpsmFlln (C60-eq.:3.34 microM) < 57.2%:3.34 microM Trolox, indicating the preventive effect of LpsmFlln against beta-carotene oxidation. Thus, LpsmFlln was demonstrated for an antioxidant ability characteristic of long-term persistence, and is attributed to fullerene-C60 but scarcely to Lpsm in all the tests examined, and is expected as the skin-protecting agent against oxidative stress. PMID:21780373

Kato, Shinya; Aoshima, Hisae; Saitoh, Yasukazu; Miwa, Nobuhiko



Potential of Spirulina Platensis as a Nutritional Supplement in Malnourished HIV-Infected Adults in Sub-Saharan Africa: A Randomised, Single-Blind Study  

PubMed Central

Background: Malnutrition is a major global public health issue and its impact on communities and individuals is more dramatic in Sub-Saharan Africa, where it is compounded by widespread poverty and generalized high prevalence of human immunodeficiency virus (HIV). Therefore, malnutrition should be addressed through a multisectorial approach, and malnourished individuals should have access to nutritional rehabilitation molecules that are affordable, accessible, rich in nutrient and efficient. We thus assessed the efficacy of two affordable and accessible nutritional supplements, spirulina platensis versus soya beans among malnourished HIV-infected adults. Methods: Undernourished patients, naïve of, but eligible to antiretroviral treatment (ART), aged 18 to 35 years were enrolled and randomly assigned to two groups. The first group received spirulina (Group A) as food supplement and the second received soya beans (Group B). Patients were initiated ART simultaneously with supplements. Food supplements were auto-administered daily, the quantity being calculated according to weight to provide 1.5 g/kg body weight of proteins with 25% from supplements (spirulina and soya beans). Patients were monitored at baseline and followed-up during twelve weeks for anthropometric parameters, body composition, haemoglobin and serum albumin, CD4 count and viral load. Results: Fifty-two patients were enrolled (Group A: 26 and Group B: 26). The mean age was 26.4 ± 4.9 years (Group A) and 28.7 ± 4.8 (Group B) with no significant difference between groups (P = 0.10). After 12 weeks, weight and BMI significantly improved in both groups (P < 0.001 within each group). The mean gain in weight and BMI in Group A and B were 4.8 vs. 6.5 kg, (P = 0.68) and 1.3 vs. 1.90 Kg/m2, (P = 0.82) respectively. In terms of body composition, fat free mass (FFM) did not significantly increase within each group (40.5 vs. 42.2 Kg, P = 0.56 for Group A; 39.2 vs. 39.0 Kg, P = 0.22 for Group B). But when compared between the two groups at the end of the trial, FFM was significantly higher in the spirulina group (42.2 vs. 39.0 Kg, P = 0.01). The haemoglobin level rose significantly within groups (P < 0.001 for each group) with no difference between groups (P = 0.77). Serum albumin level did not increase significantly within groups (P < 0.90 vs. P < 0.82) with no difference between groups (P = 0.39). The increase in CD4 cell count within groups was significant (P < 0.01 in both groups), with a significantly higher CD4 count in the spirulina group compared to subjects on soya beans at the end of the study (P = 0.02). Within each group, HIV viral load significantly reduced at the end of the study (P < 0.001 and P = 0.04 for spirulina and soya beans groups respectively). Between the groups, the viral load was similar at baseline but significantly reduced in the spirulina group at the end of the study (P = 0.02). Conclusion: We therefore conclude in this preliminary study, firstly, that both spirulina and soja improve on nutritional status of malnourished HIV-infected patients but in terms of quality of nutritional improvement, subjects on spirulina were better off than subjects on soya beans. Secondly, nutritional rehabilitation improves on immune status with a consequent drop in viral load but further investigations on the antiviral effects of this alga and its clinical implications are strongly needed.

Azabji-Kenfack, M.; Dikosso, S. Edie; Loni, E.G.; Onana, E.A.; Sobngwi, E.; Gbaguidi, E.; Kana, A.L. Ngougni; Nguefack-Tsague, G.; Von der Weid, D.; Njoya, O.; Ngogang, J.