Brito, Anderson Fernandes de; Braconi, Carla Torres; Weidmann, Manfred; Dilcher, Meik; Alves, João Marcelo Pereira; Gruber, Arthur; Zanotto, Paolo Marinho de Andrade
The alphabaculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) is the world's most successful viral bioinsecticide. Through the 1980s and 1990s, this virus was extensively used for biological control of populations of Anticarsia gemmatalis (Velvetbean caterpillar) in soybean crops. During this period, genetic studies identified several variable loci in the AgMNPV; however, most of them were not characterized at the sequence level. In this study we report a full genome comparison among 17 wild-type isolates of AgMNPV. We found the pangenome of this virus to contain at least 167 hypothetical genes, 151 of which are shared by all genomes. The gene bro-a that might be involved in host specificity and carrying transporter is absent in some genomes, and new hypothetical genes were observed. Among these genes there is a unique rnf12-like gene, probably implicated in ubiquitination. Events of gene fission and fusion are common, as four genes have been observed as single or split open reading frames. Gains and losses of genomic fragments (from 20 to 900 bp) are observed within tandem repeats, such as in eight direct repeats and four homologous regions. Most AgMNPV genes present low nucleotide diversity, and variable genes are mainly located in a locus known to evolve through homologous recombination. The evolution of AgMNPV is mainly driven by small indels, substitutions, gain and loss of nucleotide stretches or entire coding sequences. These variations may cause relevant phenotypic alterations, which probably affect the infectivity of AgMNPV. This work provides novel information on genomic evolution of the AgMNPV in particular and of baculoviruses in general.
de Brito, Anderson Fernandes; Braconi, Carla Torres; Weidmann, Manfred; Dilcher, Meik; Alves, João Marcelo Pereira; Gruber, Arthur; Zanotto, Paolo Marinho de Andrade
The alphabaculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) is the world’s most successful viral bioinsecticide. Through the 1980s and 1990s, this virus was extensively used for biological control of populations of Anticarsia gemmatalis (Velvetbean caterpillar) in soybean crops. During this period, genetic studies identified several variable loci in the AgMNPV; however, most of them were not characterized at the sequence level. In this study we report a full genome comparison among 17 wild-type isolates of AgMNPV. We found the pangenome of this virus to contain at least 167 hypothetical genes, 151 of which are shared by all genomes. The gene bro-a that might be involved in host specificity and carrying transporter is absent in some genomes, and new hypothetical genes were observed. Among these genes there is a unique rnf12-like gene, probably implicated in ubiquitination. Events of gene fission and fusion are common, as four genes have been observed as single or split open reading frames. Gains and losses of genomic fragments (from 20 to 900 bp) are observed within tandem repeats, such as in eight direct repeats and four homologous regions. Most AgMNPV genes present low nucleotide diversity, and variable genes are mainly located in a locus known to evolve through homologous recombination. The evolution of AgMNPV is mainly driven by small indels, substitutions, gain and loss of nucleotide stretches or entire coding sequences. These variations may cause relevant phenotypic alterations, which probably affect the infectivity of AgMNPV. This work provides novel information on genomic evolution of the AgMNPV in particular and of baculoviruses in general. PMID:26615220
Braconi, Carla Torres; Ardisson-Araújo, Daniel Mendes Pereira; Paes Leme, Adriana Franco; Oliveira, Juliana Velasco de Castro; Pauletti, Bianca Alves; Garcia-Maruniak, Alejandra; Ribeiro, Bergmann Morais; Maruniak, James E; Zanotto, Paolo Marinho de Andrade
Baculoviruses infect insects, producing two distinct phenotypes during the viral life cycle: the budded virus (BV) and the occlusion-derived virus (ODV) for intra- and inter-host spread, respectively. Since the 1980s, several countries have been using Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) as a biological control agent against the velvet bean caterpillar, A. gemmatalis. The genome of AgMNPV isolate 2D (AgMNPV-2D) carries at least 152 potential genes, with 24 that possibly code for structural proteins. Proteomic studies have been carried out on a few baculoviruses, with six ODV and two BV proteomes completed so far. Moreover, there are limited data on virion proteins carried by AgMNPV-2D. Therefore, structural proteins of AgMNPV-2D were analysed by MALDI- quadrupole-TOF and liquid chromatography MS/MS. A total of 44 proteins were associated with the ODV and 33 with the BV of AgMNPV-2D. Although 38 structural proteins were already known, we found six new proteins in the ODV and seven new proteins carried by the AgMNPV-2D BV. Eleven cellular proteins that were found on several other enveloped viruses were also identified, which are possibly carried with the virion. These findings may provide novel insights into baculovirus biology and their host interaction. Moreover, our data may be helpful in subsequent applied studies aiming to improve AgMNPV use as a biopesticide and a biotechnology tool for gene expression or delivery.
de Rezende, Syomara Hakiko Matusita Soares; Castro, Maria Elita B; Souza, Marlinda L
Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) has been widely used to control the velvetbean caterpillar, Anticarsia gemmatalis, in Brazil. To date, AgMNPV has been produced by larval infection and, due to in vivo production limitations and the continuing high demand for the biopesticide, attempts should be made to develop in vitro production of this virus. In order to investigate the effects caused by serial passage of AgMNPV in cell culture, we carried out a total of ten passages and analyzed the morphological and the genomic changes of the virus. After six passages, the many-polyhedra (MP) phenotype started to switch to the few-polyhedra (FP) phenotype which rapidly accumulated in the virus population. Ultrastructural analysis showed typical signs of FP mutant formation such as decrease in the number of polyhedra per cell, polyhedra aberrant morphology and low numbers of virions occluded in the protein matrix. Also enhanced BV production was observed from the fifth passage indicating that FP mutants were becoming predominant in comparison to the wild type virus. Restriction endonuclease analysis of the viral DNA revealed that lower and higher passages had similar profiles indicating that there were no large insertions or deletions or rearrangements in their genomes and indicating the generation of FP mutants instead of defective interfering viruses.
Haase, Santiago; McCarthy, Christina B.; Ferrelli, M. Leticia; Pidre, Matias L.; Sciocco-Cap, Alicia; Romanowski, Victor
Anticarsia gemmatalis is an important pest in legume crops in South America and it has been successfully controlled using Anticarsia gemmatalis Multiple Nucleopolyhedrovirus (AgMNPV) in subtropical climate zones. Nevertheless, in temperate climates its speed of kill is too slow. Taking this into account, genetic modification of AgMNPV could lead to improvements of its biopesticidal properties. Here we report the generation of a two-component system that allows the production of recombinant AgMNPV. This system is based on a parental AgMNPV in which the polyhedrin gene (polh) was replaced by a bacterial β-galactosidase (lacZ) gene flanked by two target sites for the homing endonuclease I-PpoI. Co-transfection of insect cells with linearized (I-PpoI-digested) parental genome and a transfer vector allowed the restitution of polh and the expression of a heterologous gene upon homologous recombination, with a low background of non-recombinant AgMNPV. The system was validated by constructing a recombinant occlusion-positive (polh+) AgMNPV expressing the green fluorescent protein gene (gfp). This recombinant virus infected larvae normally per os and led to the expression of GFP in cell culture as well as in A. gemmatalis larvae. These results demonstrate that the system is an efficient method for the generation of recombinant AgMNPV expressing heterologous genes, which can be used for manifold purposes, including biotechnological and pharmaceutical applications and the production of orally infectious recombinants with improved biopesticidal properties. PMID:25835531
Pitta, Rafael M; Boiça, Arlindo L; Jesus, Flávio G de; Tagliari, Sônia R A
The velvetbean caterpillar Anticarsia gemmatalis Hübner attacks peanut leaves, and the use of resistant varieties has directly contributed to ecological and economic aspects of pest control. The aim of this work was to select resistant peanut genotypes to A. gemmatalis using cluster analyses (dendogram obtained by Ward's methods and K-means) and Principal Components analysis for data interpretation. The evaluated genotypes were: IAC 5, IAC 8112, IAC 22 and IAC Tatu ST with upright growth habit, and IAC 147, IAC 125, IAC Caiapó and IAC Runner 886 with runner growth habit, and soybean genotype BR 16 as a susceptible control. The biological parameters: leaf consumption, larval (4 masculine instar) and pupal (24h old) weight, larval and pupal development time and adult longevity were evaluated at laboratory conditions. The genotypes IAC 147 and IAC Runner 886 were resistant to A. gemmatalis in both cluster tests, grouping apart from most of the other genotypes. Both dendrogram and K-means methods provided satisfactory biological explanation, and they can be complementary used together with Principal Component and vice-versa. These results suggest that cluster analyses may be an important statistical tool in the selection of host plant resistance.
Silva, Farley W. S.; Viol, Daniel L.; Faria, Sirlene V.; Lima, Eraldo; Valicente, Fernando H.; Elliot, Simon L.
Defence from parasites and pathogens involves a cost. Thus, it is expected that organisms use this only at high population densities, where the risk of pathogen transmission may be high, as proposed by the "density-dependent prophylaxis" (DDP) hypothesis. These predictions have been tested in a wide range of insects, both in comparative and experimental studies. We think it pertinent to consider a continuum between solitarious and gregarious living insects, wherein: (1) solitarious insects are those that are constitutively solitary and do not express any phenotypic plasticity, (2) the middle of the continuum is represented by insects that are subject to fluctuations in local density and show a range of facultative and plastic changes; and (3) constitutively gregarious forms live gregariously and show the gregarious phenotype even in the absence of crowding stimuli. We aimed to chart some of the intermediary continuum with an insect that presents solitarious aspects, but that is subject to fluctuations in density. Thus, Anticarsia gemmatalis (Lepidoptera: Noctuidae) larvae reared at higher densities showed changes in coloration, a greater degree of encapsulation, had higher hemocyte densities and were more resistant to Baculovirus anticarsia, but not to Bacillus thuringiensis. Meanwhile, with increased rearing density there was reduced capsule melanization. Hemocyte density was the only variable that did not vary according to larval phenotype. The observed responses were not a continuous function of larval density, but an all-or-nothing response to the presence of a conspecific. As A. gemmatalis is not known for gregarious living, yet shows these density-dependent changes, it thus seems that this plastic phenotypic adjustment may be a broader phenomenon than previously thought. PMID:23626700
Biological characteristics of Anticarsia gemmatalis (Lepidoptera: Noctuidae) for three consecutive generations under different temperatures: understanding the possible impact of global warming on a soybean pest.
da Silva, D M; Hoffmann-Campo, C B; de Freitas Bueno, A; de Freitas Bueno, R C O; de Oliveira, M C N; Moscardi, F
Climate changes can affect the distribution and intensity of insect infestations through direct effects on their life cycles. Experiments were carried out during three consecutive generations to evaluate the effect of different temperatures (25°C, 28°C, 31°C, 34°C and 37±1°C) on biological traits of the velvetbean caterpillar Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Noctuidae). The insects were fed on artificial diet and reared in environmental chambers set at 14 h photophase. The developmental cycle slowed with the increase in the temperature, within the 25°C to 34°C range. Male and female longevities were reduced with an increase in temperature from 25°C to 28°C. Egg viability was highest at 25°C, and the sex ratio was not influenced by temperature, in the three generations. There was no interactive effect between development time and temperature on pupal weight. The results suggested that the increase in the temperature negatively impacted A. gemmatalis development inside the studied temperature range, indicating a possible future reduction of its occurrence on soybean crops, as a consequence of global warming, mainly considering its impact on tropical countries where this plant is cropped. A. gemmatalis was not able to adapt to higher temperatures in a three-generation interval for the studied temperature range. However, a gradual increase and a longer adaptation period may favor insect selection and consequently adaptation, and must be considered in future studies in this area. Moreover, it is important to consider that global warming might turn cold areas more suitable to A. gemmatalis outbreaks. Therefore, more than a future reduction of A. gemmatalis occurrence due to global warming, we might expect changes regarding its area of occurrence on a global perspective.
Lima, Anabele Azevedo; Aragão, Clara Wandenkolck Silva; de Castro, Maria Elita Batista; Oliveira, Juliana Velasco de Castro; Sosa Gómez, Daniel Ricardo; Ribeiro, Bergmann Morais
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
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.
Macedo, Maria L. R.; Kubo, Carlos E. G.; Freire, Maria G. M.; Júnior, Roberto T. A.; Parra, José R. P.
The effects of the beech apricot, Labramia bojeri A. de Candolle (Sapotales: Sapotaceae), seed aqueous extract on the larval development of the velvetbean moth, Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), was evaluated. The extract inhibited larval development, pupal weight, and survival and emergence of adults. Digestive proteolytic activity in larval midgut and feces extracts was determined. Larvae fed 10 g/L of the aqueous extract showed a significant reduction in trypsin activity (∼64%), when compared with control larvae. Trypsin and Chymotrypsin activities were also detected in fecal material in aqueous-extract-fed larvae, with about ∼4.5 times more trypsin activity than the controls. The results from dietary utilization experiments with A. gemmatalis larvae showed a reduction in the efficiency of conversion of ingested food and digested food and an increase in approximate digestibility and metabolic cost. The effect of the extract suggests the potential use of L. bojeri seeds to inhibit the development of A. gemmatalis via oral exposure. The L. bojeri extract can be an alternative to other methods of control. PMID:25373174
Macedo, Maria L R; Kubo, Carlos E G; Freire, Maria G M; Júnior, Roberto T A; Parra, José R P
The effects of the beech apricot, Labramia bojeri A. de Candolle (Sapotales: Sapotaceae), seed aqueous extract on the larval development of the velvetbean moth, Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), was evaluated. The extract inhibited larval development, pupal weight, and survival and emergence of adults. Digestive proteolytic activity in larval midgut and feces extracts was determined. Larvae fed 10 g/L of the aqueous extract showed a significant reduction in trypsin activity (~64%), when compared with control larvae. Trypsin and chymotrypsin activities were also detected in fecal material in aqueous-extract-fed larvae, with about ~4.5 times more trypsin activity than the controls. The results from dietary utilization experiments with A. gemmatalis larvae showed a reduction in the efficiency of conversion of ingested food and digested food and an increase in approximate digestibility and metabolic cost. The effect of the extract suggests the potential use of L. bojeri seeds to inhibit the development of A. gemmatalis via oral exposure. The L. bojeri extract can be an alternative to other methods of control.
Production of occlusion bodies of Anticarsia gemmatalis multiple nucleopolyhedrovirus in serum-free suspension cultures of the saUFL-AG-286 cell line: influence of infection conditions and statistical optimization.
Micheloud, Gabriela A; Gioria, Verónica V; Pérez, Gustavo; Claus, Juan D
The influence of the conditions of infection on the yield of occlusion bodies (OBs) of the Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV), produced in serum-free suspension cultures of saUFL-AG-286 cells, was investigated by two 2(2) full factorial experiments with centre point. Each experiment tested the effects of the initial cell density and the multiplicity of infection at two levels, in the four possible combinations of levels and conditions, plus a further combination with each condition set at the middle of its extreme levels. The yield of occlusion bodies proved to be sensitive to the modification of infection conditions. Maximum yield as high as 3 x 10(8) OBs mL(-1) was attained provided that the maximum density of viable cells was in the range between 4 and 8 x 10(5) cells mL(-1). The optimum value of the maximum density of viable cells could be reached by the combination of several values of initial cell density and multiplicity of infection. A regression model was established and validated in order to optimize the infection conditions. These results demonstrate the importance of an adequate selection of infection conditions, and they could be useful in the development of a feasible in vitro process to produce the AgMNPV insecticide in a new serum-free medium.
Soybean production is increasing around the world and, to no surprise, so are the reports of soybean diseases caused by Phytophthora sojae, including Phytophthora seed, root, and stem rot. Phytophthora sojae is a diploid oomycete, which is homothallic and is limited to primarily one host: the soybe...
Fang, Yufeng; Jang, Hyo Sang; Watson, Gregory W; Wellappili, Dulani P; Tyler, Brett M
To date, nuclear localization signals (NLSs) that target proteins to nuclei in oomycetes have not been defined, but have been assumed to be the same as in higher eukaryotes. Here, we use the soybean pathogen Phytophthora sojae as a model to investigate these sequences in oomycetes. By establishing a reliable in vivo NLS assay based on confocal microscopy, we found that many canonical monopartite and bipartite classical NLSs (cNLSs) mediated nuclear import poorly in P. sojae. We found that efficient localization of P. sojae nuclear proteins by cNLSs requires additional basic amino acids at distal sites or collaboration with other NLSs. We found that several representatives of another well-characterized NLS, proline-tyrosine NLS (PY-NLS) also functioned poorly in P. sojae. To characterize PY-NLSs in P. sojae, we experimentally defined the residues required by functional PY-NLSs in three P. sojae nuclear-localized proteins. These results showed that functional P. sojae PY-NLSs include an additional cluster of basic residues for efficient nuclear import. Finally, analysis of several highly conserved P. sojae nuclear proteins including ribosomal proteins and core histones revealed that these proteins exhibit a similar but stronger set of sequence requirements for nuclear targeting compared with their orthologs in mammals or yeast.
Fang, Yufeng; Jang, Hyo Sang; Watson, Gregory W.; Wellappili, Dulani P.; Tyler, Brett M.
To date, nuclear localization signals (NLSs) that target proteins to nuclei in oomycetes have not been defined, but have been assumed to be the same as in higher eukaryotes. Here, we use the soybean pathogen Phytophthora sojae as a model to investigate these sequences in oomycetes. By establishing a reliable in vivo NLS assay based on confocal microscopy, we found that many canonical monopartite and bipartite classical NLSs (cNLSs) mediated nuclear import poorly in P. sojae. We found that efficient localization of P. sojae nuclear proteins by cNLSs requires additional basic amino acids at distal sites or collaboration with other NLSs. We found that several representatives of another well-characterized NLS, proline-tyrosine NLS (PY-NLS) also functioned poorly in P. sojae. To characterize PY-NLSs in P. sojae, we experimentally defined the residues required by functional PY-NLSs in three P. sojae nuclear-localized proteins. These results showed that functional P. sojae PY-NLSs include an additional cluster of basic residues for efficient nuclear import. Finally, analysis of several highly conserved P. sojae nuclear proteins including ribosomal proteins and core histones revealed that these proteins exhibit a similar but stronger set of sequence requirements for nuclear targeting compared with their orthologs in mammals or yeast. PMID:28210240
Shrestha, Sirjana Devi; Chapman, Patrick; Zhang, Yun; Gijzen, Mark
The Phytophthora sojae avirulence gene Avr3a encodes an effector that is capable of triggering immunity on soybean plants carrying the resistance gene Rps3a. P. sojae strains that express Avr3a are avirulent to Rps3a plants, while strains that do not are virulent. To study the inheritance of Avr3a expression and virulence towards Rps3a, genetic crosses and self-fertilizations were performed. A cross between P. sojae strains ACR10 X P7076 causes transgenerational gene silencing of Avr3a allele, and this effect is meiotically stable up to the F5 generation. However, test-crosses of F1 progeny (ACR10 X P7076) with strain P6497 result in the release of silencing of Avr3a. Expression of Avr3a in the progeny is variable and correlates with the phenotypic penetrance of the avirulence trait. The F1 progeny from a direct cross of P6497 X ACR10 segregate for inheritance for Avr3a expression, a result that could not be explained by parental imprinting or heterozygosity. Analysis of small RNA arising from the Avr3a gene sequence in the parental strains and hybrid progeny suggests that the presence of small RNA is necessary but not sufficient for gene silencing. Overall, we conclude that inheritance of the Avr3a gene silenced phenotype relies on factors that are variable among P. sojae strains. PMID:26930612
A number of moth species responded positively to phenylacetaldehyde (PAA) and to the binary blend of PAA + linalool in tests conducted in peanut fields in northern Florida, USA. Velvetbean caterpillar moths (Anticarsia gemmatalis Hübner) were the most commonly collected species, with almost 13,000 ...
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...
Evaluation of combinations of flower odor compounds in northern Florida, revealed that linalool was synergistic in attractiveness with phenylacetaldehyde (PAA) to the migratory moth velvetbean caterpillar (Anticarsia gemmatalis Hübner). This noctuid was the most common species collected from traps w...
A neem formulation (Neemix® 4.5) and spinosad (SpinTor® 2SC) were tested for their effects when mixed with the multicapsid nucleopolyhedrovirus virus (AgMNPV) from the velvetbean caterpillar, Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), for control of pickleworm larvae, Diaphania nitidalis...
Phytophthora sojae (Kaufm. & Gerd.) causes seed rot, pre- and post-emergence damping off, and sometimes foliar blight in soybean (Glycine max). Crop loss may approach 100% with susceptible cultivars. We report here the development of a unique quantitative PCR assay specific to DNA of P. sojae, and a...
Li, Delong; Lv, Binna; Tan, Lingling; Yang, Qianqian; Liang, Wenxing
Lysine acetylation is a dynamic and highly conserved post-translational modification that plays an important regulatory role in almost every aspects of cell metabolism in both eukaryotes and prokaryotes. Phytophthora sojae is one of the most important plant pathogens due to its huge economic impact. However, to date, little is known about the functions of lysine acetylation in this Phytopthora. Here, we conducted a lysine acetylome in P. sojae. Overall, 2197 lysine acetylation sites in 1150 proteins were identified. The modified proteins are involved in diverse biological processes and are localized to multiple cellular compartments. Importantly, 7 proteins involved in the pathogenicity or the secretion pathway of P. sojae were found to be acetylated. These data provide the first comprehensive view of the acetylome of P. sojae and serve as an important resource for functional analysis of lysine acetylation in plant pathogens.
Li, Delong; Lv, Binna; Tan, Lingling; Yang, Qianqian; Liang, Wenxing
Lysine acetylation is a dynamic and highly conserved post-translational modification that plays an important regulatory role in almost every aspects of cell metabolism in both eukaryotes and prokaryotes. Phytophthora sojae is one of the most important plant pathogens due to its huge economic impact. However, to date, little is known about the functions of lysine acetylation in this Phytopthora. Here, we conducted a lysine acetylome in P. sojae. Overall, 2197 lysine acetylation sites in 1150 proteins were identified. The modified proteins are involved in diverse biological processes and are localized to multiple cellular compartments. Importantly, 7 proteins involved in the pathogenicity or the secretion pathway of P. sojae were found to be acetylated. These data provide the first comprehensive view of the acetylome of P. sojae and serve as an important resource for functional analysis of lysine acetylation in plant pathogens. PMID:27412925
Martin, Frank N.; Douda, Bensasson; Tyler, Brett M.; Boore,Jeffrey L.
The complete sequences of the mitochondrial genomes of theoomycetes of Phytophthora ramorum and P. sojae were determined during thecourse of their complete nuclear genome sequencing (Tyler, et al. 2006).Both are circular, with sizes of 39,314 bp for P. ramorum and 42,975 bpfor P. sojae. Each contains a total of 37 identifiable protein-encodinggenes, 25 or 26 tRNAs (P. sojae and P. ramorum, respectively)specifying19 amino acids, and a variable number of ORFs (7 for P. ramorum and 12for P. sojae) which are potentially additional functional genes.Non-coding regions comprise approximately 11.5 percent and 18.4 percentof the genomes of P. ramorum and P. sojae, respectively. Relative to P.sojae, there is an inverted repeat of 1,150 bp in P. ramorum thatincludes an unassigned unique ORF, a tRNA gene, and adjacent non-codingsequences, but otherwise the gene order in both species is identical.Comparisons of these genomes with published sequences of the P. infestansmitochondrial genome reveals a number of similarities, but the gene orderin P. infestans differs in two adjacent locations due to inversions.Sequence alignments of the three genomes indicated sequence conservationranging from 75 to 85 percent and that specific regions were morevariable than others.
Wong, James; Gao, Lei; Yang, Yang; Zhai, Jixian; Arikit, Siwaret; Yu, Yu; Duan, Shuyi; Chan, Vicky; Xiong, Qin; Yan, Jun; Li, Shengben; Liu, Renyi; Wang, Yuanchao; Tang, Guiliang; Meyers, Blake C; Chen, Xuemei; Ma, Wenbo
The genus Phytophthora consists of many notorious pathogens of crops and forestry trees. At present, battling Phytophthora diseases is challenging due to a lack of understanding of their pathogenesis. We investigated the role of small RNAs in regulating soybean defense in response to infection by Phytophthora sojae, the second most destructive pathogen of soybean. Small RNAs, including microRNAs (miRNAs) and small interfering RNAs (siRNAs), are universal regulators that repress target gene expression in eukaryotes. We identified known and novel small RNAs that differentially accumulated during P. sojae infection in soybean roots. Among them, miR393 and miR166 were induced by heat-inactivated P. sojae hyphae, indicating that they may be involved in soybean basal defense. Indeed, knocking down the level of mature miR393 led to enhanced susceptibility of soybean to P. sojae; furthermore, the expression of isoflavonoid biosynthetic genes was drastically reduced in miR393 knockdown roots. These data suggest that miR393 promotes soybean defense against P. sojae. In addition to miRNAs, P. sojae infection also resulted in increased accumulation of phased siRNAs (phasiRNAs) that are predominantly generated from canonical resistance genes encoding nucleotide binding-leucine rich repeat proteins and genes encoding pentatricopeptide repeat-containing proteins. This work identifies specific miRNAs and phasiRNAs that regulate defense-associated genes in soybean during Phytophthora infection.
Phytophthora root and stem rot is one of the most yield-limiting diseases of soybean [Glycine max (L.) Merr], caused by the oomycete Phytophthora sojae. Partial resistance is controlled by several genes and, compared to single gene (Rps gene) resistance to P. sojae, places less selection pressure on...
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...
Janknegt, Robert; Scott, Mike; Mairs, Jill; Timoney, Mark; McElnay, James; Brenninkmeijer, Rob
Drug selection should be a rational process that embraces the principles of evidence-based medicine. However, many factors may affect the choice of agent. It is against this background that the System of Objectified Judgement Analysis (SOJA) process for rational drug-selection was developed. This article describes how the information on which the SOJA process is based, was researched and processed.
Kang, Heonil; Eun, Geun; Ha, Jihye; Kim, Yongchul; Park, Namsook; Kim, Donggeun; Choi, Insoo
A new soybean cyst nematode Heterodera sojae n. sp. was found from the roots of soybean plants in Korea. Cysts of H. sojae n. sp. appeared more round, shining, and darker than that of H. glycines. Morphologically, H. sojae n. sp. differed from H. glycines by fenestra length (23.5–54.2 µm vs. 30–70 µm), vulval silt length (9.0–24.4 µm vs. 43–60 µm), tail length of J2 (54.3–74.8 µm vs. 40–61 µm), and hyaline part of J2 (32.6–46.3 µm vs. 20–30 µm). It is distinguished from H. elachista by larger cyst (513.4–778.3 µm × 343.4–567.1 µm vs. 350–560 µm × 250–450 µm) and longer stylet length of J2 (23.8–25.3 µm vs. 17–19 µm). Molecular analysis of rRNA large subunit (LSU) D2–D3 segments and ITS gene sequence shows that H. sojae n. sp. is more close to rice cyst nematode H. elachista than H. glycines. Heterodera sojae n. sp. was widely distributed in Korea. It was found from soybean fields of all three provinces sampled. PMID:28154434
Chen, Rui; Hu, Zheng; Zhang, Hui
MicroRNAs (miRNAs) play important roles in post-transcriptional gene silencing by directing target mRNA cleavage or translational inhibition. Currently, hundreds of miRNAs have been identified in plants, but no report has been published of wild soybean (Glycine soja Sieb). We constructed a small-RNA library consisting of 2 880 sequences with high quality, in which 1 347 were 19-24 nt in length. By utilizing the miRNA, Rfam and domesticated soybean expressed sequence tag database, we have analyzed and predicted the secondary structure of these small RNAs. As a result, 15 conserved miRNA candidates belonging to eight different families and nine novel miRNA candidates comprising eight families were identified in wild soybean seedlings. All these miRNA candidates were validated by northern blot and the novel candidates expressed in a tissue-specific manner. Furthermore, putative target genes were predicted for novel miRNA candidates and two of them were verified by 5'-rapid amplification of cDNA ends experiments. These results provided useful information for miRNA research in wild soybean and plants.
Fronza, Edegar; Migues, Ignacio; Specht, Alexandre; de Barros, Neiva Monteiro; Heinzen, Horacio
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, α-tocopherol and α-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.
Phytophthora encodes an unusually large number of glycosyl hydrolases (GH), with many large gene families resulting from duplication events. There are ten copies of GH 12 (cel12) present in Phytophthora sojae. This is the only pathogen endoglucanase family to which plants produce an inhibitory pr...
Zhou, Yan-Ying; Luo, Shi-Hong; Yi, Ting-Shuang; Li, Chun-Huan; Luo, Qian; Hua, Juan; Liu, Yan; Li, Sheng-Hong
The wild soybean (Glycine soja Sieb. et Zucc) has been reported to be relatively resistant to insect and pathogenic pests. However, the responsible secondary metabolites in the aerial part of this important plant are largely unknown. From the aerial part of G. soja, 13 compounds were isolated and identified, including seven isoflavonoids (1-7), a cyclitol (8), two sterol derivatives (9 and 10), and three triterpenoids (11-13). Compound 7 is a new isoflavonoid, and compounds 9 and 10 are reported as natural products for the first time. The growth inhibitory activity of 1, 3, 4, and 8 against the larvae of Spodoptera litura was investigated. The most abundant isoflavonoid in the aerial part of G. soja, daidzein (1), which could not be metabolized by S. litura, was found to inhibit the insect larvae growth significantly in 3 days after feeding diets containing the compound. Compounds 3, 4, and 8, which could be partially or completely metabolized, were inactive. Our results suggested that the isoflavonoid daidzein (1) might function as a constitutive defense component in G. soja against insect pests.
Phytophthora stem and root rot disease, caused by Phytophthora sojae, is one of the most destructive diseases of soybean (Glycine max (L.) Merr.), and has been increasing in several soybean-producing areas around the world. This disease induces serious limitations on soybean production, with yield l...
Phytophthora root and stem rot caused by Phytophthora sojae Kaufmann and Gerdmann is one of the most severe soybean [Glycine max (L.) Merr] diseases in the US. Partial resistance is as effective in managing this disease as single-gene (Rps) mediated resistance and is more durable. The objective of t...
Phytophthora sojae causes damping off, root rot, and stem rot of soybean, particularly in poorly drained soils. The use of resistance has been one of the primary management tools used to control this disease, with the most commonly used genes being Rps1c and Rps1k, followed by Rps1a. The Varietal In...
Aspergillus section Flavi includes aflatoxin-producing and nonproducing fungi. A. sojae is unable to produce aflatoxins and is generally recognized as safe for food fermentation. However, because of its taxonomical relatedness to aflatoxin-producing A. parasiticus and A. flavus, it is necessary to...
Ye, Wenwu; Wang, Yang; Tyler, Brett M.; Wang, Yuanchao
Comparative genomic analysis is useful for identifying genes affected by evolutionary selection and for studying adaptive variation in gene functions. In Phytophthora sojae, a model oomycete plant pathogen, the related study is lacking. We compared sequence data among four isolates of P. sojae, which represent its four major genotypes. These isolates exhibited >99.688%, >99.864%, and >98.981% sequence identities at genome, gene, and non-gene regions, respectively. One hundred and fifty-three positive selection and 139 negative selection candidate genes were identified. Between the two categories of genes, the positive selection genes were flanked by larger intergenic regions, poorly annotated in function, and less conserved; they had relatively lower transcription levels but many genes had increased transcripts during infection. Genes coding for predicted secreted proteins, particularly effectors, were overrepresented in positive selection. Several RxLR effector genes were identified as positive selection genes, exhibiting much stronger positive selection levels. In addition, candidate genes with presence/absence polymorphism were analyzed. This study provides a landscape of genomic variation among four representative P. sojae isolates and characterized several evolutionary selection-affected gene candidates. The results suggest a relatively covert two-speed genome evolution pattern in P. sojae and will provide clues for identification of new virulence factors in the oomycete plant pathogens. PMID:27746768
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.
Wang, Ke-Jing; Li, Xiang-Hua; Liu, Yang
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.
Aflatoxins are the most toxic and carcinogenic secondary metabolites produced primarily by the filamentous fungi Aspergillus flavus and Aspergillus parasiticus. The toxins cause devastating economic losses because of strict regulations on distribution of contaminated products. Aspergillus sojae are...
Zhou, San; Sekizaki, Haruo; Yang, Zhihong; Sawa, Satoko; Pan, Jun
Hardseededness in annual wild soybean (Glycine soja Sieb. Et Zucc.) is a valuable trait that affects the germination, viability, and quality of stored seeds. Two G. soja ecotypes native to Shandong Province of China have been used to identify the phenolics in the seed coat that correlate with the seed hardness and seed germination. Three major phenolics from the seed coat were isolated and identified as epicatechin, cyanidin 3-O-glucoside, and delphinidin 3-O-glucoside. Of the three phenolics, only the change of epicatechin exhibited a significant positive correlation with the change of hard seed percentages both under different water conditions during seed development and under different gas conditions during seed storage. Epicatechin also reveals a hormesis-like effect on the seed germination of G. soja. Epicatechin is suggested to be functionally related to coat-imposed hardseededness in G. soja.
Mora-Lugo, Rodrigo; Madrigal, Marvin; Yelemane, Vikas; Fernandez-Lahore, Marcelo
The biotechnological value of Aspergillus sojae ATCC 20235 (A. sojae) for production of pectinases in solid-state fermentation (SSF) has been demonstrated recently. However, a common drawback of fungal solid-state cultures is the poor diffusion of oxygen into the fungi that limits its growth and biological productivity. The bacterial Vitreoscilla hemoglobin (VHb) has favored the metabolism and productivities of various bacterial and yeast strains besides alleviating hypoxic conditions of its native host, but the use of VHb in filamentous fungi still remains poor explored. Based on the known effects of VHb, this study assessed its applicability to improve A. sojae performance in SSF. The VHb gene (vgb) under control of the constitutive Aspergillus nidulants gpdA promoter was introduced into the genome of A. sojae by Agrobacterium-mediated transformation. Successful fungal transformants were identified by fluorescence microscopy and polymerase chain reaction (PCR) analyses. In solid-state cultures, the content of protease, exo-polygalacturonase (exo-PG), and exo-polymethylgalacturonase (exo-PMG) of the transformed fungus (A. sojae vgb+) improved were 26, 60, and 44 % higher, respectively, in comparison to its parental strain (A. sojae wt). Similarly, biomass content was also 1.3 times higher in the transformant strain. No significant difference was observed in endo-polygalacturonase (endo-PG) content between both fungal strains, suggesting dissimilar effects of VHb towards different enzymatic productions. Overall, our results show that biomass, protease, and exo-pectinase content of A. sojae in SSF can be improved by transformation with VHb.
Kim, Moon Young; Lee, Sunghoon; Van, Kyujung; Kim, Tae-Hyung; Jeong, Soon-Chun; Choi, Ik-Young; Kim, Dae-Soo; Lee, Yong-Seok; Park, Daeui; Ma, Jianxin; Kim, Woo-Yeon; Kim, Byoung-Chul; Park, Sungjin; Lee, Kyung-A; Kim, Dong Hyun; Kim, Kil Hyun; Shin, Jin Hee; Jang, Young Eun; Kim, Kyung Do; Liu, Wei Xian; Chaisan, Tanapon; Kang, Yang Jae; Lee, Yeong-Ho; Kim, Kook-Hyung; Moon, Jung-Kyung; Schmutz, Jeremy; Jackson, Scott A; Bhak, Jong; Lee, Suk-Ha
The genome of soybean (Glycine max), a commercially important crop, has recently been sequenced and is one of six crop species to have been sequenced. Here we report the genome sequence of G. soja, the undomesticated ancestor of G. max (in particular, G. soja var. IT182932). The 48.8-Gb Illumina Genome Analyzer (Illumina-GA) short DNA reads were aligned to the G. max reference genome and a consensus was determined for G. soja. This consensus sequence spanned 915.4 Mb, representing a coverage of 97.65% of the G. max published genome sequence and an average mapping depth of 43-fold. The nucleotide sequence of the G. soja genome, which contains 2.5 Mb of substituted bases and 406 kb of small insertions/deletions relative to G. max, is ∼0.31% different from that of G. max. In addition to the mapped 915.4-Mb consensus sequence, 32.4 Mb of large deletions and 8.3 Mb of novel sequence contigs in the G. soja genome were also detected. Nucleotide variants of G. soja versus G. max confirmed by Roche Genome Sequencer FLX sequencing showed a 99.99% concordance in single-nucleotide polymorphism and a 98.82% agreement in insertion/deletion calls on Illumina-GA reads. Data presented in this study suggest that the G. soja/G. max complex may be at least 0.27 million y old, appearing before the relatively recent event of domestication (6,000∼9,000 y ago). This suggests that soybean domestication is complicated and that more in-depth study of population genetics is needed. In any case, genome comparison of domesticated and undomesticated forms of soybean can facilitate its improvement.
Cheng, Qun; Li, Ninghui; Dong, Lidong; Zhang, Dayong; Fan, Sujie; Jiang, Liangyu; Wang, Xin; Xu, Pengfei; Zhang, Shuzhen
Isoflavone reductase (IFR) is an enzyme involved in the biosynthetic pathway of isoflavonoid phytoalexin in plants. IFRs are unique to the plant kingdom and are considered to have crucial roles in plant response to various biotic and abiotic environmental stresses. Here, we report the characterization of a novel member of the soybean isoflavone reductase gene family GmIFR. Overexpression of GmIFR transgenic soybean exhibited enhanced resistance to Phytophthora sojae. Following stress treatments, GmIFR was significantly induced by P. sojae, ethephon (ET), abscisic acid (placeCityABA), salicylic acid (SA). It is located in the cytoplasm when transiently expressed in soybean protoplasts. The daidzein levels reduced greatly for the seeds of transgenic plants, while the relative content of glyceollins in transgenic plants was significantly higher than that of non-transgenic plants. Furthermore, we found that the relative expression levels of reactive oxygen species (ROS) of transgenic soybean plants were significantly lower than those of non-transgenic plants after incubation with P. sojae, suggesting an important role of GmIFR might function as an antioxidant to reduce ROS in soybean. The enzyme activity assay suggested that GmIFR has isoflavone reductase activity. PMID:26635848
Cheng, Qun; Li, Ninghui; Dong, Lidong; Zhang, Dayong; Fan, Sujie; Jiang, Liangyu; Wang, Xin; Xu, Pengfei; Zhang, Shuzhen
Isoflavone reductase (IFR) is an enzyme involved in the biosynthetic pathway of isoflavonoid phytoalexin in plants. IFRs are unique to the plant kingdom and are considered to have crucial roles in plant response to various biotic and abiotic environmental stresses. Here, we report the characterization of a novel member of the soybean isoflavone reductase gene family GmIFR. Overexpression of GmIFR transgenic soybean exhibited enhanced resistance to Phytophthora sojae. Following stress treatments, GmIFR was significantly induced by P. sojae, ethephon (ET), abscisic acid (placeCityABA), salicylic acid (SA). It is located in the cytoplasm when transiently expressed in soybean protoplasts. The daidzein levels reduced greatly for the seeds of transgenic plants, while the relative content of glyceollins in transgenic plants was significantly higher than that of non-transgenic plants. Furthermore, we found that the relative expression levels of reactive oxygen species (ROS) of transgenic soybean plants were significantly lower than those of non-transgenic plants after incubation with P. sojae, suggesting an important role of GmIFR might function as an antioxidant to reduce ROS in soybean. The enzyme activity assay suggested that GmIFR has isoflavone reductase activity.
Powell, W.; Morgante, M.; Doyle, J. J.; McNicol, J. W.; Tingey, S. V.; Rafalski, A. J.
A combination of nuclear and chloroplast simple sequence repeats (SSRs) have been used to investigate the levels and pattern of variability detected in Glycine max and G. soja genotypes. Based on the analysis of 700 soybean genotypes with 115 restriction fragment length polymorphism (RFLP) probes, 12 accessions were identified that represent 92% of the allelic variability detected in this genepool. These 12 core genotypes together with a sample of G. max and G. soja accessions were evaluated with 11 nuclear SSRs that detected 129 alleles. Compared with the other G. max and G. soja genotypes sampled, the core genotypes represent 40% of the allelic variability detected with SSRs. Despite the multi-allelic nature of soybean SSRs, dendrograms representing phenetic relationships between accessions clustered according to their subspecies origin. In addition to biparentally inherited nuclear SSRs, two uniparentally (maternally) transmitted chloroplast SSRs were also studied. A total of seven haplotypes were identified, and diversity indices of 0.405 +/- 0.088 and 0.159 +/- 0.071 were obtained for the two chloroplast SSRs. The availability of polymorphic SSR loci in the chloroplast genome provides new opportunities to investigate cytonuclear interactions in plants. PMID:8889540
Phytophthora root rot of soybean (Glycine max Merr.) is caused by the oomycete Phytophthora sojae (Kaufm. and Gerd.). P. sojae has a narrow host range, consisting primarily of soybean, and it is a serious pathogen worldwide. It exists in root and stem tissues as mycelium, wherein it can form oospo...
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...
Sahoo, Dipak K.; Abeysekara, Nilwala S.; Cianzio, Silvia R.; Robertson, Alison E.
Phytophthora sojae Kaufmann and Gerdemann, which causes Phytophthora root rot, is a widespread pathogen that limits soybean production worldwide. Development of Phytophthora resistant cultivars carrying Phytophthora resistance Rps genes is a cost-effective approach in controlling this disease. For this mapping study of a novel Rps gene, 290 recombinant inbred lines (RILs) (F7 families) were developed by crossing the P. sojae resistant cultivar PI399036 with the P. sojae susceptible AR2 line, and were phenotyped for responses to a mixture of three P. sojae isolates that overcome most of the known Rps genes. Of these 290 RILs, 130 were homozygous resistant, 12 heterzygous and segregating for Phytophthora resistance, and 148 were recessive homozygous and susceptible. From this population, 59 RILs homozygous for Phytophthora sojae resistance and 61 susceptible to a mixture of P. sojae isolates R17 and Val12-11 or P7074 that overcome resistance encoded by known Rps genes mapped to Chromosome 18 were selected for mapping novel Rps gene. A single gene accounted for the 1:1 segregation of resistance and susceptibility among the RILs. The gene encoding the Phytophthora resistance mapped to a 5.8 cM interval between the SSR markers BARCSOYSSR_18_1840 and Sat_064 located in the lower arm of Chromosome 18. The gene is mapped 2.2 cM proximal to the NBSRps4/6-like sequence that was reported to co-segregate with the Phytophthora resistance genes Rps4 and Rps6. The gene is mapped to a highly recombinogenic, gene-rich genomic region carrying several nucleotide binding site-leucine rich repeat (NBS-LRR)-like genes. We named this novel gene as Rps12, which is expected to be an invaluable resource in breeding soybeans for Phytophthora resistance. PMID:28081566
Zhang, Meixiang; Li, Qi; Liu, Tingli; Liu, Li; Shen, Danyu; Zhu, Ye; Liu, Peihan; Zhou, Jian-Min; Dou, Daolong
Plant pathogenic oomycetes, such as Phytophthora sojae, secrete an arsenal of host cytoplasmic effectors to promote infection. We have shown previously that P. sojae PsCRN63 (for crinkling- and necrosis-inducing proteins) induces programmed cell death (PCD) while PsCRN115 blocks PCD in planta; however, they are jointly required for full pathogenesis. Here, we find that PsCRN63 alone or PsCRN63 and PsCRN115 together might suppress the immune responses of Nicotiana benthamiana and demonstrate that these two cytoplasmic effectors interact with catalases from N. benthamiana and soybean (Glycine max). Transient expression of PsCRN63 increases hydrogen peroxide (H2O2) accumulation, whereas PsCRN115 suppresses this process. Transient overexpression of NbCAT1 (for N. benthamiana CATALASE1) or GmCAT1 specifically alleviates PsCRN63-induced PCD. Suppression of the PsCRN63-induced PCD by PsCRN115 is compromised when catalases are silenced in N. benthamiana. Interestingly, the NbCAT1 is recruited into the plant nucleus in the presence of PsCRN63 or PsCRN115; NbCAT1 and GmCAT1 are destabilized when PsCRN63 is coexpressed, and PsCRN115 inhibits the processes. Thus, PsCRN63/115 manipulates plant PCD through interfering with catalases and perturbing H2O2 homeostasis. Furthermore, silencing of catalase genes enhances susceptibility to Phytophthora capsici, indicating that catalases are essential for plant resistance. Taken together, we suggest that P. sojae secretes these two effectors to regulate plant PCD and H2O2 homeostasis through direct interaction with catalases and, therefore, overcome host immune responses. PMID:25424308
Zhao, Liang; Fan, Miaochun; Zhang, Dehui; Yang, Ruiping; Zhang, Feilong; Xu, Lin; Wei, Xiuli; Shen, Yaoyao; Wei, Gehong
A total of 155 nodule isolates that originated from seven sites in Northwest China were characterized by PCR-RFLP of the 16S rRNA gene and sequence analysis of multiple core genes (16S rRNA, recA, atpD, and glnII) in order to investigate the diversity and biogeography of Glycine soja-nodulating rhizobia. Among the isolates, 80 were Ensifer fredii, 19 were Ensifer morelense, 49 were Rhizobium radiobacter, and 7 were putative novel Rhizobium species. The phylogenies of E. fredii and E. morelense isolates in a concatenate tree (assembly of all housekeeping genes) were generally consistent with those in individual gene trees. However, incongruence was found in the phylogenies of the different genes of Rhizobium isolates, indicating that lateral transfer or recombination possibly occurred in these gene loci. Despite their species identity, all the isolates in this study formed a single lineage related to E. fredii in nodAand nifH gene phylogenies, which also indicated that the symbiotic genes were laterally transferred between different species. Biogeographic patterns were found at the species and strain genomic type levels, as revealed by BOXA1R fingerprinting, demonstrating that the evolution of rhizobial populations in different geographic locations was related to soil types, altitude and spatial effects. This study is the first to report that E. morelense, R. radiobacter, and Rhizobium sp. are microsymbionts of G. soja, as well as showing that the diversity of G. soja rhizobia is enhanced and new rhizobia have evolved in Northwest China.
Winter, Shawn M J; Shelp, Barry J; Anderson, Terry R; Welacky, Tom W; Rajcan, Istvan
Soybean cyst nematode (Heterodera glycines Ichinohe; SCN) is the primary disease responsible for yield loss of soybean [Glycine max (L.) Merr.]. Resistant cultivars are an effective management tool; however, the sources currently available have common resistant genes. Glycine soja Sieb. and Zucc., the wild ancestor of domesticated soybean, represents a diverse germplasm pool with known SCN resistance. The objectives of this research were to: (1) determine the genetic variation and inheritance of SCN resistance in a G. max ('S08-80') x G. soja (PI464925B) F (4:5) recombinant inbred line (RIL) population; and (2) identify and evaluate quantitative trait loci (QTL) associated with SCN resistance. Transgressive segregation for resistance was observed, although neither parent was resistant to the Chatham and Ruthven SCN isolates. Broad sense heritability was 0.81 for the Ruthven and 0.91 for the Chatham isolate. Root dry weight was a significant covariate that influenced cyst counts. One RIL [female index (FI) = 5.2 +/- 1.11] was identified as resistant to the Chatham isolate (FI < 10). Seventeen and three RILs infected with Chatham and Ruthven isolates, respectively, had mean adjusted cyst counts of zero. Unique and novel QTL, which derived resistance from G. soja, were identified on linkage groups I, K, and O, and individually explained 8, 7 and 5% (LOD = 2.1-2.7) of the total phenotypic variation, respectively. Significant epistatic interactions were found between pairs of SSR markers that individually may or may not have been associated with SCN resistance, which explained between 10 and 15% of the total phenotypic variation. Best-fit regression models explained 21 and 31% of the total phenotypic variation in the RIL population to the Chatham and Ruthven isolates, respectively. The results of this study help to improve the understanding of the genetic control of SCN resistance in soybean caused by minor genes resulting in horizontal resistance. The incorporation of
Zhao, Yuanling; Chang, Xin; Qi, Dongyue; Dong, Lidong; Wang, Guangjin; Fan, Sujie; Jiang, Liangyu; Cheng, Qun; Chen, Xi; Han, Dan; Xu, Pengfei; Zhang, Shuzhen
Phytophthora root and stem rot of soybean caused by the oomycete Phytophthora sojae, is a destructive disease worldwide. Ethylene response factors (ERFs) play important roles in regulating plant biotic and abiotic stress tolerance. In this study, a new ERF gene, GmERF113, was isolated from the highly resistant soybean ‘Suinong 10.’ Sequence analysis suggested that the protein encoded by GmERF113 contained a conserved AP2/ERF domain of 58 amino acid and belonged to the B-4 subgroup of the ERF subfamily. Expression of GmERF113 was significantly induced by P. sojae, ethylene, and methyl jasmonate. GmERF113 protein localized to the nucleus when transiently expressed in Arabidopsis protoplasts, could bind to the GCC-box, and acted as a transcription activator. In addition, a region of the full-length GmERF113, GmERF113-II, interacted with a basic helix-loop-helix transcription factor (GmbHLH) in yeast cells. Full-length GmERF113 also interacted with GmbHLH in planta. GmERF113-overexpressing transgenic plants in susceptible cultivar ‘Dongnong 50’ soybean exhibited increased resistance to P. sojae and positively regulated the expression of the pathogenesis-related genes, PR1 and PR10-1. These results indicate that GmERF113 may play a crucial role in the defense of soybean against P. sojae infection. PMID:28326092
Partial resistance to Phytophthora sojae in soybean is controlled by multiple quantitative trait loci (QTL). With traditional QTL mapping approaches, power to detect these QTL, frequently of small effect, can be limited by population size. Joint linkage QTL analysis of nested recombinant inbred li...
Phytophthora root and stem rot (PRR) of soybean, caused by Phytophthora sojae, is effectively controlled by Rps genes in soybean. Rps genes are race-specific, yet the mechanism of resistance, as well as susceptibility, remains largely unclear. Taking advantage of RNA-seq technology, we sequenced the...
Phytophthora root rot (PRR) caused by Phytophthora sojae Kaufm. & Gerd. and flooding can limit growth and productivity, of soybean [Glycine max (L.) Merr.], especially on poorly drained soils. The primary objective of this research project was to map quantitative trait loci (QTL) associated with f...
Qiu, Jie; Wang, Yu; Wu, Sanling; Wang, Ying-Ying; Ye, Chu-Yu; Bai, Xuefei; Li, Zefeng; Yan, Chenghai; Wang, Weidi; Wang, Ziqiang; Shu, Qingyao; Xie, Jiahua; Lee, Suk-Ha; Fan, Longjiang
Semi-wild soybean is a unique type of soybean that retains both wild and domesticated characteristics, which provides an important intermediate type for understanding the evolution of the subgenus Soja population in the Glycine genus. In this study, a semi-wild soybean line (Maliaodou) and a wild line (Lanxi 1) collected from the lower Yangtze regions were deeply sequenced while nine other semi-wild lines were sequenced to a 3-fold genome coverage. Sequence analysis revealed that (1) no independent phylogenetic branch covering all 10 semi-wild lines was observed in the Soja phylogenetic tree; (2) besides two distinct subpopulations of wild and cultivated soybean in the Soja population structure, all semi-wild lines were mixed with some wild lines into a subpopulation rather than an independent one or an intermediate transition type of soybean domestication; (3) high heterozygous rates (0.19-0.49) were observed in several semi-wild lines; and (4) over 100 putative selective regions were identified by selective sweep analysis, including those related to the development of seed size. Our results suggested a hybridization origin for the semi-wild soybean, which makes a complex Soja population structure.
Fang, Yufeng; Cui, Linkai; Gu, Biao; Arredondo, Felipe; Tyler, Brett M
Phytophthora is a filamentous fungus-like microorganism, but belongs to the oomycetes, in the kingdom Stramenopila. Phytophthora species are notorious as plant destroyers, causing multibillion-dollar damage to agriculture and natural ecosystems worldwide annually. For a long time, genome editing has been unattainable in oomycetes, because of their extremely low rate of homologous recombination. The recent implementation of the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated) system in the soybean pathogen Phytophthora sojae, an experimental model for oomycetes, has opened up a powerful new research capability for the oomycete community. Here, we describe a detailed protocol for CRISPR/Cas9-mediated genome editing in P. sojae, including single guide RNA (sgRNA) design and construction, efficient gene replacement, and mutant-screening strategies. This protocol should be generally applicable for most culturable oomycetes. We also describe an optimized transformation method that is useful for other Phytophthora spp. including P. capsici and P. parasitica. © 2017 by John Wiley & Sons, Inc.
Niu, Y L; Guo, W Y; Bai, L R; Zhao, J C
Knowledge of the spatial patterns of genetic variation in wild populations has significant implications for in situ conservation and the determination of conservation order. To study the levels of genetic diversity, spatial genetic structures, and genetic distances in Glycine soja, 11 natural populations in northern China were analyzed by estimating genetic coefficients using inter-simple sequence repeat (ISSR) fingerprints via mixed sampling strategies. Sixteen ISSR primers generated 98 reproducible polymorphic amplification banding patterns of 172 scored, accounting for 56.98% of the polymorphisms among the populations. The dendrogram based on Nei's genetic distance showed that distinct genetic differentiation occurred in G. soja. The Unweighted Pair-Group Method with Arithmetic Mean cluster analysis indicated two broad groups, and one contained all of the populations except three from Chengde, which formed the smaller second group. The spatial genetic structure evident in the wild soybean populations may be attributed to restricted seed dispersal and the dominant breeding system of this species. The detection of genetic structures in wild soybean populations could be a significant index for the effective conservation of many wild populations, and it could be exploited by soybean breeding programs to increase production.
Yu, Bingjun; Ji, Xiaojia; Liu, Jun; Liu, Youliang
With internationally common-used Glycine max (the salt-tolerant Lee68) and Glycine soja (the salt-sensitive N23232) as reference, this paper studied the polyamines (PAs) contents and polyamine oxidase (PAO) activities in the highly salt-tolerant BB52 (Glycine soja) seedlings, which showed that under 150mmol x L(-1) NaCl stress for 2d, the decrease of Put and Spd contents was more significant, but that of Spd content was less significant in roots of BB52 than in those of Lee68 and N23232. For leaves, the decrease of Put and increase of Spd contents were markedly observed in BB52. The ascent of (Spm + Spd)/Put ratios and descent of Put/PAs ratios showed a positive relation to their salt tolerance. The PAO activity in roots and leaves was all increased, and most obvious in N23232. The relationship between PAs levels in BB52 and its salt tolerance was also discussed.
Lee, Jeong-Dong; Shannon, J Grover; Vuong, Tri D; Nguyen, Henry T
Tolerant soybean (Glycine max [L.] Merr.) cultivars aid in reducing salt damage in problem fields. New genes are important to reduce losses from salt injury. Objectives of this study were to determine inheritance of salt tolerance in wild soybean (Glycine soja Sieb. and Zucc.) PI483463 and to test allelism of tolerance genes from genotypes PI483463 and S-100, a common ancestor of southern in US cultivars. Tolerant (T) PI483463 was crossed to sensitive (S) cultivar Hutcheson to study inheritance. PI483463 (T) was crossed with S-100 (T) to test for allelism. Parents, F(1) plants, F(2) populations, and F(2:3) lines were assayed in a 100 mM salt solution to determine tolerance. F(2) from T x S cross segregated 3(T):1 (S) and the F(2:3) lines responded 1 (T): 2 (segregating):1 (S). F(2) plants from PI483463 (T) x S-100 (T) segregated 15 (T):1 (S) indicating different genes from the 2 sources. Results showed that G. soja line PI483463 had a single dominant gene for salt tolerance, which was different than the gene in G. max line S-100. The symbol, Ncl2, was designated for this new salt tolerance allele.
Jiang, Liangyu; Wu, Junjiang; Fan, Sujie; Li, Wenbin; Dong, Lidong; Cheng, Qun; Xu, Pengfei; Zhang, Shuzhen
Pathogenesis-related proteins (PR proteins) play crucial roles in the plant defense system. A novel PRP gene was isolated from highly resistant soybean infected with Phytophthora sojae (P. sojae) and was named GmPRP (GenBank accession number: KM506762). The amino acid sequences of GmPRP showed identities of 74%, 73%, 72% and 69% with PRP proteins from Vitis vinifera, Populus trichocarpa, Citrus sinensis and Theobroma cacao, respectively. Quantitative real-time reverse transcription PCR (qRT-PCR) data showed that the expression of GmPRP was highest in roots, followed by the stems and leaves. GmPRP expression was upregulated in soybean leaves infected with P. sojae. Similarly, GmPRP expression also responded to defense/stress signaling molecules, including salicylic acid (SA), ethylene (ET), abscisic acid (ABA) and jasmonic acid (JA). GmPRP was localized in the cell plasma membrane and cytoplasm. Recombinant GmPRP protein exhibited ribonuclease activity and significant inhibition of hyphal growth of P. sojae 1 in vitro. Overexpression of the GmPRP gene in T2 transgenic tobacco and T2 soybean plants resulted in enhanced resistance to Phytophthora nicotianae (P. nicotianae) and P. sojae race 1, respectively. These results indicated that the GmPRP protein played an important role in the defense of soybean against P. sojae infection. PMID:26114301
Yin, Weixiao; Dong, Suomeng; Zhai, Luchong; Lin, Yachun; Zheng, Xiaobo; Wang, Yuanchao
Soybean root and stem rot is caused by the oomycete pathogen Phytophthora sojae. The interaction between P. sojae and soybean fits the "gene-for-gene" hypothesis. Although more than 10 P. sojae avirulence (Avr) effectors have been genetically identified, nearly half of genetically defined avr genes have been cloned. In a previous bioinformatic and global transcriptional analysis, we identified a P. sojae RxLR effector, Avr1d, which was 125 amino acids in length. Mapping data demonstrated that Avr1d presence or absence in the genome was co-segregated with the Avr1d avirulence phenotype in F2 populations. Transient expression of the Avr1d gene using co-bombardment in soybean isogenic lines revealed that this gene triggered a hypersensitive response (HR) in the presence of Rps1d. Sequencing of Avr1d genes in different P. sojae strains revealed two Avr1d alleles. Although polymorphic, the two Avr1d alleles could trigger Rps1d-mediated HR. P. sojae strains carrying either of the alleles were avirulent on Rps1d soybean lines. Avr1d was upregulated during the germinating cyst and early infection stages. Furthermore, transient expression of Avr1d in Nicotiana benthamiana suppressed BAX-induced cell death and enhanced P. capsici infection. Avr1d also suppressed effector-triggered immunity induction by associating with Avr1b and Rps1b, suggestive of a role in suppressing plant immunity.
Ma, Zhenchuan; Song, Tianqiao; Zhu, Lin; Ye, Wenwu; Wang, Yang; Shao, Yuanyuan; Dong, Suomeng; Zhang, Zhengguang; Dou, Daolong; Zheng, Xiaobo; Tyler, Brett M.; Wang, Yuanchao
We identified a glycoside hydrolase family 12 (GH12) protein, XEG1, produced by the soybean pathogen Phytophthora sojae that exhibits xyloglucanase and β-glucanase activity. It acts as an important virulence factor during P. sojae infection but also acts as a pathogen-associated molecular pattern (PAMP) in soybean (Glycine max) and solanaceous species, where it can trigger defense responses including cell death. GH12 proteins occur widely across microbial taxa, and many of these GH12 proteins induce cell death in Nicotiana benthamiana. The PAMP activity of XEG1 is independent of its xyloglucanase activity. XEG1 can induce plant defense responses in a BAK1-dependent manner. The perception of XEG1 occurs independently of the perception of ethylene-inducing xylanase. XEG1 is strongly induced in P. sojae within 30 min of infection of soybean and then slowly declines. Both silencing and overexpression of XEG1 in P. sojae severely reduced virulence. Many P. sojae RXLR effectors could suppress defense responses induced by XEG1, including several that are expressed within 30 min of infection. Therefore, our data suggest that PsXEG1 contributes to P. sojae virulence, but soybean recognizes PsXEG1 to induce immune responses, which in turn can be suppressed by RXLR effectors. XEG1 thus represents an apoplastic effector that is recognized via the plant’s PAMP recognition machinery. PMID:26163574
Arnemann, Jonas André; Walsh, Tom K; Gordon, Karl H J; Brier, Hugh; Guedes, Jerson Vanderlei Carús; Tay, Wee Tek
We report the complete mitochondrial DNA genome of the soybean stem fly (SSF) Melanagromyza sojae from Brazil Santa Catarina state based on Illumina MiSeq sequence data. The estimated mitogenome is 15 475 base pairs (bp) (KT597923), with 13 protein-coding genes (PCGs), two ribosomal RNA (rRNA) genes, and 22 tRNAs, and an estimated 579 bp AT-rich control region. Similar to other insects, the SSF mitogenome is A-T bias with 40.9% A, 36.7% T, 13.6% C, and 8.8% G. Molecular characterization of SSF mitogenome will facilitate the development of effective molecular markers, and robust and rapid identification of suspected biosecurity incursions and field infestations of this insect pest.
Kim, Hyo Jung; Suh, Hwa-Jin; Kim, Jeong Hwan; Kang, Sun Chul; Park, Sunmin; Lee, Choong Hwan; Kim, Jong-Sang
Glyceollins, which are synthesized from daidzein in soybeans cultured with fungi, have been shown to have antifungal effects and cancer preventive properties. Several studies have proposed that isoflavones and their metabolites act as a mixed agonist/antagonist for estrogen. Although glyceollins were reported to suppress some cancer cells via anti-estrogenic activity, it is not clear whether the compounds possess estrogenic potential. In contrast to the anti-estrogenic action reported thus far, we observed estrogenic effects of glyceollins using E-screen assay and pS2 expression, whereas glyceollins showed higher affinity for estrogen receptor (ER) beta than ERalpha. We also found that glyceollins were more efficiently produced de novo in minced than in half-sliced soybean, following infection with Aspergillus sojae. In conclusion, glyceollins may be useful in the prevention or amelioration of postmenopausal complications because they had strong estrogenic activity, and their production could be variable depending upon processing prior to fungal inoculation.
Zhu, Dan; Bai, Xi; Luo, Xiao; Chen, Qin; Cai, Hua; Ji, Wei; Zhu, Yanming
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.
Fan, Sujie; Jiang, Liangyu; Wu, Junjiang; Dong, Lidong; Cheng, Qun; Xu, Pengfei; Zhang, Shuzhen
Phytophthora root and stem rot of soybean, caused by Phytophthora sojae (P. sojae), is a destructive disease in many soybean planting regions worldwide. In a previous study, an expressed sequence tag (EST) homolog of the major allergen Pru ar 1 in apricot (Prunus armeniaca) was identified up-regulated in the highly resistant soybean ‘Suinong 10’ infected with P. sojae. Here, the full length of the EST was isolated using rapid amplification of cDNA ends (RACE). It showed the highest homolgy of 53.46% with Gly m 4 after comparison with the eight soybean allergen families reported and was named Gly m 4-like (Gly m 4l, GenBank accession no. HQ913577.1). The cDNA full length of Gly m 4l was 707 bp containing a 474 bp open reading frame encoding a polypeptide of 157 amino acids. Sequence analysis suggests that Gly m 4l contains a conserved ‘P-loop’ (phosphate-binding loop) motif at residues 47–55 aa and a Bet v 1 domain at residues 87–120 aa. The transcript abundance of Gly m 4l was significantly induced by P. sojae, salicylic acid (SA), NaCl, and also responded to methyl jasmonic acid (MeJA) and ethylene (ET). The recombinant Gly m 4l protein showed RNase activity and displayed directly antimicrobial activity that inhibited hyphal growth and reduced zoospore release in P. sojae. Further analyses showed that the RNase activity of the recombinant protein to degrading tRNA was significantly affected in the presence of zeatin. Over-expression of Gly m 4l in susceptible ‘Dongnong 50’ soybean showed enhanced resistance to P. sojae. These results indicated that Gly m 4l protein played an important role in the defense of soybean against P. sojae infection. PMID:26474489
Luo, Qingyun; Yu, Bingjun; Liu, Youliang
High Na+ and Cl- concentrations in soil cause hyperionic and hyperosmotic stress effects, the consequence of which can be plant demise. Ion-specific stress effects of Na+ and Cl- on seedlings of cultivated (Glycine max (L.) Merr) and wild soybean (Glycine soja Sieb. Et Zucc.) were evaluated and compared in isoosmotic solutions of Cl-, Na+ and NaCl. Results showed that under NaCl stress, Cl- was more toxic than Na+ to seedlings of G. max. Injury of six G. max cultivars, including 'Jackson' (salt sensitive) and 'Lee 68' (salt tolerant), was positively correlated with the content of Cl- in the leaves, and negatively with that in the roots. In subsequent research, seedlings of two G. max cultivars (salt-tolerant Nannong 1138-2, and salt-sensitive Zhongzihuangdou-yi) and two G. soja populations (BB52 and N23232) were subjected to isoosmotic solutions of 150mM Na+, Cl- and NaCl, respectively. G. max cv. Nannong 1138-2 and Zhongzihuangdou-yi were damaged much more heavily in the solution of Cl- than in that of Na+. Their Leaves were found to be more sensitive to Cl- than to Na+, and salt tolerance of these two G. max cultivars was mainly due to successful withholding of Cl- in the roots and stems to decrease its content in the leaves. The reverse response to isoosmotic stress of 150 mM Na+ and Cl- was shown in G. soja populations of BB52 and N23232; their leaves were not as susceptible to toxicity of Cl- as that of Na+. Salt tolerance of BB52 and N23232 was mainly due to successful withholding of Na+ in the roots and stems to decrease its content in the leaves. These results indicate that G. soja have advantages over G. max in those traits associated with the mechanism of Cl-tolerance, such as its withholding in roots and vacuoles of leaves. It is possible to use G. soja to improve the salt tolerance of G. max.
Li, Ying-Hui; Li, Wei; Zhang, Chen; Yang, Liang; Chang, Ru-Zhen; Gaut, Brandon S; Qiu, Li-Juan
• The study of genetic diversity between a crop and its wild relatives may yield fundamental insights into evolutionary history and the process of domestication. • In this study, we genotyped a sample of 303 accessions of domesticated soybean (Glycine max) and its wild progenitor Glycine soja with 99 microsatellite markers and 554 single-nucleotide polymorphism (SNP) markers. • The simple sequence repeat (SSR) loci averaged 21.5 alleles per locus and overall Nei's gene diversity of 0.77. The SNPs had substantially lower genetic diversity (0.35) than SSRs. A SSR analyses indicated that G. soja exhibited higher diversity than G. max, but SNPs provided a slightly different snapshot of diversity between the two taxa. For both marker types, the primary division of genetic diversity was between the wild and domesticated accessions. Within taxa, G. max consisted of four geographic regions in China. G. soja formed six subgroups. Genealogical analyses indicated that cultivated soybean tended to form a monophyletic clade with respect to G. soja. • G. soja and G. max represent distinct germplasm pools. Limited evidence of admixture was discovered between these two species. Overall, our analyses are consistent with the origin of G. max from regions along the Yellow River of China.
Heerd, Doreen; Diercks-Horn, Sonja; Fernández-Lahore, Marcelo
Previously identified fungal pectinase producers of the species Aspergillus sojae were used for optimization of polygalacturonase production in solid-state fermentation applying Design of Experiment. The effects of media composition and several process parameters, like inoculum size, moisture level, incubation time and temperature on polygalacturonase activity were studied in screening and optimization investigations. Utilization of agricultural and agro-industrial by-products provided the establishment of a cost-efficient and sustainable process for enzyme production. Comparison of pectinase production by A. sojae ATCC 20235 and A. sojae CBS 100928 under optimized conditions yielded 6.9 times higher polygalacturonase activity by A. sojae ATCC 20235. Highest enzyme yield (909.5 ± 2.7 U/g) was obtained by A. sojae ATCC 20235 after 8 days at 30°C applying 30% sugar beet pulp as inducer substrate in combination with wheat bran as medium wetted at 160% with 0.2 M HCl. Furthermore, an overview of pectinolytic enzyme activities present in the extracts of both strains is provided. Protein profiles of both strains are given by SDS-PAGE electrophoresis, as well as zymograms for pectinolytic enzymes in comparison to commercial pectinase preparations.
Wu, Li Juan; Wang, Hai Qing; Wang, En Tao; Chen, Wen Xin; Tian, Chang Fu
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.
Nedukha, O M; Leach, J E; Ryba-White, M; Hilaire, E; Guikema, J; Kordyum, E L
The study of pathogenicity of higher plants under conditions of microgravity is of great importance for the future production of food in space. Previous work suggests that microgravity affects both microbes and plants. Bacterial numbers increased after 17 days in an algae-bacterium association on the biosatellite "Kosmos-1887". This was speculated to result from an increase in the multiplication rate of the bacteria. Sporangia of both Actinomices brevis, in the shuttles "Soyuz-19" and "Appolon", and Phycomyces blakes, in biosatellite "Kosmos-936", formed after 10 days in microgravity. Sporangia did not form in the ground controls in the same time suggesting that the rate of fungal development is enhanced in microgravity. Plant responses to pathogens in microgravity have not been studied, however, microgravity profoundly impacts plant cell development, cytology, and physiology. In microgravity, developing cell walls are thinner and contain less lignin than ground-grown plants. The demonstrated effects of microgravity on both plants and microbes lead us to hypothesize that plants may be more susceptible to pathogens under conditions of microgravity. The aim of this study was to determine the influence of microgravity on the susceptibility of soybean to the fungal root rot pathogen, Phytophthora sojae.
Zhang, Meixiang; Ahmed Rajput, Nasir; Shen, Danyu; Sun, Peng; Zeng, Wentao; Liu, Tingli; Juma Mafurah, Joseph; Dou, Daolong
Each oomycete pathogen encodes a large number of effectors. Some effectors can be used in crop disease resistance breeding, such as to accelerate R gene cloning and utilisation. Since cytoplasmic effectors may cause acute physiological changes in host cells at very low concentrations, we assume that some of these effectors can serve as functional genes for transgenic plants. Here, we generated transgenic Nicotiana benthamiana plants that express a Phytophthora sojae CRN (crinkling and necrosis) effector, PsCRN115. We showed that its expression did not significantly affect the growth and development of N. benthamiana, but significantly improved disease resistance and tolerance to salt and drought stresses. Furthermore, we found that expression of heat-shock-protein and cytochrome-P450 encoding genes were unregulated in PsCRN115-transgenic N. benthamiana based on digital gene expression profiling analyses, suggesting the increased plant defence may be achieved by upregulation of these stress-related genes in transgenic plants. Thus, PsCRN115 may be used to improve plant tolerance to biotic and abiotic stresses.
Chen, Xuan-Qin; Yu, Bing-Jun; Liu, You-Liang
The seedlings of the F4 hybrid strain 'JB185' selected for salt tolerance generation by generation, their parents Glycine max cv. Jackson and Glycine soja population 'BB52' were treated with different NaCl concentrations and iso-osmotic (-0.53 MPa) PEG-6000, NaCl, Na+ (without Cl-) and Cl- (without Na+) solutions for 6 d. The results showed that: (1) The relative electrolyte leakage and malondialdehyde (MDA) content in leaves of the above three soybean seedlings showed an increase trend when the NaCl concentration was elevated, but chlorophyll contents decreased except the significant increase in 'BB52' and 'JB185' under NaCl 50 mmol/L stress. The change in 'JB185' was between its parents. (2) Under different iso-osmotic stresses, the relative electrolyte leakage and MDA contents in leaves of three soybean seedlings also increased mostly, the changes in 'BB52' and 'JB185' under Na+ (without Cl-) stress were more than those under Cl- (without Na+) stress. The free and bound Put, Spd and Spm contents in leaves all increased when compared with the control, the ratios of free (Spd+Spm)/Put and total bound polyamines in 'BB52' and 'JB185' seedlings under Na+ (without Cl-) treatment were the lowest one among three iso-osmotic salt stresses. The results indicate that the F4 hybrid strain 'JB185' is more sensitive to Na+ than Cl- as its wild parent 'BB52' population.
Zhu, Dan; Cai, Hua; Luo, Xiao; Bai, Xi; Deyholos, Michael K.; Chen, Qin; Chen, Chao; Ji, Wei; Zhu, Yanming
Highlights: Black-Right-Pointing-Pointer We isolated and characterized a novel JAZ family gene, GsJAZ2, from Glycine soja. Black-Right-Pointing-Pointer Overexpression of GsJAZ2 enhanced plant tolerance to salt and alkali stress. Black-Right-Pointing-Pointer The transcriptions of stress marker genes were higher in GsJAZ2 overexpression lines. Black-Right-Pointing-Pointer GsJAZ2 was localized to nucleus. -- Abstract: Salt and alkali stress are two of the main environmental factors limiting crop production. Recent discoveries show that the JAZ family encodes plant-specific genes involved in jasmonate signaling. However, there is only limited information about this gene family in abiotic stress response, and in wild soybean (Glycine soja), which is a species noted for its tolerance to alkali and salinity. Here, we isolated and characterized a novel JAZ family gene, GsJAZ2, from G. soja. Transcript abundance of GsJAZ2 increased following exposure to salt, alkali, cold and drought. Over-expression of GsJAZ2 in Arabidopsis resulted in enhanced plant tolerance to salt and alkali stress. The expression levels of some alkali stress response and stress-inducible marker genes were significantly higher in the GsJAZ2 overexpression lines as compared to wild-type plants. Subcellular localization studies using a GFP fusion protein showed that GsJAZ2 was localized to the nucleus. These results suggest that the newly isolated wild soybean GsJAZ2 is a positive regulator of plant salt and alkali stress tolerance.
Kyrychenko, A M
This paper presents data on BYMV effects on some physiological processes of Glycine soja L. cultivated in the right-bank forest-steppe regions. Pigment content (chlorophyll a, b and carotenoids), soluble proteins and water soluble carbohydrates were estimated and, as has been shown, are subjected to significant changes as compared with control plants, namely: a decrease in the content of chlorophyll a, b and carotenoids was 64%, 53% and 36% compared with the control plants. The significant increase in carbohydrates (56% compared to the control) was observed at the end of the test period.
Sun, Yuzhe; Mui, Zeta; Liu, Xuan; Yim, Aldrin Kay-Yuen; Qin, Hao; Wong, Fuk-Ling; Chan, Ting-Fung; Yiu, Siu-Ming; Lam, Hon-Ming; Lim, Boon Leong
Small RNAs, including microRNAs (miRNAs) and phased small interfering RNAs (phasiRNAs; from PHAS loci), play key roles in plant development. Cultivated soybean, Glycine max, contributes a great deal to food production, but, compared to its wild kin, Glycine soja, it may lose some genetic information during domestication. In this work, we analyzed the sRNA profiles of different tissues in both cultivated (C08) and wild soybeans (W05) at three stages of development. A total of 443 known miRNAs and 15 novel miRNAs showed varying abundances between different samples, but the miRNA profiles were generally similar in both accessions. Based on a sliding window analysis workflow that we developed, 50 PHAS loci generating 55 21-nucleotide phasiRNAs were identified in C08, and 46 phasiRNAs from 41 PHAS loci were identified in W05. In germinated seedlings, phasiRNAs were more abundant in C08 than in W05. Disease resistant TIR-NB-LRR genes constitute a very large family of PHAS loci. PhasiRNAs were also generated from several loci that encode for NAC transcription factors, Dicer-like 2 (DCL2), Pentatricopeptide Repeat (PPR), and Auxin Signaling F-box 3 (AFB3) proteins. To investigate the possible involvement of miRNAs in initiating the PHAS-phasiRNA pathway, miRNA target predictions were performed and 17 C08 miRNAs and 15 W05 miRNAs were predicted to trigger phasiRNAs biogenesis. In summary, we provide a comprehensive description of the sRNA profiles of wild versus cultivated soybeans, and discuss the possible roles of sRNAs during soybean germination. PMID:27929436
Kuroda, Y; Kaga, A; Tomooka, N; Vaughan, D A
The research objectives were to determine aspects of the population dynamics relevant to effective monitoring of gene flow in the soybean crop complex in Japan. Using 20 microsatellite primers, 616 individuals from 77 wild soybean (Glycine soja) populations were analysed. All samples were of small seed size (< 0.03 g), were directly collected in the field and came from all parts of Japan where wild soybeans grow, except Hokkaido. Japanese wild soybean showed significant reduction in observed heterozygosity, low outcrossing rate (mean 3.4%) and strong genetic differentiation among populations. However, the individual assignment test revealed evidence of rare long-distance seed dispersal (> 10 km) events among populations, and spatial autocorrelation analysis revealed that populations within a radius of 100 km showed a close genetic relationship to one another. When analysis of graphical ordination was applied to compare the microsatellite variation of wild soybean with that of 53 widely grown Japanese varieties of cultivated soybean (Glycine max), the primary factor of genetic differentiation was based on differences between wild and cultivated soybeans and the secondary factor was geographical differentiation of wild soybean populations. Admixture analysis revealed that 6.8% of individuals appear to show introgression from cultivated soybeans. These results indicated that population genetic structure of Japanese wild soybean is (i) strongly affected by the founder effect due to seed dispersal and inbreeding strategy, (ii) generally well differentiated from cultivated soybean, but (iii) introgression from cultivated soybean occurs. The implications of the results for the release of transgenic soybeans where wild soybeans grow are discussed.
Fukuda, Takako; Maruyama, Nobuyuki; Kanazawa, Akira; Abe, Jun; Shimamoto, Yoshiya; Hiemori, Miki; Tsuji, Hideaki; Tanisaka, Takatoshi; Utsumi, Shigeru
Cultivated soybeans (Glycine max) are derived from wild soybeans (Glycine soja) and can be crossed with them to produce fertile offspring. The latter exhibit greater genetic variation than the former, suggesting a possibility that wild soybeans contain storage proteins with properties different from and better than those of cultivated soybeans. To identify a wild soybean suitable for breeding a new soybean cultivar, we analyzed seed proteins from 390 lines of wild soybeans by electrophoresis. We found some lines containing electrophoretic variants of glycinin and beta-conglycinin subunits: one line containing a small alpha' subunit of beta-conglycinin and two and five lines containing small A3 and large A4 polypeptides of glycinin, respectively. Beta-Conglycinin and glycinin containing such variant subunits exhibited solubility and emulsifying ability similar to those of the predominant types of wild and cultivated soybeans. Glycinins containing small A3 and large A4 gave a shoulder derived from the start of denaturation at a temperature 4 degrees C lower than that of glycinin from the predominant types of wild and cultivated soybeans, although their thermal denaturation midpoint temperatures were very similar to each other. Cloning and sequencing of the predominant and variant subunit cDNAs revealed that the small alpha' and the small A3 lacked 24 amino acid residues in the extension region and four amino acid residues in the hypervariable region, respectively, and that the large A4 did not have an insert corresponding to the difference in the electrophoretic mobility but Arg279 and Gln305 were replaced by glutamine and histidine, respectively, in the hypervariable region. These suggest that small differences even in the hypervariable region can affect the thermal stability, as well as the electrophoretic mobilities, of the proteins.
Sun, Yuzhe; Mui, Zeta; Liu, Xuan; Yim, Aldrin Kay-Yuen; Qin, Hao; Wong, Fuk-Ling; Chan, Ting-Fung; Yiu, Siu-Ming; Lam, Hon-Ming; Lim, Boon Leong
Small RNAs, including microRNAs (miRNAs) and phased small interfering RNAs (phasiRNAs; from PHAS loci), play key roles in plant development. Cultivated soybean, Glycine max, contributes a great deal to food production, but, compared to its wild kin, Glycine soja, it may lose some genetic information during domestication. In this work, we analyzed the sRNA profiles of different tissues in both cultivated (C08) and wild soybeans (W05) at three stages of development. A total of 443 known miRNAs and 15 novel miRNAs showed varying abundances between different samples, but the miRNA profiles were generally similar in both accessions. Based on a sliding window analysis workflow that we developed, 50 PHAS loci generating 55 21-nucleotide phasiRNAs were identified in C08, and 46 phasiRNAs from 41 PHAS loci were identified in W05. In germinated seedlings, phasiRNAs were more abundant in C08 than in W05. Disease resistant TIR-NB-LRR genes constitute a very large family of PHAS loci. PhasiRNAs were also generated from several loci that encode for NAC transcription factors, Dicer-like 2 (DCL2), Pentatricopeptide Repeat (PPR), and Auxin Signaling F-box 3 (AFB3) proteins. To investigate the possible involvement of miRNAs in initiating the PHAS-phasiRNA pathway, miRNA target predictions were performed and 17 C08 miRNAs and 15 W05 miRNAs were predicted to trigger phasiRNAs biogenesis. In summary, we provide a comprehensive description of the sRNA profiles of wild versus cultivated soybeans, and discuss the possible roles of sRNAs during soybean germination.
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...
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-...
Fang, Yufeng; Tyler, Brett M
Oomycetes are fungal-like eukaryotic microbes in the kingdom Stramenopila. We recently found that the oomycete plant pathogen Phytophthora sojae uses nuclear localization signals (NLSs) for translocation of proteins into the nucleus that differ from conventional well-characterized NLSs from mammals and yeast. Here we have characterized in depth the nuclear localization signals of a P. sojae basic leucine zipper transcription factor, PsbZIP1. Nuclear localization of PsbZIP1 was determined by a central conserved region overlapping the DNA binding domain. Mutational analysis of this region identified four distinct elements that contributed multiplicatively to nuclear localization, but the conserved DNA binding residues were not required. Three of the elements showed autonomous NLS activity and the fourth served as a nuclear localization enhancer. Sequences within two of the nuclear localization elements defined a new form of bipartite NLS consisting of a triplet of basic residues followed by a tail of scattered basic amino acids. This article is protected by copyright. All rights reserved.
Leamy, Larry J; Lee, Cheng-Ruei; Song, Qijian; Mujacic, Ibro; Luo, Yan; Chen, Charles Y; Li, Changbao; Kjemtrup, Susanne; Song, Bao-Hua
A fundamental goal in evolutionary biology is to understand how various evolutionary factors interact to affect the population structure of diverse species, especially those of ecological and/or agricultural importance such as wild soybean (Glycine soja). G. soja, from which domesticated soybeans (Glycine max) were derived, is widely distributed throughout diverse habitats in East Asia (Russia, Japan, Korea, and China). Here, we utilize over 39,000 single nucleotide polymorphisms genotyped in 99 ecotypes of wild soybean sampled across their native geographic range in northeast Asia, to understand population structure and the relative contribution of environment versus geography to population differentiation in this species. A STRUCTURE analysis identified four genetic groups that largely corresponded to the geographic regions of central China, northern China, Korea, and Japan, with high levels of admixture between genetic groups. A canonical correlation and redundancy analysis showed that environmental factors contributed 23.6% to population differentiation, much more than that for geographic factors (6.6%). Precipitation variables largely explained divergence of the groups along longitudinal axes, whereas temperature variables contributed more to latitudinal divergence. This study provides a foundation for further understanding of the genetic basis of climatic adaptation in this ecologically and agriculturally important species.
Zhu, Dan; Cai, Hua; Luo, Xiao; Bai, Xi; Deyholos, Michael K; Chen, Qin; Chen, Chao; Ji, Wei; Zhu, Yanming
Salt and alkali stress are two of the main environmental factors limiting crop production. Recent discoveries show that the JAZ family encodes plant-specific genes involved in jasmonate signaling. However, there is only limited information about this gene family in abiotic stress response, and in wild soybean (Glycine soja), which is a species noted for its tolerance to alkali and salinity. Here, we isolated and characterized a novel JAZ family gene, GsJAZ2, from G. soja. Transcript abundance of GsJAZ2 increased following exposure to salt, alkali, cold and drought. Over-expression of GsJAZ2 in Arabidopsis resulted in enhanced plant tolerance to salt and alkali stress. The expression levels of some alkali stress response and stress-inducible marker genes were significantly higher in the GsJAZ2 overexpression lines as compared to wild-type plants. Subcellular localization studies using a GFP fusion protein showed that GsJAZ2 was localized to the nucleus. These results suggest that the newly isolated wild soybean GsJAZ2 is a positive regulator of plant salt and alkali stress tolerance.
Li, Delong; Zhao, Zhijian; Huang, Yidan; Lu, Zhaojun; Yao, Meng; Hao, Yujuan; Zhai, Chunhua; Wang, Yuanchao
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.
Anderson, Justin E.; Kono, Thomas J. Y.; Stupar, Robert M.; Kantar, Michael B.; Morrell, Peter L.
Natural populations across a species range demonstrate population structure owing to neutral processes such as localized origins of mutations and migration limitations. Selection also acts on a subset of loci, contributing to local adaptation. An understanding of the genetic basis of adaptation to local environmental conditions is a fundamental goal in basic biological research. When applied to crop wild relatives, this same research provides the opportunity to identify adaptive genetic variation that may be used to breed for crops better adapted to novel or changing environments. The present study explores an ex situ conservation collection, the USDA germplasm collection, genotyped at 32,416 SNPs to identify population structure and test for associations with bioclimatic and biophysical variables in Glycine soja, the wild progenitor of Glycine max (soybean). Candidate loci were detected that putatively contribute to adaptation to abiotic stresses. The identification of potentially adaptive variants in this ex situ collection may permit a more targeted use of germplasm collections. PMID:26818076
Chai, Chuan; Cui, Xiaobing; Shan, Chenxiao; Yu, Sheng; Wen, Hongmei
UFLC-QTRAP MS was used to develop a sensitive and rapid method of evaluating content variation during Semen sojae praeparatum (SSP) fermentation. It did this through the simultaneous quantification of 22 free amino acids (FAAs) and 16 nucleosides and nucleobases (Ns) in the raw materials and processed products of SSP. The method was shown to be reproducible and accurate. The limits of detection (LOD) and quantity (LOQ) values were between 0.09-168.75 and 0.31-562.50 ng/mL for the 38 analytes, respectively. The data was examined through Principal Component Analysis (PCA) to compare the content variations. The quantitative results showed that the ingredients were properly determined in most of the samples and were converted regularly throughout the SSP fermentation process. These results correspond to the morphologic changes and PCA results.
Anderson, Justin E; Kono, Thomas J Y; Stupar, Robert M; Kantar, Michael B; Morrell, Peter L
Natural populations across a species range demonstrate population structure owing to neutral processes such as localized origins of mutations and migration limitations. Selection also acts on a subset of loci, contributing to local adaptation. An understanding of the genetic basis of adaptation to local environmental conditions is a fundamental goal in basic biological research. When applied to crop wild relatives, this same research provides the opportunity to identify adaptive genetic variation that may be used to breed for crops better adapted to novel or changing environments. The present study explores an ex situ conservation collection, the USDA germplasm collection, genotyped at 32,416 SNPs to identify population structure and test for associations with bioclimatic and biophysical variables in Glycine soja, the wild progenitor of Glycine max (soybean). Candidate loci were detected that putatively contribute to adaptation to abiotic stresses. The identification of potentially adaptive variants in this ex situ collection may permit a more targeted use of germplasm collections.
Jing, Maofeng; Guo, Baodian; Li, Haiyang; Yang, Bo; Wang, Haonan; Kong, Guanghui; Zhao, Yao; Xu, Huawei; Wang, Yan; Ye, Wenwu; Dong, Suomeng; Qiao, Yongli; Tyler, Brett M.; Ma, Wenbo; Wang, Yuanchao
Phytophthora pathogens secrete an array of specific effector proteins to manipulate host innate immunity to promote pathogen colonization. However, little is known about the host targets of effectors and the specific mechanisms by which effectors increase susceptibility. Here we report that the soybean pathogen Phytophthora sojae uses an essential effector PsAvh262 to stabilize endoplasmic reticulum (ER)-luminal binding immunoglobulin proteins (BiPs), which act as negative regulators of plant resistance to Phytophthora. By stabilizing BiPs, PsAvh262 suppresses ER stress-triggered cell death and facilitates Phytophthora infection. The direct targeting of ER stress regulators may represent a common mechanism of host manipulation by microbes. PMID:27256489
Dong, Suomeng; Yin, Weixiao; Kong, Guanghui; Yang, Xinyu; Qutob, Dinah; Chen, Qinghe; Kale, Shiv D.; Sui, Yangyang; Zhang, Zhengguang; Dou, Daolong; Zheng, Xiaobo; Gijzen, Mark; M. Tyler, Brett; Wang, Yuanchao
Plants have evolved pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI) to protect themselves from infection by diverse pathogens. Avirulence (Avr) effectors that trigger plant ETI as a result of recognition by plant resistance (R) gene products have been identified in many plant pathogenic oomycetes and fungi. However, the virulence functions of oomycete and fungal Avr effectors remain largely unknown. Here, we combined bioinformatics and genetics to identify Avr3b, a new Avr gene from Phytophthora sojae, an oomycete pathogen that causes soybean root rot. Avr3b encodes a secreted protein with the RXLR host-targeting motif and C-terminal W and Nudix hydrolase motifs. Some isolates of P. sojae evade perception by the soybean R gene Rps3b through sequence mutation in Avr3b and lowered transcript accumulation. Transient expression of Avr3b in Nicotiana benthamiana increased susceptibility to P. capsici and P. parasitica, with significantly reduced accumulation of reactive oxygen species (ROS) around invasion sites. Biochemical assays confirmed that Avr3b is an ADP-ribose/NADH pyrophosphorylase, as predicted from the Nudix motif. Deletion of the Nudix motif of Avr3b abolished enzyme activity. Mutation of key residues in Nudix motif significantly impaired Avr3b virulence function but not the avirulence activity. Some Nudix hydrolases act as negative regulators of plant immunity, and thus Avr3b might be delivered into host cells as a Nudix hydrolase to impair host immunity. Avr3b homologues are present in several sequenced Phytophthora genomes, suggesting that Phytophthora pathogens might share similar strategies to suppress plant immunity. PMID:22102810
Sun, Xiaoli; Sun, Mingzhe; Jia, Bowei; Chen, Chao; Qin, Zhiwei; Yang, Kejun; Shen, Yang; Meiping, Zhang; Mingyang, Cong; Zhu, Yanming
It is widely accepted that the 14-3-3 family proteins are key regulators of multiple stress signal transduction cascades. By conducting genome-wide analysis, researchers have identified the soybean 14-3-3 family proteins; however, until now, there is still no direct genetic evidence showing the involvement of soybean 14-3-3s in ABA responses. Hence, in this study, based on the latest Glycine max genome on Phytozome v10.3, we initially analyzed the evolutionary relationship, genome organization, gene structure and duplication, and three-dimensional structure of soybean 14-3-3 family proteins systematically. Our results suggested that soybean 14-3-3 family was highly evolutionary conserved and possessed segmental duplication in evolution. Then, based on our previous functional characterization of a Glycine soja 14-3-3 protein GsGF14o in drought stress responses, we further investigated the expression characteristics of GsGF14o in detail, and demonstrated its positive roles in ABA sensitivity. Quantitative real-time PCR analyses in Glycine soja seedlings and GUS activity assays in PGsGF14O:GUS transgenic Arabidopsis showed that GsGF14o expression was moderately and rapidly induced by ABA treatment. As expected, GsGF14o overexpression in Arabidopsis augmented the ABA inhibition of seed germination and seedling growth, promoted the ABA induced stomata closure, and up-regulated the expression levels of ABA induced genes. Moreover, through yeast two hybrid analyses, we further demonstrated that GsGF14o physically interacted with the AREB/ABF transcription factors in yeast cells. Taken together, results presented in this study strongly suggested that GsGF14o played an important role in regulation of ABA sensitivity in Arabidopsis.
Liu, Haoran; Song, Jinhui; Dong, Lijun; Wang, Di; Zhang, Shuling; Liu, Jianfeng
Understanding the mechanism for salt tolerance in wild soybean (Glycine soja) can help researchers improve that trait in cultivated soybean lines. We analyzed the effects of excess NaCl on the growth, physiology, and ion distribution in three soybean species: wild G. soja (W8), semi-wild G. gracilis (SW18), and the cultivated salt-sensitive G. max (cv. Melrose). These comparisons revealed that, under salt stress, shoot and root lengths and biomass (either shoot or root dry weights) were significantly higher for the W8 genotype than for the other two. Most of the morphological parameters for roots from the W8 plants were also increased, including total length, specific root length, and surface area. However, the average root diameter for W8 was significantly lower than that of either SW18 or 'Melrose' soybeans. In response to salinity, photosynthesis was suppressed to a greater extent in 'Melrose' than in W8. The relatively higher tolerance shown by W8 plants was also associated with lower levels of malondialdehyde, hydrogen peroxide, and relative electrical conductivity, but higher activities by superoxide dismutase and peroxidase, as well as more free proline and glycine betaine. In addition, the W8 plants contained less Na(+) and Cl(-), but more K(+), and they had a higher K(+)/Na(+) ratio in their leaves and roots when compared with either SW18 or 'Melrose' plants. Therefore, the W8 genotype performs better in terms of seedling growth, photosynthetic characteristics, and physiological indexes. These findings provide guidance for developing new soybean cultivars with improved tolerance to salt stress. Our data also contribute to the knowledge base for plant salt tolerance as a tool for increasing the yields of other crops in high-salinity soils.
Ge, Ying; Li, Yong; Lv, De-Kang; Bai, Xi; Ji, Wei; Cai, Hua; Wang, Ao-Xue; Zhu, Yan-Ming
Transcriptome of Glycine soja leaf tissue during a detailed time course formed a foundation for examining transcriptional processes during NaHCO(3) stress treatment. Of a total of 2,310 detected differentially expressed genes, 1,664 genes were upregulated and 1,704 genes were downregulated at various time points. The number of stress-regulated genes increased dramatically after a 6-h stress treatment. GO category gene enrichment analysis revealed that most of the differentially expressed genes were involved in cell structure, protein synthesis, energy, and secondary metabolism. Another enrichment test revealed that the response of G. soja to NaHCO(3) highlights specific transcription factors, such as the C2C2-CO-like, MYB-related, WRKY, GARP-G2-like, and ZIM families. Co-expressed genes were clustered into ten classes (P < 0.001). Intriguingly, one cluster of 188 genes displayed a unique expression pattern that increases at an early stage (0.5 and 3 h), followed by a decrease from 6 to 12 h. This group was enriched in regulation of transcription components, including AP2-EREBP, bHLH, MYB/MYB-related, C2C2-CO-like, C2C2-DOF, C2C2, C3H, and GARP-G2-like transcription factors. Analysis of the 1-kb upstream regions of transcripts displaying similar changes in abundance identified 19 conserved motifs, potential binding sites for transcription factors. The appearance of ABA-responsive elements in the upstream of co-expression genes reveals that ABA-mediated signaling participates in the signal transduction in alkaline response.
Wen, Zixiang; Ding, Yanlai; Zhao, Tuanjie; Gai, Junyi
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.
Tikhonov, A V; Martynov, V V; Dorokhov, D B
A comparative study of the genetic structure of natural and anthropogenic populations of G. soja gives significant information about formation of different populations, and allows developing measures for preservation of unique natural gene bank of wild soybean, the species closely related to cultivated soybean. In this study, ISSR markers were used to carry out a comparative analysis of genetic structure of natural and anthropogenic subpopulations of G. soja for studying possible mutual influence of subpopulations of anthropogenic and natural phytocenosis on the formation of their genetic diversity and to study genetic structure of natural subpopulations of wild soybean in the contact places between the two types ofcenoses. As a result, the characteristics that describe the genetic diversity of studied populations have been identified and the important role of an interaction between subpopulations of different phytocenoses on formation of the spatial genetic structure of population in the valley of Tsukanovka river has been demonstrated.
Yang, Liang; Wu, Kangcheng; Gao, Peng; Liu, Xiaojuan; Li, Guangpu; Wu, Zujian
Plant LRR-RLKs serve as protein interaction platforms, and as regulatory modules of protein activation. Here, we report the isolation of a novel plant-specific LRR-RLK from Glycine soja (termed GsLRPK) by differential screening. GsLRPK expression was cold-inducible and shows Ser/Thr protein kinase activity. Subcellular localization studies using GFP fusion protein indicated that GsLRPK is localized in the plasma membrane. Real-time PCR analysis indicated that temperature, salt, drought, and ABA treatment can alter GsLRPK gene transcription in G. soja. However, just protein induced by cold stress not by salinity and ABA treatment in tobacco was found to possess kinase activity. Furthermore, we found that overexpression of GsLRPK in yeast and Arabidopsis can enhance resistance to cold stress and increase the expression of a number of cold responsive gene markers.
Ji, Wei; Zhu, Yanming; Li, Yong; Yang, Liang; Zhao, Xiaowen; Cai, Hua; Bai, Xi
Glycine soja is a species of soybean that survives in adverse environments including high salt and drought conditions. We constructed a cDNA library from G. soja seedlings treated with NaCl and isolated a glutathione S-transferase gene (GsGST: GQ265911) from the library. The cDNA encoding GsGST contains an open reading frame of 660 bp and the predicted protein belongs to the tau class of GST family proteins. Tobacco plants over-expressing the GsGST gene showed sixfold higher GST activity than wild-type plants. Transgenic tobacco plants exhibited enhanced dehydration tolerance. T(2) transgenic tobacco plants showed higher tolerance at the seedling stage than wild-type plants to salt and mannitol as demonstrated by longer root length and less growth retardation.
Dong, Lidong; Cheng, Yingxin; Wu, Junjiang; Cheng, Qun; Li, Wenbin; Fan, Sujie; Jiang, Liangyu; Xu, Zhaolong; Kong, Fanjiang; Zhang, Dayong; Xu, Pengfei; Zhang, Shuzhen
Phytophthora root and stem rot of soybean [Glycine max (L.) Merr.], caused by Phytophthora sojae Kaufmann and Gerdemann, is a destructive disease throughout the soybean planting regions in the world. Here, we report insights into the function and underlying mechanisms of a novel ethylene response factor (ERF) in soybean, namely GmERF5, in host responses to P. sojae. GmERF5-overexpressing transgenic soybean exhibited significantly enhanced resistance to P. sojae and positively regulated the expression of the PR10, PR1-1, and PR10-1 genes. Sequence analysis suggested that GmERF5 contains an AP2/ERF domain of 58 aa and a conserved ERF-associated amphiphilic repression (EAR) motif in its C-terminal region. Following stress treatments, GmERF5 was significantly induced by P. sojae, ethylene (ET), abscisic acid (ABA), and salicylic acid (SA). The activity of the GmERF5 promoter (GmERF5P) was upregulated in tobacco leaves with ET, ABA, Phytophthora nicotianae, salt, and drought treatments, suggesting that GmERF5 could be involved not only in the induced defence response but also in the ABA-mediated pathway of salt and drought tolerance. GmERF5 could bind to the GCC-box element and act as a repressor of gene transcription. It was targeted to the nucleus when transiently expressed in Arabidopsis protoplasts. GmERF5 interacted with a basic helix-loop-helix transcription factor (GmbHLH) and eukaryotic translation initiation factor (GmEIF) both in yeast cells and in planta. To the best of our knowledge, GmERF5 is the first soybean EAR motif-containing ERF transcription factor demonstrated to be involved in the response to pathogen infection.
Zhang, Xin; Zhai, Chunhua; Hua, Chenlei; Qiu, Min; Hao, Yujuan; Nie, Pingping; Ye, Wenwu; Wang, Yuanchao
Zoospore chemotaxis to soybean isoflavones is essential in the early stages of infection by the oomycete pathogen Phytophthora sojae. Previously, we have identified a G-protein α subunit encoded by PsGPA1 which regulates the chemotaxis and pathogenicity of P. sojae. In the present study, we used affinity purification to identify PsGPA1-interacting proteins, including PsHint1, a histidine triad (HIT) domain-containing protein orthologous to human HIT nucleotide-binding protein 1 (HINT1). PsHint1 interacted with both the guanosine triphosphate (GTP)- and guanosine diphosphate (GDP)-bound forms of PsGPA1. An analysis of the gene-silenced transformants revealed that PsHint1 was involved in the chemotropic response of zoospores to the isoflavone daidzein. During interaction with a susceptible soybean cultivar, PsHint1-silenced transformants displayed significantly reduced infectious hyphal extension and caused a strong cell death in plants. In addition, the transformants displayed defective cyst germination, forming abnormal germ tubes that were highly branched and exhibited apical swelling. These results suggest that PsHint1 not only regulates chemotaxis by interacting with PsGPA1, but also participates in a Gα-independent pathway involved in the pathogenicity of P. sojae.
Wang, Xi; Li, Yong; Ji, Wei; Bai, Xi; Cai, Hua; Zhu, Dan; Sun, Xiao-Li; Chen, Lian-Jiang; Zhu, Yan-Ming
Tonoplast intrinsic protein (TIP) is a subfamily of the aquaporin (AQP), also known as major intrinsic protein (MIP) family, and regulates water movement across vacuolar membranes. Some reports have implied that TIP genes are associated with plant tolerance to some abiotic stresses that cause water loss, such as drought and high salinity. In our previous work, we found that an expressed sequence tag (EST) representing a TIP gene in our Glycine soja EST library was inducible by abiotic stresses. This TIP was subsequently isolated from G. soja with cDNA library screening, EST assembly and PCR, and named as GsTIP2;1. The expression patterns of GsTIP2;1 in G. soja under low temperature, salt and dehydration stress were different in leaves and roots. Though GsTIP2;1 is a stress-induced gene, overexpression of GsTIP2;1 in Arabidopsis thaliana depressed tolerance to salt and dehydration stress, but did not affect seedling growth under cold or favorable conditions. Higher dehydration speed was detected in Arabidopsis plants overexpressing GsTIP2;1, implying GsTIP2;1 might mediate stress sensitivity by enhancing water loss in the plant. Such a result is not identical to previous reports, providing some new information about the relationship between TIP and plant abiotic stress tolerance.
Dou, Daolong; Kale, Shiv D; Liu, Tingli; Tang, Qinghua; Wang, Xia; Arredondo, Felipe D; Basnayake, Shiromi; Whisson, Stephen; Drenth, Andre; Maclean, Don; Tyler, Brett M
At least 12 avirulence genes have been genetically identified and mapped in Phytophthora sojae, an oomycete pathogen causing root and stem rot of soybean. Previously, the Avr4 and Avr6 genes of P. sojae were genetically mapped within a 24 kb interval of the genome. Here, we identify Avr4 and Avr6 and show that they are actually a single gene, Avr4/6, located near the 24-kb region. Avr4/6 encodes a secreted protein of 123 amino acids with an RXLR-dEER protein translocation motif. Transient expression of Avr4/6 in soybean leaves revealed that its gene product could trigger a hypersensitive response (HR) in the presence of either Rps4 or Rps6. Silencing Avr4/6 in P. sojae stable transformants abolished the avirulence phenotype exhibited on both Rps4 and Rps6 soybean cultivars. The N terminus of Avr4/6, including the dEER motif, is sufficient to trigger Rps4-dependent HR while its C terminus is sufficient to trigger Rps6-mediated HR. Compared with alleles from avirulent races, alleles of Avr4/6 from virulent races possess nucleotide substitutions in the 5' untranslated region of the gene but not in the protein-coding region.
Andrade, Gilberto Santos; Serrão, José Eduardo; Zanuncio, José Cola; Zanuncio, Teresinha Vinha; Leite, Germano Leão Demolin; Polanczyk, Ricardo Antonio
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. PMID:20975929
Buyukkileci, Ali Oguz; Lahore, Marcello Fernandez; Tari, Canan
The production of exo-polygalacturonase (exo-PG) from orange peel (OP), a food industrial waste, using Aspergillus sojae was studied in submerged culture. A simple, low-cost, industrially significant medium formulation, composed of only OP and (NH4)2SO4 (AS) was developed. At an inoculum size of 2.8 × 10(3) spores/mL, growth was in the form of pellets, which provided better mixing of the culture broth and higher exo-PG activity. These pellets were successfully used as an inoculum for bioreactors and 173.0 U/mL exo-PG was produced. Fed-batch cultivation further enhanced the exo-PG activity to 244.0 U/mL in 127.5 h. The final morphology in the form of pellets is significant to industrial fermentation easing the subsequent downstream processing. Furthermore, the low pH trend obtained during this fermentation serves an advantage to fungal fermentations prone to contamination problems. As a result, an economical exo-PG production process was defined utilizing a food industrial by-product and producing high amount of enzyme.
Lu, Yonghai; Lam, Honming; Pi, Erxu; Zhan, Qinglei; Tsai, Sauna; Wang, Chunmei; Kwan, Yiuwa; Ngai, Saiming
Metabolomics is developing as an important functional genomics tool for understanding plant systems' response to genetic and environmental changes. Here, we characterized the metabolic changes of cultivated soybean C08 (Glycine max L. Merr) and wild soybean W05 (Glycine soja Sieb.et Zucc.) under salt stress using MS-based metabolomics, in order to reveal the phenotypes of their eight hybrid offspring (9H0086, 9H0124, 9H0391, 9H0736, 9H0380, 9H0400, 9H0434, and 9H0590). Total small molecule extracts of soybean seedling leaves were profiled by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-Fourier transform mass spectrometry (LC-FT/MS). We found that wild soybean contained higher amounts of disaccharides, sugar alcohols, and acetylated amino acids than cultivated soybean, but with lower amounts of monosaccharides, carboxylic acids, and unsaturated fatty acids. Further investigations demonstrated that the ability of soybean to tolerate salt was mainly based on synthesis of compatible solutes, induction of reactive oxygen species (ROS) scavengers, cell membrane modifications, and induction of plant hormones. On the basis of metabolic phenotype, the salt-tolerance abilities of 9H0086, 9H0124, 9H0391, 9H0736, 9H0380, 9H0400, 9H0434, and 9H0590 were discriminated. Our results demonstrated that MS-based metabolomics provides a fast and powerful approach to discriminate the salt-tolerance characteristics of soybeans.
Zhu, Dan; Bai, Xi; Zhu, Yan-Ming; Cai, Hua; Li, Yong; Ji, Wei; Chen, Chao; An, Lin; Zhu, Yi
Using homologous cloning and RT-PCR technology, we isolated a novel TIFY family gene, GsTIFY11b, from Glycine soja L. G07256, a species that is tolerant to saline and alkaline environments. Phylogenetic analysis indicated that GsTIFY11b was closely related to AtTIFY11a with 56% similarity in amino acid identity. Protein sequence analysis showed that GsTIFY11b protein also had conserved TIFY domain, N-terminal domain, and a C-terminal Jas motif. Quantitative realtime PCR analysis indicated that the expression of GsTIFY11b was induced by both saline and alkaline stresses. Two homozygous GsTIFY11b over-expressing transgenic Arabidopsis lines were obtained. Phenotypic analysis of the transgenic and wild-type Arabidopsis indicated that over-expressing GsTIFY11b in Arabidopsis did not enhance plant tolerance to saline and alkaline stresses, whereas it showed an increased sensitivity to saline stress during seed germination and seedling development. Expression analysis of saline stress response marker genes in transgenic and wild-type plants under stress condition indicated that GsTIFY11b regulated the expression of RD29B, KIN1, and DREB. The transient expression of a GsTIFY11b-GFP fusion protein in onion epidermal cells showed that GsTIFY11b was localized to the nucleus, suggesting a role as a transcriptional regulator in the saline stress response pathway.
Arnemann, J A; Tay, W T; Walsh, T K; Brier, H; Gordon, K; Hickmann, F; Ugalde, G; Guedes, J V C
Soybean Stem Fly (SSF), Melanagromyza sojae (Zehntner), belongs to the family Agromyzidae and is highly polyphagous, attacking many plant species of the family Fabaceae, including soybean and other beans. SSF is regarded as one of the most important pests in soybean fields of Asia (e.g., China, India), North East Africa (e.g., Egypt), parts of Russia, and South East Asia. Despite reports of Agromyzidae flies infesting soybean fields in Rio Grande do Sul State (Brazil) in 1983 and 2009 and periodic interceptions of SSF since the 1940s by the USA quarantine authorities, SSF has not been officially reported to have successfully established in the North and South Americas. In South America, M. sojae was recently confirmed using morphology and its complete mitochondrial DNA (mtDNA) was characterized. In the present study, we surveyed the genetic diversity of M. sojae, collected directly from soybean host plants, using partial mtDNA cytochrome oxidase I (COI) gene, and provide evidence of multiple (>10) maternal lineages in SSF populations in South America, potentially representing multiple incursion events. However, a single incursion involving multiple-female founders could not be ruled out. We identified a haplotype that was common in the fields of two Brazilian states and the individuals collected from Australia in 2013. The implications of SSF incursions in southern Brazil are discussed in relation to the current soybean agricultural practices, highlighting an urgent need for better understanding of SSF population movements in the New World, which is necessary for developing effective management options for this significant soybean pest.
Tuyen, D D; Lal, S K; Xu, D H
Salt-affected soils are generally classified into two main categories, sodic (alkaline) and saline. Our previous studies showed that the wild soybean accession JWS156-1 (Glycine soja) from the Kinki area of Japan was tolerant to NaCl salt, and the quantitative trait locus (QTL) for NaCl salt tolerance was located on soybean linkage group N (chromosome 3). Further investigation revealed that the wild soybean accession JWS156-1 also had a higher tolerance to alkaline salt stress. In the present study, an F(6) recombinant inbred line mapping population (n = 112) and an F(2) population (n = 149) derived from crosses between a cultivated soybean cultivar Jackson and JWS156-1 were used to identify QTL for alkaline salt tolerance in soybean. Evaluation of soybean alkaline salt tolerance was carried out based on salt tolerance rating (STR) and leaf chlorophyll content (SPAD value) after treatment with 180 mM NaHCO(3) for about 3 weeks under greenhouse conditions. In both populations, a significant QTL for alkaline salt tolerance was detected on the molecular linkage group D2 (chromosome 17), which accounted for 50.2 and 13.0% of the total variation for STR in the F(6) and the F(2) populations, respectively. The wild soybean contributed to the tolerance allele in the progenies. Our results suggest that QTL for alkaline salt tolerance is different from the QTL for NaCl salt tolerance found previously in this wild soybean genotype. The DNA markers closely associated with the QTLs might be useful for marker-assisted selection to pyramid tolerance genes in soybean for both alkaline and saline stresses.
Wang, Xi; Cai, Hua; Li, Yong; Zhu, Yanming; Ji, Wei; Bai, Xi; Zhu, Dan; Sun, Xiaoli
Plasma membrane intrinsic proteins (PIPs) belong to the aquaporin family and facilitate water movement across plasma membranes. Existing data indicate that PIP genes are associated with the abilities of plants to tolerate certain stress conditions. A review of our Glycine soja expressed sequence tag (EST) dataset revealed that abiotic stress stimulated expression of a PIP, herein designated as GsPIP2;1 (GenBank_Accn: FJ825766). To understand the roles of this PIP in stress tolerance, we generated a coding sequence for GsPIP2;1 by in silico elongation and cloned the cDNA by 5'-RACE. Semiquantitative RT-PCR showed that GsPIP2;1 expression was stimulated in G. soja leaves by cold, salt, or dehydration stress, whereas the same stresses suppressed GsPIP2;1 expression in the roots. Transgenic Arabidopsis thaliana plants overexpressing GsPIP2;1 grew normally under unstressed and cold conditions, but exhibited depressed tolerance to salt and dehydration stresses. Moreover, greater changes in water potential were detected in the transgenic A. thaliana shoots, implying that GsPIP2;1 may negatively impact stress tolerance by regulating water potential. These results, deviating from those obtained in previous reports, provide new insights into the relationship between PIPs and abiotic stress tolerance in plants.
Wang, Qinhu; Li, Tingting; Xu, Ke; Zhang, Wei; Wang, Xiaolong; Quan, Junli; Jin, Weibo; Zhang, Meixiang; Fan, Guangjin; Wang, Ming-Bo; Shan, Weixing
Along with the well-studied microRNA (miRNA) and small interfering RNA (siRNA) is a new class of transfer RNA-derived small RNA (tsRNA), which has recently been detected in multiple organisms and is implicated in gene regulation. However, while miRNAs and siRNAs are known to repress gene expression through sequence-specific RNA cleavage or translational repression, how tsRNAs regulate gene expression remains unclear. Here we report the identification and functional characterization of tsRNAs in the oomycete pathogen Phytophthora sojae. We show that multiple tRNAs are processed into abundant tsRNAs, which accumulate in a similar developmental stage-specific manner and are negatively correlated with the expression of predicted target genes. Degradome sequencing and 5′ RLM RACE experiments indicate tsRNAs can trigger degradation of target transcripts. Transient expression assays using GUS sensor constructs confirmed the requirement of sequence complementarity in tsRNA-mediated RNA degradation in P. sojae. Our results show that the tsRNA are a class of functional endogenous sRNAs and suggest that tsRNA regulate gene expression through inducing sequence-specific degradation of target RNAs in oomycetes. PMID:28066490
Oshima, Hisaka; Kimura, Isao; Izumori, Ken
Xylosylpsicoses (XPs) were synthesized by transglycosylation (TG) with arabinoxylan (AX) and D-psicose (D-psi) using endo-1,4-beta-D-xylanase (EC 220.127.116.11) from Aspergillus sojae. The structures of XPs were analyzed by HPLC to determine sugar composition and molecular mass, by methylation analysis using GC-MS to determine linkage, and by 1H- and 13C-NMR spectrometries to obtain the anomeric configuration of glycosidic linkage. By chemical analysis, it was found that the structures of XPs are Xylp beta1-4Psip, Xylp beta1-5Psip, Xylp beta1-1Psif and Xylp beta1-6Psif.
Chen, Peng; Yan, Kun; Shao, Hongbo; Zhao, Shijie
Glycine soja (BB52) is a wild soybean cultivar grown in coastal saline land in Yellow River Delta, China. In order to reveal the physiological mechanisms adapting to salinity, we examined photosynthesis, ion flux, antioxidant system and water status in Glycine soja under NaCl treatments, taking a cultivated soybean, ZH13, as control. Upon NaCl exposure, higher relative water content and water potential were maintained in the leaf of BB52 than ZH13, which might depend on the more accumulation of osmotic substances such as glycinebetaine and proline. Compared with ZH13, activities of antioxidant enzymes including superoxide dismutase, catalase, ascorbate peroxidase and contents of ascorbate, glutathione and phenolics were enhanced to a higher level in BB52 leaf under NaCl stress, which could mitigate the salt-induced oxidative damage in BB52. Consistently, lipid peroxidation indicated by malondialdehyde content was lower in BB52 leaf. Photosynthetic rate (Pn) was decreased by NaCl stress in BB52 and ZH13, and the decrease was greater in ZH13. The decreased Pn in BB52 was mainly due to stomatal limitation. The inhibited activation of rubisco enzyme in ZH13 due to the decrease of rubisco activase content became an important limiting factor of Pn, when NaCl concentration increased to 200 mM. Rubisco activase in BB52 was not affected by NaCl stress. Less negative impact in BB52 derived from lower contents of Na(+) and Cl(-) in the tissues, and non-invasive micro-test technique revealed that BB52 roots had higher ability to extrude Na(+) and Cl(-). Wild soybean is a valuable genetic resource, and our study may provide a reference for molecular biologist to improve the salt tolerance of cultivated soybean in face of farmland salinity.
Subramanian, Senthil; Graham, Madge Y.; Yu, Oliver; Graham, Terrence L.
Isoflavones are thought to play diverse roles in plant-microbe interactions and are also potentially important to human nutrition and medicine. Isoflavone synthase (IFS) is a key enzyme for the formation of the isoflavones. Here, we examined the consequences of RNAi silencing of genes for this enzyme in soybean (Glycine max). Soybean cotyledon tissues were transformed with Agrobacterium rhizogenes carrying an RNAi silencing construct designed to silence expression of both copies of IFS genes. Approximately 50% of emerging roots were transformed with the RNAi construct, and most transformed roots exhibited >95% silencing of isoflavone accumulation. Silencing of IFS was also demonstrated throughout the entire cotyledon (in tissues distal to the transformation site) both by high-performance liquid chromatography analysis of isoflavones and by real-time reverse transcription-PCR. This distal silencing led to a nearly complete suppression of mRNA accumulation for both the IFS1 and IFS2 genes and of isoflavone accumulations induced by wounding or treatment with the cell wall glucan elicitor from Phytophthora sojae. Preformed isoflavone conjugates were not reduced in distal tissues, suggesting little turnover of these stored isoflavone pools. Distal silencing was established within just 5 d of transformation and was highly efficient for a 3- to 4-d period, after which it was no longer apparent in most experiments. Silencing of IFS was effective in at least two genotypes and led to enhanced susceptibility to P. sojae, disrupting both R gene-mediated resistance in roots and nonrace-specific resistance in cotyledon tissues. The soybean cotyledon system, already a model system for defense signal-response and cell-to-cell signaling, may provide a convenient and effective system for functional analysis of plant genes through gene silencing. PMID:15778457
Sun, Xiaoli; Yang, Shanshan; Sun, Mingzhe; Wang, Sunting; Ding, Xiaodong; Zhu, Dan; Ji, Wei; Cai, Hua; Zhao, Chaoyue; Wang, Xuedong; Zhu, Yanming
It has been well demonstrated that cystatins regulated plant stress tolerance through inhibiting the cysteine proteinase activity under environmental stress. However, there was limited information about the role of cystatins in plant alkali stress response, especially in wild soybean. Here, in this study, we focused on the biological characterization of a novel Glycine soja cystatin protein GsCPI14, which interacted with the calcium/calmodulin-binding receptor-like kinase GsCBRLK and positively regulated plant alkali stress tolerance. The protein-protein interaction between GsCBRLK and GsCPI14 was confirmed by using split-ubiquitin based membrane yeast two-hybrid analysis and bimolecular fluorescence complementation assay. Expression of GsCPI14 was greatly induced by salt, ABA and alkali stress in G. soja, and GsCBRLK overexpression (OX) in Glycine max promoted the stress induction of GmCPI14 expression under stress conditions. Furthermore, we found that GsCPI14-eGFP fusion protein localized in the entire Arabidopsis protoplast and onion epidermal cell, and GsCPI14 showed ubiquitous expression in different tissues of G. soja. In addition, we gave evidence that the GST-GsCPI14 fusion protein inhibited the proteolytic activity of papain in vitro. At last, we demonstrated that OX of GsCPI14 in Arabidopsis promoted the seed germination under alkali stress, as evidenced by higher germination rates. GsCPI14 transgenic Arabidopsis seedlings also displayed better growth performance and physiological index under alkali stress. Taken together, results presented in this study demonstrated that the G. soja cysteine proteinase inhibitor GsCPI14 interacted with the calcium/calmodulin-binding receptor-like kinase GsCBRLK and regulated plant tolerance to alkali stress.
Silva, Farley W S; Elliot, Simon L
Temperature and crowding are key environmental factors mediating the transmission and epizooty of infectious disease in ectotherm animals. The host physiology may be altered in a temperature-dependent manner and thus affects the pathogen development and course of diseases within an individual and host population, or the transmission rates (or infectivity) of pathogens shift linearly with the host population density. To our understanding, the knowledge of interactive and synergistic effects of temperature and population density on the host-pathogen system is limited. Here, we tested the interactional effects of these environmental factors on phenotypic plasticity, immune defenses, and disease resistance in the velvetbean caterpillar Anticarsia gemmatalis. Upon egg hatching, caterpillars were reared in thermostat-controlled chambers in a 2 × 4 factorial design: density (1 or 8 caterpillars/pot) and temperature (20, 24, 28, or 32°C). Of the immune defenses assessed, encapsulation response was directly affected by none of the environmental factors; capsule melanization increased with temperature in both lone- and group-reared caterpillars, although the lone-reared ones presented the most evident response, and hemocyte numbers decreased with temperature regardless of the population density. Temperature, but not population density, affected considerably the time from inoculation to death of velvetbean caterpillar. Thus, velvetbean caterpillars succumbed to Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) more quickly at higher temperatures than at lower temperatures. As hypothesized, temperature likely affected caterpillars' movement rates, and thus the contact between conspecifics, which in turn affected the phenotypic expression of group-reared caterpillars. Our results suggest that environmental factors, mainly temperature, strongly affect both the course of disease in velvetbean caterpillar population and its defenses against pathogens. As a soybean
Di Bello, M M; Souza, B H S; Nogueira, L; Ribeiro, Z A; Eduardo, W I; Boiça Júnior, A L
Advances in techniques for rearing insects on artificial diets are fundamental to solving issues of basic and applied entomology. In this study, we evaluated the development of Spodoptera albula (Walker) (Lepidoptera: Noctuidae) on three artificial diets used for other species of Lepidoptera, at three larval densities, and two densities of adult couples housed in oviposition cages of two sizes, with the aim of optimizing methodology for rearing S. albula in the laboratory. Biological parameters were recorded from S. albula, and a fitness index was calculated based on the larval survival and duration and weight of pupae. The total and daily oviposition was recorded using 5 or 10 adult couples of S. albula housed in two cage sizes. Concentrations of total nitrogen and protein in the tested diets were determined. Development of S. albula was completed in all artificial diets; however, the diet used for rearing Anticarsia gemmatalis (Hübner) larvae was the most suitable for S. albula, yielding intermediate development time and higher survival relative to the other diets. Individualization of larvae favored S. albula development by producing overall greater weights of larvae and pupae, higher survival rates, and longer adult longevity. Cage size and number of couples per cage did not influence S. albula fecundity in the experiment conditions. Spodoptera albula can be satisfactorily reared on the artificial diet used for A. gemmatalis, using one larva per tube, and either density of adults at any cage size. Additional amendments are needed in the rearing methodology to achieve optimal conditions for larval development to adulthood.
Huang, Wei-Qian; Zhong, Li-Fen; Meng, Zhi-Zhong; You, Zi-Juan; Li, Jia-Zhou; Luo, Xiao-Chun
A leucine aminopeptidase Lap1 was cloned from Aspergillus sojae GIM3.30. The truncated Lap1 without a signal peptide was over-expressed in P. pastoris, and the enzymatic characteristics of recombinant Lap1 (rLap1) were tested. The rLap1 was about 36.7 kDa with an optimal pH 8.0 and optimal temperature 50 °C for substrate Leu-p-nitroanilide and it sustained 50 % activity after 1 h incubation at 50 °C. The activity of rLap1 was significantly inhibited by EDTA, whereas Co(2+), Mn(2+), and Ca(2+) ions, but not Zn(2+) ions, restored its activity. rLap1 showed the highest activity against Arg-pNA and then Leu-, Lys-, Met-, and Phe-pNA. The 3D structure of rLap1 showed it had a conserved functional charge/dipole complex and a hydrogen bond network of Zn2-D179-S228-Q177-D229-S158 around its active center. An acidic Asp residue was found at the bottom of the substrate binding pocket, which explains its preference for basic N-terminal amino acid substrates such as Arg and Lys. rLap1 improved the degree of hydrolysis of casein and soy protein hydrolysates and also decreased their bitterness, indicating its potential utility in food production.
He, Shuilian; Wang, Yunsheng; Volis, Sergei; Li, Dezhu; Yi, Tingshuang
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.
Mizuguti, Aki; Ohigashi, Kentaro; Yoshimura, Yasuyuki; Kaga, Akito; Kuroda, Yosuke; Matsuo, Kazuhito
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.
Liu, Ailin; Yu, Yang; Duan, Xiangbo; Sun, Xiaoli; Duanmu, Huizi; Zhu, Yanming
Plant SKP1-like family proteins, components of the SCF complex E3 ligases, are involved in the regulation of plant development and stress responses. Little is known about the precise function of SKP genes in plant responses to environmental stresses. GsSKP21 was initially identified as a potential stress-responsive gene based on the transcriptome sequencing of Glycine soja. In this study, we found that GsSKP21 protein contains highly conserved SKP domains in its N terminus and an extra unidentified domain in its C terminus. The transcript abundance of GsSKP21, detected by quantitative real-time PCR, was induced under the treatment of alkali and salt stresses. Overexpression of GsSKP21 in Arabidopsis dramatically increased plant tolerance to alkali stress. Furthermore, we found that overexpression of GsSKP21 resulted in decreased ABA sensitivity during both the seed germination and early seedling growth stages. GsSKP21 mediated ABA signaling by altering the expression levels of the ABA signaling-related and ABA-induced genes. We also investigated the tissue expression specificity and subcellular localization of GsSKP21. These results suggest that GsSKP21 is important for plant tolerance to alkali stress and plays a critical regulatory role in the ABA-mediated stress response.
Li, Qi; Zhang, Meixiang; Shen, Danyu; Liu, Tingli; Chen, Yanyu; Zhou, Jian-Min; Dou, Daolong
Oomycete pathogens produce a large number of effectors to promote infection. Their mode of action are largely unknown. Here we show that a Phytophthora sojae effector, PsCRN63, suppresses flg22-induced expression of FRK1 gene, a molecular marker in pathogen-associated molecular patterns (PAMP)-triggered immunity (PTI). However, PsCRN63 does not suppress upstream signaling events including flg22-induced MAPK activation and BIK1 phosphorylation, indicating that it acts downstream of MAPK cascades. The PsCRN63-transgenic Arabidopsis plants showed increased susceptibility to bacterial pathogen Pseudomonas syringae pathovar tomato (Pst) DC3000 and oomycete pathogen Phytophthora capsici. The callose deposition were suppressed in PsCRN63-transgenic plants compared with the wild-type control plants. Genes involved in PTI were also down-regulated in PsCRN63-transgenic plants. Interestingly, we found that PsCRN63 forms an dimer that is mediated by inter-molecular interactions between N-terminal and C-terminal domains in an inverted association manner. Furthermore, the N-terminal and C-terminal domains required for the dimerization are widely conserved among CRN effectors, suggesting that homo-/hetero-dimerization of Phytophthora CRN effectors is required to exert biological functions. Indeed, the dimerization was required for PTI suppression and cell death-induction activities of PsCRN63. PMID:27243217
Chen, Chen; Sun, Xiaoli; Duanmu, Huizi; Yu, Yang; Liu, Ailin; Xiao, Jialei; Zhu, Yanming
Myo-inositol participates in various aspects of plant physiology, and myo-inositol oxygenase is the key enzyme of the myo-inositol oxygenation pathway. Previous studies indicated that myo-inositol oxygenase may play a role in plant responses to abiotic stresses. In this study, we focused on the functional characterization of GsMIOX1a, a remarkable alkaline stress-responsive gene of Glycine soja 07256, based on RNA-seq data. Using quantitative real-time PCR, we demonstrated that GsMIOX1a is rapidly induced by alkaline stress and expressed predominantly in flowers. We also elucidated the positive function of GsMIOX1a in the alkaline response in the wild type, atmiox1 mutant as well as GsMIOX1a-overexpressing Arabidopsis. We determined that atmiox1 mutant decreased Arabidopsis tolerance to alkaline stress, whereas GsMIOX1a overexpression increased tolerance. Moreover, the expression levels of some alkaline stress-responsive and inducible marker genes, including H+-Ppase, NADP-ME, KIN1 and RD29B, were also up-regulated in GsMIOX1a overexpression lines compared with the wild type and atmiox1 mutant. Together, these results suggest that the GsMIOX1a gene positively regulates plant tolerance to alkaline stress. This is the first report to demonstrate that ectopic expression of myo-inositol oxygenase improves alkaline tolerance in plants. PMID:26091094
Zhang, Hengyou; Li, Chunying; Davis, Eric L.; Wang, Jinshe; Griffin, Joshua D.; Kofsky, Janice; Song, Bao-Hua
Soybean cyst nematode (SCN) is the most destructive soybean pest worldwide. Host plant resistance is the most environmentally friendly and cost-effective way of mitigating SCN damage to soybeans. However, overuse of the resistant soybean [Glycine max (L.) Merr.] cultivars from limited genetic resources has resulted in SCN race shifts in many soybean-growing areas. Thus, exploration of novel sources of SCN resistance and dissection of the genetic basis are urgently needed. In this study, we screened 235 wild soybean (Glycine soja Sieb. & Zucc.) accessions to identify genotypes resistant to SCN HG Type 2.5.7 (race 5), a less investigated type but is prevalent in the southeastern US. We also dissected the genetic basis of SCN resistance using a genome-wide association study with SNPs genotyped by SoySNP50k iSelect BeadChip. In total, 43 resistant accessions (female index < 30) were identified, with 10 SNPs being significantly associated with SCN HG 2.5.7 resistance in this wild species. Furthermore, four significant SNPs were localized to linked regions of the known quantitative trait locus (QTL) rhg1 on chromosome 18. The other four SNPs on chromosome 18 and two SNPs on chromosome 19 are novel. Genes encoding disease resistance-related proteins with a leucine-rich region, a mitogen-activated protein kinase (MAPK) on chromosome 18, and a MYB transcription factor on chromosome 19 were identified as promising candidate genes. The identified SNPs and candidate genes will benefit future marker-assisted breeding and dissection of the molecular mechanisms underlying the soybean-SCN interaction. PMID:27582748
Goto, Hidetoshi; Shimada, Hiroshi; Horak, Michael J.; Ahmad, Aqeel; Baltazar, Baltazar M.; Perez, Tim; McPherson, Marc A.; Stojšin, Duška; Shimono, Ayako; Ohsawa, Ryo
Insect-protected soybean (Glycine max (L.) Merr.) was developed to protect against foliage feeding by certain Lepidopteran insects. The assessment of potential consequences of transgene introgression from soybean to wild soybean (Glycine soja Seib. et Zucc.) is required as one aspect of the environmental risk assessment (ERA) in Japan. A potential hazard of insect-protected soybean may be hypothesized as transfer of a trait by gene flow to wild soybean and subsequent reduction in foliage feeding by Lepidopteran insects that result in increased weediness of wild soybean in Japan. To assess this potential hazard two studies were conducted. A three-year survey of wild soybean populations in Japan was conducted to establish basic information on foliage damage caused by different herbivores. When assessed across all populations and years within each prefecture, the total foliage from different herbivores was ≤ 30%, with the lowest levels of defoliation (< 2%) caused by Lepidopteran insects. A separate experiment using five levels of simulated defoliation (0%, 10%, 25%, 50% and 100%) was conducted to assess the impact on pod and seed production and time to maturity of wild soybean. The results indicated that there was no decrease in wild soybean plants pod or seed number or time to maturity at defoliation rates up to 50%. The results from these experiments indicate that wild soybean is not limited by lepidopteran feeding and has an ability to compensate for defoliation levels observed in nature. Therefore, the potential hazard to wild soybean from the importation of insect-protected soybean for food and feed into Japan is negligible. PMID:26963815
Reiss, Kerstin; Kirchner, Eva; Gijzen, Mark; Zocher, Georg; Löffelhardt, Birgit; Nürnberger, Thorsten; Stehle, Thilo; Brunner, Frédéric
Transglutaminases (TGases) are ubiquitous enzymes that catalyze selective cross-linking between protein-bound glutamine and lysine residues; the resulting isopeptide bond confers high resistance to proteolysis. Phytophthora sojae, a pathogen of soybean, secretes a Ca(2+)-dependent TGase (GP42) that is activating defense responses in both host and non-host plants. A GP42 fragment of 13 amino acids, termed Pep-13, was shown to be absolutely indispensable for both TGase and elicitor activity. GP42 does not share significant primary sequence similarity with known TGases from mammals or bacteria. This suggests that GP42 has evolved novel structural and catalytic features to support enzymatic activity. We have solved the crystal structure of the catalytically inactive point mutant GP42 (C290S) at 2.95 Å resolution and identified residues involved in catalysis by mutational analysis. The protein comprises three domains that assemble into an elongated structure. Although GP42 has no structural homolog, its core region displays significant similarity to the catalytic core of the Mac-1 cysteine protease from Group A Streptococcus, a member of the papain-like superfamily of cysteine proteases. Proteins that are taxonomically related to GP42 are only present in plant pathogenic oomycetes belonging to the order of the Peronosporales (e.g. Phytophthora, Hyaloperonospora, and Pythium spp.) and in marine Vibrio bacteria. This suggests that a lateral gene transfer event may have occurred between bacteria and oomycetes. Our results offer a basis to design and use highly specific inhibitors of the GP42-like TGase family that may impair the growth of important oomycete and bacterial pathogens.
Goto, Hidetoshi; Shimada, Hiroshi; Horak, Michael J; Ahmad, Aqeel; Baltazar, Baltazar M; Perez, Tim; McPherson, Marc A; Stojšin, Duška; Shimono, Ayako; Ohsawa, Ryo
Insect-protected soybean (Glycine max (L.) Merr.) was developed to protect against foliage feeding by certain Lepidopteran insects. The assessment of potential consequences of transgene introgression from soybean to wild soybean (Glycine soja Seib. et Zucc.) is required as one aspect of the environmental risk assessment (ERA) in Japan. A potential hazard of insect-protected soybean may be hypothesized as transfer of a trait by gene flow to wild soybean and subsequent reduction in foliage feeding by Lepidopteran insects that result in increased weediness of wild soybean in Japan. To assess this potential hazard two studies were conducted. A three-year survey of wild soybean populations in Japan was conducted to establish basic information on foliage damage caused by different herbivores. When assessed across all populations and years within each prefecture, the total foliage from different herbivores was ≤ 30%, with the lowest levels of defoliation (< 2%) caused by Lepidopteran insects. A separate experiment using five levels of simulated defoliation (0%, 10%, 25%, 50% and 100%) was conducted to assess the impact on pod and seed production and time to maturity of wild soybean. The results indicated that there was no decrease in wild soybean plants pod or seed number or time to maturity at defoliation rates up to 50%. The results from these experiments indicate that wild soybean is not limited by lepidopteran feeding and has an ability to compensate for defoliation levels observed in nature. Therefore, the potential hazard to wild soybean from the importation of insect-protected soybean for food and feed into Japan is negligible.
Sun, XiaoLi; Sun, Mingzhe; Luo, Xiao; Ding, XiaoDong; Ji, Wei; Cai, Hua; Bai, Xi; Liu, XiaoFei; Zhu, YanMing
Receptor such as protein kinases are proposed to work as sensors to initiate signaling cascades in higher plants. However, little is known about the precise functions of receptor such as protein kinases in abiotic stress response in plants, especially in wild soybean. Here, we focused on characterization of the biological functions of a receptor-like cytoplasmic serine/threonine protein kinase gene, GsRLCK, which was previously identified as a putative salt-alkali stress-related gene from the transcriptome profiles of Glycine soja. Bioinformatic analysis showed that GsRLCK protein contained a conserved kinase catalytic domain and two transmembrane domains at the N-terminus, but no typical extracellular domain. Consistently, GsRLCK-eGFP fusion protein was observed on the plasma membrane, but eGFP alone was distributing throughout the cytoplasm in onion epidermal cells. Quantitative real-time PCR analysis revealed the induced expression of GsRLCK by ABA, salt, alkali, and drought stresses. However, the expression levels of GsRLCK seemed to be similar in different tissues, except soybean pod. Phenotypic assays demonstrated that GsRLCK overexpression decreased ABA sensitivity and altered expression levels of ABA-responsive genes. Furthermore, we also found that GsRLCK conferred increased tolerance to salt and drought stresses and increased expression levels of a handful of stress-responsive genes, when overexpressing in Arabidopsis. In a word, we gave exact evidence that GsRLCK was a novel receptor-like cytoplasmic protein kinase and played a crucial role in plant responses to ABA, salt, and drought stresses.
Xin, Dawei; Qi, Zhaoming; Jiang, Hongwei; Hu, Zhenbang; Zhu, Rongsheng; Hu, Jiahui; Han, Heyu; Hu, Guohua; Liu, Chunyan; Chen, Qingshan
Increasing the yield of soybean (Glycine max L. Merrill) is a main aim of soybean breeding. The 100-seed weight is a critical factor for soybean yield. To facilitate genetic analysis of quantitative traits and to improve the accuracy of marker-assisted breeding in soybean, a valuable mapping population consisting of 194 chromosome segment substitution lines (CSSLs) was developed. In these lines, different chromosomal segments of the Chinese cultivar Suinong 14 were substituted into the genetic background of wild soybean (Glycine soja Sieb. & Zucc.) ZYD00006. Based on these CSSLs, a genetic map covering the full genome was generated using 121 simple sequence repeat (SSR) markers. In the quantitative trait loci (QTL) analysis, twelve main effect QTLs (qSW-B1-1/2/3, qSW-D1b-1/2, qSW-D2-1/2, qSW-G-1/2/3, qSW-M-2 and qSW-N-2) underlying 100-seed weight were identified in 2011 and 2012. The epistatic effects of pairwise interactions between markers were analyzed in 2011 and 2012. The results clearly demonstrated that these CSSLs could be used to identify QTLs, and that an epistatic analysis was able to detect several sites with important epistatic effects on 100-seed weight. Thus, we identified loci that will be valuable for improving soybean 100-seed weight. These results provide a valuable foundation for identifying the precise location of genes of interest, and for designing cloning and marker-assisted selection breeding strategies targeting the 100-seed weight of soybean.
Sun, Xiaoli; Luo, Xiao; Sun, Mingzhe; Chen, Chao; Ding, Xiaodong; Wang, Xuedong; Yang, Shanshan; Yu, Qingyue; Jia, Bowei; Ji, Wei; Cai, Hua; Zhu, Yanming
It is well established that 14-3-3 proteins are key regulators of multiple stress signal transduction cascades. However, the biological functions of soybean 14-3-3 proteins, especially in plant drought response, are not yet known. In this study, we characterized a Glycine soja 14-3-3 gene, GsGF14o, which is involved in plant development and drought response. GsGF14o expression was greatly induced by drought stress, as evidenced by the quantitative real-time PCR and β-glucuronidase (GUS) activity analysis. GsGF14o overexpression in Arabidopsis thaliana resulted in decreased drought tolerance during seed germination and seedling growth. Furthermore, silencing of AtGF14µ, the most homologous 14-3-3 gene of GsGF14o, led to enhanced drought tolerance at both the seed germination and seedling stage. Unexpectedly, GsGF14o transgenic lines showed reduced water loss and transpiration rates compared with wild-type plants, which was demonstrated to be the consequence of the decreased stomatal size. At the same time, the smaller stomata due to GsGF14o overexpression led to a relatively slow net photosynthesis rate, which led to a growth penalty under drought stress. We further demonstrated that GsGF14o overexpression caused deficits in root hair formation and development, and thereby reduced the water intake capacity of the transgenic root system. In addition, GsGF14o overexpression down-regulated the transcript levels of drought-responsive marker genes. Finally, we also investigated the tissue-specific accumulation of GsGF14o by using a GUS activity assay. Collectively, the results presented here confirm that GsGF14o plays a dual role in drought stress responses through its involvement in the regulation of stomatal size and root hair development.
Zhang, Hengyou; Li, Chunying; Davis, Eric L; Wang, Jinshe; Griffin, Joshua D; Kofsky, Janice; Song, Bao-Hua
Soybean cyst nematode (SCN) is the most destructive soybean pest worldwide. Host plant resistance is the most environmentally friendly and cost-effective way of mitigating SCN damage to soybeans. However, overuse of the resistant soybean [Glycine max (L.) Merr.] cultivars from limited genetic resources has resulted in SCN race shifts in many soybean-growing areas. Thus, exploration of novel sources of SCN resistance and dissection of the genetic basis are urgently needed. In this study, we screened 235 wild soybean (Glycine soja Sieb. & Zucc.) accessions to identify genotypes resistant to SCN HG Type 2.5.7 (race 5), a less investigated type but is prevalent in the southeastern US. We also dissected the genetic basis of SCN resistance using a genome-wide association study with SNPs genotyped by SoySNP50k iSelect BeadChip. In total, 43 resistant accessions (female index < 30) were identified, with 10 SNPs being significantly associated with SCN HG 2.5.7 resistance in this wild species. Furthermore, four significant SNPs were localized to linked regions of the known quantitative trait locus (QTL) rhg1 on chromosome 18. The other four SNPs on chromosome 18 and two SNPs on chromosome 19 are novel. Genes encoding disease resistance-related proteins with a leucine-rich region, a mitogen-activated protein kinase (MAPK) on chromosome 18, and a MYB transcription factor on chromosome 19 were identified as promising candidate genes. The identified SNPs and candidate genes will benefit future marker-assisted breeding and dissection of the molecular mechanisms underlying the soybean-SCN interaction.
Yang, Liang; Ji, Wei; Gao, Peng; Li, Yong; Cai, Hua; Bai, Xi; Chen, Qin; Zhu, Yanming
Plant Snf1 (sucrose non-fermenting-1) related protein kinase (SnRK), a subfamily of serine/threonine kinases, has been implicated as a crucial upstream regulator of ABA and osmotic signaling as in many other signaling cascades. In this paper, we have isolated a novel plant specific ABA activated calcium independent protein kinase (GsAPK) from a highly salt tolerant plant, Glycine soja (50109), which is a member of the SnRK2 family. Subcellular localization studies using GFP fusion protein indicated that GsAPK is localized in the plasma membrane. We found that autophosphorylation and Myelin Basis Protein phosphorylation activity of GsAPK is only activated by ABA and the kinase activity also was observed when calcium was replaced by EGTA, suggesting its independence of calcium in enzyme activity. We also found that cold, salinity, drought, and ABA stress alter GsAPK gene transcripts and heterogonous overexpression of GsAPK in Arabidopsis alters plant tolerance to high salinity and ABA stress. In summary, we demonstrated that GsAPK is a Glycine soja ABA activated calcium independent SnRK-type kinase presumably involved in ABA mediated stress signal transduction.
In vitro translocation experiments with RxLR-reporter fusion proteins of Avr1b from Phytophthora sojae and AVR3a from Phytophthora infestans fail to demonstrate specific autonomous uptake in plant and animal cells.
Wawra, Stephan; Djamei, Armin; Albert, Isabell; Nürnberger, Thorsten; Kahmann, Regine; van West, Pieter
Plant-pathogenic oomycetes have a large set of secreted effectors that can be translocated into their host cells during infection. One group of these effectors are the RxLR effectors for which it has been shown, in a few cases, that the RxLR motif is important for their translocation. It has been suggested that the RxLR-leader sequences alone are enough to translocate the respective effectors into eukaryotic cells through binding to surface-exposed phosphoinositol-3-phosphate. These conclusions were primary based on translocation experiments conducted with recombinant fusion proteins whereby the RxLR leader of RxLR effectors (i.e., Avr1b from Phytophthora sojae) were fused to the green fluorescent protein reporter-protein. However, we failed to observe specific cellular uptake for a comparable fusion protein where the RxLR leader of the P. infestans AVR3a was fused to monomeric red fluorescent protein. Therefore, we reexamined the ability of the reported P. sojae AVR1b RxLR leader to enter eukaryotic cells. Different relevant experiments were performed in three independent laboratories, using fluorescent reporter fusion constructs of AVR3a and Avr1b proteins in a side-by-side comparative study on plant tissue and human and animal cells. We report that we were unable to obtain conclusive evidence for specific RxLR-mediated translocation.
Guo, Yong; Qiu, Li-Juan
Color is one of the phenotypic markers mostly used to study soybean (Glycine max L. Merr.) genetic, molecular and biochemical processes. Two P450-dependent mono-oxygenases, flavonoid 3'-hydroxylase (F3'H; EC18.104.22.168) and flavonoid 3',5'-hydroxylase (F3'5'H, EC22.214.171.124), both catalyzing the hydroxylation of the B-ring in flavonoids, play an important role in coloration. Previous studies showed that the T locus was a gene encoding F3'H and the W1 locus co-segregated with a gene encoding F3'5'H in soybean. These two genetic loci have identified to control seed coat, flower and pubescence colors. However, the allelic distributions of both F3'H and F3'5'H genes in soybean were unknown. In this study, three novel alleles were identified (two of four alleles for GmF3'H and one of three alleles for GmF3'5'H). A set of gene-tagged markers was developed and verified based on the sequence diversity of all seven alleles. Furthermore, the markers were used to analyze soybean accessions including 170 cultivated soybeans (G. max) from a mini core collection and 102 wild soybeans (G. soja). For both F3'H and F3'5'H, the marker selection efficiencies for pubescence color and flower color were determined. The results showed that one GmF3'H allele explained 92.2 % of the variation in tawny and two gmf3'h alleles explained 63.8 % of the variation in gray pubescence colors. In addition, two GmF3'5'H alleles and one gmF3'5'h allele explained 94.0 % of the variation in purple and 75.3 % in white flowers, respectively. By the combination of the two loci, seed coat color was determined. In total, 90.9 % of accessions possessing both the gmf3'h-b and gmf3'5'h alleles had yellow seed coats. Therefore, seed coat colors are controlled by more than two loci.
Zhang, Jinlong; Wang, Jiaxue; Jiang, Wei; Liu, Juge; Yang, Songnan; Gai, Junyi; Li, Yan
Soil alkalinity is a major abiotic constraint to crop productivity and quality. Wild soybean (Glycine soja) is considered to be more stress-tolerant than cultivated soybean (G. max), and has considerable genetic variation for increasing alkalinity tolerance of soybean. In this study, we analyzed the transcriptome profile in the roots of an alkalinity tolerant wild soybean variety N24852 at 12 and 24 h after 90 mM NaHCO3 stress by RNA-sequencing. Compared with the controls, a total of 449 differentially expressed genes (DEGs) were identified, including 95 and 140 up-regulated genes, and 108 and 135 down-regulated genes at 12 and 24 h after NaHCO3 treatment, respectively. Quantitative RT-PCR analysis of 14 DEGs showed a high consistency with their expression profiles by RNA-sequencing. Gene Ontology (GO) terms related to transcription factors and transporters were significantly enriched in the up-regulated genes at 12 and 24 h after NaHCO3 stress, respectively. Nuclear factor Y subunit A transcription factors were enriched at 12 h after NaHCO3 stress, and high percentages of basic helix-loop-helix, ethylene-responsive factor, Trihelix, and zinc finger (C2H2, C3H) transcription factors were found at both 12 and 24 h after NaHCO3 stress. Genes related to ion transporters such as ABC transporter, aluminum activated malate transporter, glutamate receptor, nitrate transporter/proton dependent oligopeptide family, and S-type anion channel were enriched in up-regulated DEGs at 24 h after NaHCO3 treatment, implying their roles in maintaining ion homeostasis in soybean roots under alkalinity. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed “phenylpropanoid biosynthesis” and “phenylalanine metabolism” pathways might participate in soybean response to alkalinity. This study provides a foundation to further investigate the functions of NaHCO3 stress-responsive genes and the molecular basis of soybean tolerance to alkalinity. PMID:28018382
PsMPK7, a stress-associated mitogen-activated protein kinase (MAPK) in Phytophthora sojae, is required for stress tolerance, reactive oxygenated species detoxification, cyst germination, sexual reproduction and infection of soybean.
Gao, Jian; Cao, Mingna; Ye, Wenwu; Li, Haiyang; Kong, Liang; Zheng, Xiaobo; Wang, Yuanchao
The sensing of stress signals and their transduction into appropriate responses are crucial for the adaptation, survival and infection of phytopathogenic fungi and oomycetes. Amongst evolutionarily conserved pathways, mitogen-activated protein kinase (MAPK) cascades function as key signal transducers that use phosphorylation to convey information. In this study, we identified a gene, designated PsMPK7, one of 14 predicted genes encoding MAPKs in Phytophthora sojae. PsMPK7 was highly transcribed in each tested stage, but was up-regulated in the zoospore, cyst and cyst germination stages. Silencing of PsMPK7 affected the growth of germinated cysts, oospore production and the pathogenicity of soybean. PsMPK7 transcription was induced by stresses from sorbitol, NaCl and hydrogen peroxide. Transformants in which PsMPK7 expression was silenced (PsMPK7-silenced) were significantly more sensitive to osmotic and oxidative stress. Aniline blue and diaminobenzidine staining revealed that the silenced lines did not suppress the host reactive oxygen species (ROS) burst, indicating that either the inoculated plants activated stronger defence responses to the transformants and/or the PsMPK7-silenced transformants failed to overcome plant defences. In addition, extracellular secretion of laccase decreased in the silenced lines. Overall, our results indicate that the PsMPK7 gene encodes a stress-associated MAPK in P. sojae that is important not only for responses to various stresses, but also for ROS detoxification, cyst germination, sexual oospore production and infection of soybean.
Bruner, Robert F; Hodgson, Erin W; Gassmann, Aaron J
The Rag1 gene confers antibiotic resistance to soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), and in 2010, varieties expressing Rag1 were released for commercial use in the United States. We do not know how Rag1 varieties will influence the broader community of defoliating insects that inhabit soybean fields. In 2010 and 2011, the preference and performance of pest insects that defoliate soybeans [Glycines max (L.) Merr] were tested using Rag1 and aphid-susceptible varieties. Three coleopterans and four lepidopterans were used: northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae); southern corn rootworm, Diabrotica undecimpunctata howardi Barber (Coleoptera: Chrysomelidae); bean leaf beetle, Ceratoma trsifurcata Förster (Coleoptera: Chrysomelidae); fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae); corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae); soybean looper, Chrysodeix includens (Walker) (Lepidoptera: Noctuidae); and velvet-bean caterpillar, Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae). The preference of insects was evaluated in choice and no-choice tests using Rag1 and susceptible soybeans. Lepidopterans also were evaluated on Rag1 leaves using four nutritional indices: relative growth rate, approximate digestibility, and efficiency of conversion of ingested material. In the majority of preference tests, no effect of Rag1 was detected, and in cases where preferences were found, there was no consistent pattern of preference for Rag1 vs. susceptible leaf tissue. Helicoverpa zea demonstrated a preference for resistant leaf tissue, but this was dependent on the genetic background of the variety. Evaluations of nutritional indices indicated that three species of Lepidoptera, S. frugiperda, H. zea, and A. gemmatalis, displayed reduced conversion efficiency for Rag1 soybeans, suggesting effects of antibiosis.
Chen, Chao; Sun, Xiaoli; Duanmu, Huizi; Zhu, Dan; Yu, Yang; Cao, Lei; Liu, Ailin; Jia, Bowei; Xiao, Jialei; Zhu, Yanming
Calcium, as the most widely accepted messenger, plays an important role in plant stress responses through calcium-dependent signaling pathways. The calmodulin-like family genes (CMLs) encode Ca2+ sensors and function in signaling transduction in response to environmental stimuli. However, until now, the function of plant CML proteins, especially soybean CMLs, is largely unknown. Here, we isolated a Glycine soja CML protein GsCML27, with four conserved EF-hands domains, and identified it as a calcium-binding protein through far-UV CD spectroscopy. We further found that expression of GsCML27 was induced by bicarbonate, salt and osmotic stresses. Interestingly, ectopic expression of GsCML27 in Arabidopsis enhanced plant tolerance to bicarbonate stress, but decreased the salt and osmotic tolerance during the seed germination and early growth stages. Furthermore, we found that ectopic expression of GsCML27 decreases salt tolerance through modifying both the cellular ionic (Na+, K+) content and the osmotic stress regulation. GsCML27 ectopic expression also decreased the expression levels of osmotic stress-responsive genes. Moreover, we also showed that GsCML27 localized in the whole cell, including cytoplasm, plasma membrane and nucleus in Arabidopsis protoplasts and onion epidermal cells, and displayed high expression in roots and embryos. Together, these data present evidence that GsCML27 as a Ca2+-binding EF-hand protein plays a role in plant responses to bicarbonate, salt and osmotic stresses. PMID:26550992
Fiuza, Lidia Mariana
The bacterium Bacillus thuringiensis (Bt) produces delta-endotoxins that possess toxic properties and can be used as biopesticides, as well as a source of genes for the construction of transgenic plants resistant to insects. In Brazil, the introduction of Bt soybean with insecticidal properties to the velvetbean caterpillar, the main insect pest of soybean, has been seen a promising tool in the management of these agroecosystems. However, the increase in stink bug populations in this culture, in various regions of the country, which are not susceptible to the existing genetically modified plants, requires application of chemicals that damage the environment. Little is known about the actual toxicity of Bt to Hemiptera, since these insects present sucking mouthparts, which hamper toxicity assays with artificial diets containing toxins of this bacterium. In recent studies of cytotoxicity with the gut of different hemipterans, susceptibility in the mechanism of action of delta-endotoxins has been demonstrated, which can generate promising subsidies for the control of these insect pests in soybean. This paper aims to review the studies related to the selection, application and mode of action of Bt in the biological control of the major pest of soybean, Anticarsia gemmatalis, and an analysis of advances in research on the use of Bt for control hemipterans. PMID:24575310
Schünemann, Rogério; Knaak, Neiva; Fiuza, Lidia Mariana
The bacterium Bacillus thuringiensis (Bt) produces delta-endotoxins that possess toxic properties and can be used as biopesticides, as well as a source of genes for the construction of transgenic plants resistant to insects. In Brazil, the introduction of Bt soybean with insecticidal properties to the velvetbean caterpillar, the main insect pest of soybean, has been seen a promising tool in the management of these agroecosystems. However, the increase in stink bug populations in this culture, in various regions of the country, which are not susceptible to the existing genetically modified plants, requires application of chemicals that damage the environment. Little is known about the actual toxicity of Bt to Hemiptera, since these insects present sucking mouthparts, which hamper toxicity assays with artificial diets containing toxins of this bacterium. In recent studies of cytotoxicity with the gut of different hemipterans, susceptibility in the mechanism of action of delta-endotoxins has been demonstrated, which can generate promising subsidies for the control of these insect pests in soybean. This paper aims to review the studies related to the selection, application and mode of action of Bt in the biological control of the major pest of soybean, Anticarsia gemmatalis, and an analysis of advances in research on the use of Bt for control hemipterans.
Paulillo, Luis Cesar Maffei Sartini; Sebbenn, Alexandre Magno; de Carvalho Derbyshire, Maria Tereza Vitral; Góes-Neto, Aristóteles; de Paula Brotto, Marco Aurélio; Figueira, Antonio
We have characterized in vitro and in vivo effects of trypsin inhibitors from Theobroma seeds on the activity of trypsin- and chymotrypsin-like proteins from Lepidopteran pest insects. The action of semipurified trypsin inhibitors from Theobroma was evaluated by the inhibition of bovine trypsin and chymotrypsin activities determined by the hydrolysis of N-Benzoyl-DL-Arginine-p-Nitroanilide (BAPA) and N-Succinyl-Ala-Ala-Pho-Phe p-Nitroanilide (S-(Ala)2ProPhe-pNA). Proteinase inhibitor activities from Theobroma cacao and T. obovatum seeds were the most effective in inhibiting trypsin-like proteins, whereas those from T. obovatum and T. sylvestre were the most efficient against chymotrypsin-like proteins. All larvae midgut extracts showed trypsin-like proteolytic activities, and the putative trypsin inhibitors from Theobroma seeds significantly inhibited purified bovine trypsin. With respect to the influence of Theobroma trypsin inhibitors on intact insects, the inclusion of T. cacao extracts in artificial diets of velvet bean caterpillars (Anticarsia gemmatalis) and sugarcane borer (Diatraea saccharalis) produced a significant increase in the percentage of adult deformation, which is directly related to both the survival rate of the insects and oviposition.
Panizzi, A R
The integrated pest management (IPM) of soybean developed and implemented in Brazil was one of the most successful programs of pest management in the world. Established during the 1970s, it showed a tremendous level of adoption by growers, decreasing the amount of insecticide use by over 50%. It included outstanding approaches of field scouting and decision making, considering the economic injury levels (EILs) for the major pests. Two main biological control programs were highly important to support the soybean IPM program in Brazil, i.e., the use of a NPVAg to control the major defoliator, the velvet bean caterpillar, Anticarsia gemmatalis Hübner, and the use of egg parasitoids against the seed-sucking stink bugs, in particular, the southern green stink bug, Nezara viridula (L.). These two biological control programs plus pests scouting, and the use of more selective insecticides considering the EILs supported the IPM program through the 1980s and 1990s. With the change in the landscape, with the adoption of the no-tillage cultivation system and the introduction of more intense multiple cropping, and with the lower input to divulge and adapt the IPM program to this new reality, the program started to decline during the years 2000s. Nowadays, soybean IPM is almost a forgotten control technology. In this mini-review article, suggestions are made to possibly revive and adapt the soybean IPM to contemporary time.
Rodas, Valeria M; Marques, Fabiano H; Honda, Marcelo T; Soares, Daniela M; Jorge, Soraia A C; Antoniazzi, Marta M; Medugno, Claudia; Castro, Maria E B; Ribeiro, Bergmann M; Souza, Marlinda L; Tonso, Aldo; Pereira, Carlos A
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. We have assayed the k(L)a of the systems, initial cell seeding, cell culture volume, dissolved oxygen (DO), multiplicity of infection (MOI), nutrients consumption, and metabolites production. The medium surface oxygen transfer was shown to be higher in shaker bottles than in spinner ones, which was in direct correlation to the higher cell density obtained. Best quantitative performances of PIBs production were obtained with a SF900II medium volume/shaker-bottle volume ratio of 15% and MOI of 0.5 to 1 performed at a cell concentration at infection (CCI) of 1 to 2.5x10(6) cells/ml in a medium containing enough glucose and glutamine. Upon infection, a decrease in the cell multiplication was observed to be dependent on the MOI used, and the muX at the exponential growth phase in infected and non-infected cultures were, respectively, of 0.2832 and 0.3914 (day(-1)). The glucose consumption and lactate production were higher in the infected cultures (muGlucose and muLactate of, respectively, 0.0248 and 0.0089x10(-8) g/cellxday in infected cultures and 0.0151 and 0.0046x10(-8) g/cellxday in non infected ones). The glutamine consumption did not differ in both cultures (muGlutamine of 0.0034 and 0.0037x10(-8) g/cellxday in, respectively, infected and non infected cultures). When a virus MOI of 0.1 to 1 was used for infection, a higher concentration of PIBs/ml was obtained. This was in direct correlation to a higher cell concentration present in these cultures, where a decrease in cell multiplication due to virus infection is minimized. When a MOI of 1 was used, a more effective decrease in cell multiplication was observed and a lower concentration of PIBs/ml was obtained, but with the best
Eberhardt, Ignacio; Gioria, Verónica Viviana; Micheloud, Gabriela Analía; Claus, Juan Daniel
The infectivity of stocks of baculoviruses produced in serum-free media is sensitive to freezing at ultra-low temperatures. The objective of this work was to elucidate the causes of such sensitivity, using as a model the freezing of stocks of Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV), a baculovirus widely employed as biological insecticide. Titers of supernatants of cell cultures infected with AgMNPV in four different serum-free media supplemented with lipid emulsions were reduced by 50 to 90% after six months freezing. By using a full factorial experiment, freezing and lipid emulsion, as well as the interaction between them, were identified as the main factors reducing the viral titer. The virucidal effect of the lipid emulsion was reproduced by one of their components, the surfactant Polysorbate 80. Damaged viral envelopes were observed by transmission electron microscopy in most particles frozen in a medium supplemented with lipid emulsion or Polysorbate 80. Additionally, Polysorbate 80 also affected the infectivity of AgMNPV stocks that were incubated at 27°C. The identification of the roles played by the lipid emulsion and Polysorbate 80 is not only a contribution to the understanding of the mechanisms underlying the inactivation of baculovirus stocks produced in serum-free media during storage at ultra-low temperature, but is also an input for the rational development of new procedures aimed at improving both the preservation of baculovirus stocks and the composition of culture media for the production of baculovirus-based bioproducts in insect cells. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1559-1569, 2016.
Background The structure of regulatory networks remains an open question in our understanding of complex biological systems. Interactions during complete viral life cycles present unique opportunities to understand how host-parasite network take shape and behave. The Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) is a large double-stranded DNA virus, whose genome may encode for 152 open reading frames (ORFs). Here we present the analysis of the ordered cascade of the AgMNPV gene expression. Results We observed an earlier onset of the expression than previously reported for other baculoviruses, especially for genes involved in DNA replication. Most ORFs were expressed at higher levels in a more permissive host cell line. Genes with more than one copy in the genome had distinct expression profiles, which could indicate the acquisition of new functionalities. The transcription gene regulatory network (GRN) for 149 ORFs had a modular topology comprising five communities of highly interconnected nodes that separated key genes that are functionally related on different communities, possibly maximizing redundancy and GRN robustness by compartmentalization of important functions. Core conserved functions showed expression synchronicity, distinct GRN features and significantly less genetic diversity, consistent with evolutionary constraints imposed in key elements of biological systems. This reduced genetic diversity also had a positive correlation with the importance of the gene in our estimated GRN, supporting a relationship between phylogenetic data of baculovirus genes and network features inferred from expression data. We also observed that gene arrangement in overlapping transcripts was conserved among related baculoviruses, suggesting a principle of genome organization. Conclusions Albeit with a reduced number of nodes (149), the AgMNPV GRN had a topology and key characteristics similar to those observed in complex cellular organisms, which indicates
Lauzon, Hilary A M; Jamieson, Peter B; Krell, Peter J; Arif, Basil M
Two distinct nucleopolyhedrovirus species of the eastern spruce budworm, Choristoneura fumiferana, exist in a symbiont-like relationship. C. fumiferana defective nucleopolyhedrovirus (CfDEFNPV) only infects C. fumiferana larvae per os in the presence of C. fumiferana nucleopolyhedrovirus Ireland strain (CfMNPV), but is infective when injected into the haemolymph. CfDEFNPV synergizes CfMNPV in per os infections and CfMNPV is always the predominant progeny. This study was undertaken to report the genomic makeup and organization of CfDEFNPV in an attempt to identify its defect and understand its synergistic role. The genome was mapped, sequenced, characterized and compared to other baculoviruses. The CfDEFNPV genome was 131,160 nt long with 149 putative open reading frames (ORFs) and a G + C content of 45.8 mol%. Homologues of all 62 conserved lepidopteran baculovirus genes were found including those implicated in per os infectivity, p74, per os infectivity factor (pif) and pif-2. Although no obvious deletions were observed to explain the defect, two ORFs, Cfdef79 and Cfdef99 (inhibitor of apoptosis-4), contained potential deletions. Cfdef50 (late expression factor-10)/Cfdef51 (vp1054) and Cfdef76/Cfdef77 (telokin-like protein) had large overlaps and a potential homologue to ac105/he65 was split. Four baculovirus repeat ORFs were present, as were two unique genes, but no enhancins were identified. CfDEFNPV contained 13 homologous regions, each with one to five palindromes. Comparison with fully sequenced baculovirus genomes identified CfDEFNPV as a group I NPV with the closest average amino acid identity to Epiphyas postvittana NPV, followed by Orgyia pseudotsugata MNPV and CfMNPV, with its closest matches being to individual Anticarsia gemmatalis MNPV gene sequences.
de Oliveira, Edmar Justo; Rabinovitch, Leon; Monnerat, Rose Gomes; Passos, Liana Konovaloff Jannotti; Zahner, Viviane
Thirty-three strains of Brevibacillus laterosporus, including three novel strains isolated from Brazilian soil samples, were examined for genetic variability by the use of different PCR-based methods. Molecular markers that could characterize bacterial strains with regards to their pathogenic potential were investigated. In addition, toxicity was assessed by the use of insects belonging to the orders Lepidoptera and Coleoptera and the mollusk Biomphalaria glabrata. Among the targets tested, Biomphalaria glabrata demonstrated the highest degree of sensitivity to B. laterosporus, with some strains inducing 90 to 100% mortality in snails aged 3 and 12 days posteclosion. Larvae of the coleopteron Anthonomus grandis were also susceptible, presenting mortality levels of between 33 and 63%. Toxicity was also noted towards the lepidopteron Anticarsia gemmatalis. In contrast, no mortality was recorded among test populations of Tenebrio molitor or Spodoptera frugiperda. The application of intergenic transcribed spacer PCR and BOX-PCR generated 15 and 17 different genotypes, respectively. None of the molecular techniques allowed the identification of a convenient marker that was associated with any entomopathogenic phenotype. However, a 1,078-bp amplicon was detected for all strains of B. laterosporus when a primer for amplification of the BOXA1R region was used. Similarly, a 900-bp amplicon was generated from all isolates by use of the primer OPA-11 for randomly amplified polymorphic DNA analysis. These amplicons were not detected for other phenotypically related Brevibacillus species, indicating that they represent markers that are specific for B. laterosporus, which may prove useful for the isolation and identification of new strains of this species.
de Oliveira, Edmar Justo; Rabinovitch, Leon; Monnerat, Rose Gomes; Passos, Liana Konovaloff Jannotti; Zahner, Viviane
Thirty-three strains of Brevibacillus laterosporus, including three novel strains isolated from Brazilian soil samples, were examined for genetic variability by the use of different PCR-based methods. Molecular markers that could characterize bacterial strains with regards to their pathogenic potential were investigated. In addition, toxicity was assessed by the use of insects belonging to the orders Lepidoptera and Coleoptera and the mollusk Biomphalaria glabrata. Among the targets tested, Biomphalaria glabrata demonstrated the highest degree of sensitivity to B. laterosporus, with some strains inducing 90 to 100% mortality in snails aged 3 and 12 days posteclosion. Larvae of the coleopteron Anthonomus grandis were also susceptible, presenting mortality levels of between 33 and 63%. Toxicity was also noted towards the lepidopteron Anticarsia gemmatalis. In contrast, no mortality was recorded among test populations of Tenebrio molitor or Spodoptera frugiperda. The application of intergenic transcribed spacer PCR and BOX-PCR generated 15 and 17 different genotypes, respectively. None of the molecular techniques allowed the identification of a convenient marker that was associated with any entomopathogenic phenotype. However, a 1,078-bp amplicon was detected for all strains of B. laterosporus when a primer for amplification of the BOXA1R region was used. Similarly, a 900-bp amplicon was generated from all isolates by use of the primer OPA-11 for randomly amplified polymorphic DNA analysis. These amplicons were not detected for other phenotypically related Brevibacillus species, indicating that they represent markers that are specific for B. laterosporus, which may prove useful for the isolation and identification of new strains of this species. PMID:15528531
Schmelz, Eric A.; Huffaker, Alisa; Carroll, Mark J.; Alborn, Hans T.; Ali, Jared G.; Teal, Peter E.A.
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 γ-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-InΔV; +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 γ-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. PMID:23008466
Gao, Yue; Tao, Bo; Qiu, Lijuan; Jin, Longguo; Wu, Jing
The physiological mechanisms underlying glyphosate resistance in wild soybean germplasm and relevant EPSPS gene expression were evaluated. These germplasms were selected by gradually increasing glyphosate selection pressure started from 2010. As indicated by a whole-plant dose response bioassay, ZYD-254 plants were resistant to glyphosate at concentrations of 1230gaeha(-1), but the susceptible plants (ZYD-16) were unable to survive in the presence of 300gaeha(-1) glyphosate. The ED50 values of resistant germplasm were approximately 8.8 times of the susceptible germplasm. Chlorophyll content was significantly decreased in ZYD-16 plants in comparison with ZYD-254 plants. ZYD-16 plants accumulated 10.1 times more shikimate in leaves at 5days after glyphosate treatment at 1230gaeha(-1) than ZYD-254 did. GST activity differed between ZYD-254 and ZYD-16 in three tissues. It was highest in leaves. There were no significant differences in EPSPS1 or EPSPS3 expression between two germplasms before exposure to glyphosate treatment. After glyphosate treatment, there was a 2- to 4-fold increase in EPSPS1 mRNA levels in ZYD-254, but there was no change in EPSPS3 mRNA levels in ZYD-254 or ZYD-16.
Diverse pathogens secrete effector proteins into plant cells to manipulate host cellular processes. Oomycete pathogens contain very large complements of predicted effector genes defined by an RXLR host cell entry motif. The genome of Hyaloperonospora arabidopsidis (Hpa, downy mildew of Arabidopsis) ...
Transgenic soybean plants were generated using bombardment of embryogenic cultures with the PAL5 (phenylalanine ammonia lyase), CHS6 (chalcone synthase) and IFS2 (isoflavone synthase) genes in sense orientation, driven by the cotyledon-preferable lectin promoter, or with the IFS2 (isoflavone synthas...
Improving resistance for Phytophthora root and stem rot is an important goal in soybean [Glycine max (L.) Merr.] breeding. Partial resistance can be as effective in managing this disease as single-gene (Rps) mediated resistance and is more durable. The objective of this study was to identify QTL con...
The activity of anti-oxidant enzymes (Superoxide dismutase (SOD), Peroxidase (POD), Catalase (CAT) and parameters of oxidative stress malondialdehyde (MDA) of shoots were investigated in S. sieb naturally salt-resistant halophyte. The seedlings of S. sieb were treated with varying (0, 80, 160 and 240 mM) NaCl stress. The results showed that NaCl played an important role in growth of S. sieb. It made obviously promotion of certain NaCl concentration to growth of S. sieb, the seeflings of S. sieb grew best under 80 mM salt stress. MDA concentration of S. sieb obviously decreased under 80 mM salt stress then increased with salt concentration increased. The activities of SOD, POD and CAT increased with the increase of the concentration of NaCl in S. sieb. The salt tolerance of this halophyte under salt stress condition are probably due to its ability to exhibit high SOD, POD and CAT enzyme activities and Soluble Sugar (SS) concentration.
Wang, K J; Takahata, Y; Kono, Y; Kaizuma, N
Soybean Kunitz trypsin inhibitor (SKTI) has several polymorphic types, which are controlled by co-dominant multiple alleles at a single locus. Of these types, Tia and Tib are predominant types, and there are nine differences in amino acids between Tia and Tib. Recently, an intermediate transitional type (Tibi5) between them was detected. However, other transitional types have not been detected despite surveys of many cultivated and wild soybeans. One of the reasons why other transitional variants have not been found is inferred to be due to the difficulty of the detection of SKTI protein variants by polyacrylamide gel electrophoresis (PAGE). To detect novel variants of SKTI, nucleotide sequence analysis in addition to PAGE was carried out. Four new variants were found from many Japanese wild soybeans. Of these variants, three (designated as Tiaa1, Tiaa2, Tiab1) were detected through gene sequence analysis on wild soybeans having the same electrophoretic mobility as Tia, and one (Tig) was detected through PAGE. The Tig variant showed a slightly lower electrophoretic mobility than Tic. The nucleotide sequences of Tig were identical to those of Tib except for one T-->C transitional mutation at position +340. The sequences of Tiaa1 and Tiaa2 genes were identical to those of Tia with the exception of a G-->A mutation at position +376 and a T-->C mutation at +404, respectively. The sequence of Tiab1 differed from Tia by three nucleotides: C-->A at position +331, T-->C at +459 and A-->G at +484. Of the three nucleotide changes, two were common to Tiab1, Tibi5 and Tib, suggesting that Tiab1 is an intermediate transitional type between Tia and Tib. Our results suggest that Tib type has been differentiated through a series of mutations from Tia before the domestication of cultivated soybean.
Luo, Xiao; Sun, Xiaoli; Liu, Baohui; Zhu, Dan; Bai, Xi; Cai, Hua; Ji, Wei; Cao, Lei; Wu, Jing; Wang, Mingchao; Ding, Xiaodong; Zhu, Yanming
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.
He, Shui-Lian; Wang, Yun-Sheng; Li, De-Zhu; Yi, Ting-Shuang
Wild soybean, the direct progenitor of cultivated soybean, inhabits a wide distribution range across the mainland of East Asia and the Japanese archipelago. A multidisciplinary approach combining analyses of population genetics based on 20 nuclear microsatellites and one plastid locus were applied to reveal the genetic variation of wild soybean, and the contributions of geographical, environmental factors and historic climatic change on its patterns of genetic differentiation. High genetic diversity and significant genetic differentiation were revealed in wild soybean. Wild soybean was inferred to be limited to southern and central China during the Last Glacial Maximum (LGM) and experienced large-scale post-LGM range expansion into northern East Asia. A substantial northward range shift has been predicted to occur by the 2080s. A stronger effect of isolation by environment (IBE) versus isolation by geographical distance (IBD) was found for genetic differentiation in wild soybean, which suggested that environmental factors were responsible for the adaptive eco-geographical differentiation. This study indicated that IBE and historical climatic change together shaped patterns of genetic variation and differentiation of wild soybean. Different conservation measures should be implemented on different populations according to their adaptive potential to future changes in climate and human-induced environmental changes. PMID:26952904
Chen, Yun-Xia; Wu, Mian; Ma, Fang-Fang; Chen, Jian-Qun; Wang, Bin
Soybean mosaic virus (SMV) is a devastating plant virus classified in the family Potyviridae, and known to infect cultivated soybeans (Glycine max). In this study, seven new SMVs were isolated from wild soybean samples and analyzed by whole-genome sequencing. An updated SMV phylogeny was built with the seven new and 83 known SMV genomic sequences. Results showed that three northeastern SMV isolates were distributed in clade III and IV, while four southern SMVs were grouped together in clade II and all contained a recombinant BCMV fragment (~900 bp) in the upstream part of the genome. This work revealed that wild soybeans in China also act as important SMV hosts and play a role in the transmission and diversity of SMVs.
Phytopathogenic species of Diaporthe are associated with the serious diseases including seed decay, pod and stem blight and stem canker of soybean leading to considerable loss of crop production worldwide. Accurate identification of the species that cause these diseases has been difficult due to the...
He, Shui-Lian; Wang, Yun-Sheng; Li, De-Zhu; Yi, Ting-Shuang
Wild soybean, the direct progenitor of cultivated soybean, inhabits a wide distribution range across the mainland of East Asia and the Japanese archipelago. A multidisciplinary approach combining analyses of population genetics based on 20 nuclear microsatellites and one plastid locus were applied to reveal the genetic variation of wild soybean, and the contributions of geographical, environmental factors and historic climatic change on its patterns of genetic differentiation. High genetic diversity and significant genetic differentiation were revealed in wild soybean. Wild soybean was inferred to be limited to southern and central China during the Last Glacial Maximum (LGM) and experienced large-scale post-LGM range expansion into northern East Asia. A substantial northward range shift has been predicted to occur by the 2080s. A stronger effect of isolation by environment (IBE) versus isolation by geographical distance (IBD) was found for genetic differentiation in wild soybean, which suggested that environmental factors were responsible for the adaptive eco-geographical differentiation. This study indicated that IBE and historical climatic change together shaped patterns of genetic variation and differentiation of wild soybean. Different conservation measures should be implemented on different populations according to their adaptive potential to future changes in climate and human-induced environmental changes.
Krishnamurthy, Panneerselvam; Tsukamoto, Chigen; Takahashi, Yuya; Hongo, Yuji; Singh, Ram J; Lee, Jeong Dong; Chung, Gyuhwa
Eight wild soybean accessions with different saponin phenotypes were used to examine saponin composition and relative saponin quantity in various tissues of mature seeds and two-week-old seedlings by LC-PDA/MS/MS. Saponin composition and content were varied according to tissues and accessions. The average total saponin concentration in 1 g mature dry seeds of wild soybean was 16.08 ± 3.13 μmol. In two-week-old seedlings, produced from 1 g mature seeds, it was 27.94 ± 6.52 μmol. Group A saponins were highly concentrated in seed hypocotyl (4.04 ± 0.71 μmol). High concentration of DDMP saponins (7.37 ± 5.22 μmol) and Sg-6 saponins (2.19 ± 0.59 μmol) was found in cotyledonary leaf. In seedlings, the amounts of group A and Sg-6 saponins reduced 2.3- and 1.3-folds, respectively, while DDMP + B + E saponins increased 2.5-fold than those of mature seeds. Our findings show that the group A and Sg-6 saponins in mature seeds were degraded and/or translocated by germination whereas DDMP saponins were newly synthesized.
Zhang, Chunhong; Meng, Qingchang; Gai, Junyi; Yu, Deyue
The terminal step of soybean cysteine synthesis is catalyzed by O-acetylserine(thiol)lyase (OAS-TL, EC 126.96.36.199). In this study, we isolated and characterized an OAS-TL gene from a wild soybean material (designated as GsOAS-TL1). GsOAS-TL1 cDNA sequence showed strict conservation at both nucleotide and amino acid levels compared with that from cultivated soybean. Genomic structure analysis of GsOAS-TL1 indicated that it contained 10 exons and 9 introns in the coding region with conserved exon sizes and intron locations compared with Arabidopsis thaliana OAS-TL-like genes. Among the complete GsOAS-TL1 cDNA and three part-deletion fragments, only expression of the full-length cDNA could rescue the NK3 cys(-) Escherichia coli auxotroph, which was coherent with the assayed enzyme activity of purified fusion proteins. For RT-PCR analysis in different wild soybean tissues, GsOAS-TL1 showed lower expression in roots and developing seeds, whereas total OAS-TL activity of corresponding tissues showed significantly higher level in seeds than other tissues. To our knowledge, this is the first report on cloning and characterization of an OAS-TL gene from wild soybean. Our results are informative to further elucidate the function and evolution of OAS-TL in soybean.
not been well substantiated in the transfer of training literature (Burki-Cohen, Soja , & Longridge, 1998; Go, Burki-Cohen, & Soja , 2000; Go, Burki...504). Aberdeen Proving Ground: Army Research Laboratory. Burki-Cohen, J., Soja , N. N., & Longridge, T. (1998). Simulator platform motion - the need...driving simulator. Human Factors, 30(2), 201-217. Go, T. H., Burki-Cohen, J., & Soja , N. N. (2000). The effect of simulator motion on pilot training
The soybean aphid, Aphis glycines Matsumura, is the most important insect pest posing a threat to soybean, Glycine max (L.) Merr., grain production in the United States. Soybean cultivars with resistance are currently being deployed to aid in management of the pest. Temperature has been reported to ...
Soybean is a model for the legume research community due to its importance as a crop, a well populated genetic map, 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-b...
Tikhonov, A V; Nedoluzhko, A V; Martynov, V V; Dorokhov, D B
Data are presented on the genetic diversity and spatial structure of the natural wild soybean population from the neighborhood of the settlement of Ekaterinovka in Primorskii krai and on the relationship between the genetic structure of this population and its spatial organization. These data are discussed in comparison with the results of studies of wild soybean populations in the Far East region of the Russian Federation and China. Recommendations are given concerning the collection of genetic wild soybean resources.
sojae , a non- toxin-producing Aspergillus strain commonly used in the fer- mentation of soybeans to produce soy sauce and miso. The glyceollins...inoculated with Aspergillus sojae . After 3 days isoflavonoids were extracted from the inoculated seeds with 80% ethanol. The glyceollins were isolated...restriction mapping. Soybean treatment and harvesting A. sojae (SRRC1125) cultures were grown at 25 C in the dark on potato dextrose agar. After 5
effectiveness of motion in training simulators except for very specialized tasks and unique conditions (Bowen, Oakley & Barnett, 2006; Bürki-Cohen, Soja ...Arlington, VA: U.S. Army Research Institute for the Behavioral & Social Sciences.  Bürki-Cohen, J., Soja , N. N. & Longridge, R. (1998a). Simulator...Switzerland 216-231.  Bürki-Cohen, J., Soja , N. N. & Longridge, R. (1998b). Simulator Platform Motion—The Need Revisited. Paper presented at the
customer? The Federal Aviation Administration (FAA) has an alternative perspective on the criteria for choosing motion bases (Burki-Cohen, Soja ...series (Go, Burki-Cohen, & Soja , 2000; Longridge, Burki-Cohen, Go, & Kendra, 2001) addressed these issues: Are there any flight tasks for which a...Technical Report AL-TR- 1991-0139). Wright-Patterson AFB, OH: Air Force Materiel Command. 35 Burki-Cohen, J., Soja , N., & Longridge, T. (1998). Simulator
Xinjiang [Zhang Pingquan; NONGYE JINGJl WENTI, 23 Oct 87] 72 Analysis of Ecotypes of Wild Soybean (G. Soja ) in China [Xu Bao, ct at.; ZIIONGGUO NONGYE...should be suitably limited. 12986 Analysis of Ecotypes of Wild Soybean (G. Soja ) in China 40110011 Beijing ZHONGGUO NONGYE KEXUE [SCIENTIA...of Agricultural Sciences, Gongzhuling, Jilin] [Text] In 1981-1985, the photo-thermo effects on the development of wild soybean (G. soja Sieb, et
Phytophthora root and stem rot of soybean, caused by the oomycete, Phytophthora sojae, is one of the most destructive diseases to limit soybean production in the US. Although fourteen resistance genes (Rps) to P. sojae have been identified, adaptation of by the pathogen has made many of these ineffe...
Phytophthora root rot (PRR), caused by the oomycete Phytophthora sojae, is a devastating disease in soybean production. Using resistant cultivars has been suggested as the best solution for disease management. Michigan elite soybean E00003 is resistant to P. sojae and has been used as a PRR resist...
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 dynamic…
Helsinki, Finland, PAL 2210/92, November 16, 1992. 57 23. Soja , E., "DGME Waterfog Trials," British Ministry of Defense. Navy, YARD Report No. 4175...Firefighting Arrangements for Machinery Spaces - Comparisons and Evaluation," AMEE 2/75/3842, January 1975. Soja , E. (1990), "DGME Waterfog Trials," British
15, 2008. http://allafrica.com/stories/200809160135.html (accessed January 23, 2010), 119 Morray Boima. " Sojas Riot in Sierra Leone." Awareness...Occasional Paper Series: Volume 3, Number 2, 2008, 10. Boima, Morray. " Sojas Riot in Sierra Leone." Awareness Times, July 30, 2008. http://news.sl
electrophoresis. Proteomics 2:1169-1186. Chang, P. 2004. Lack of interaction between AFLR and AFLJ contributes to nonaflatoxigenicity of Aspergillus sojae ...nonaflatoxigenicity of Aspergillus sojae . Journal of Biotechnology 107: 245-253. Doyle, J. J., and J. L. Doyle. 1990. Isolation of plant DNA from fresh tissue. Focus
Kuroda, Yosuke; Kaga, Akito; Tomooka, Norihiko; Yano, Hiroshi; Takada, Yoshitake; Kato, Shin; Vaughan, Duncan
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.
der Waffen-S[., p. 95. 21 42. Arnold Purre, "Saksa-NÖuk. Liidu soja uldjooni," in Eestl Rllk ja Rahvas Telses Maallmasojas, Vol. 7, ed. by...Purre, "Saksa-Nouk. Liidu söja uldjooni," p. 35. 45. Hausser, pp. 95-96. 46. Purre, "Saksa-Nouk. Liidu soja uldjooni," p. 35. 47. Ibid. p. 36...of Estot.ia and life under the Soviet occupation.) 10. Maasing, Richard, et al. Ecsti Rilk Ja Rahvas Telses Maallma- sojas . Vol. 1
Liquid Chromatography Electrospray Ionization Mass Spectrometric (LC-ESI-MS) and Desorption Electrospray Ionization Mass Spectrometric (DESI-MS) Identification of Chemical Warfare Agents in Consumer Products
au-dessus des 6chantillons de semoule de mais et d’huile de soja ensemenc6s ont &6 6chantillonn6s avec des fibres SPME; ces fibres ont &6 analys6s au...6chantillons d’eau embouteill6e. Les vides au-dessus des 6chantillons de semoule de mais et d’huile de soja ensemencds ont 6t6 6chantillonn~s avec...des fibres SPNM. L’analyse directe des fibres SPME ayant 6t6 exposes dans le vide au-dessus des 6chantillons de la semoule de mais et I’huile de soja
Multiple disease resistance is an important component of production agriculture. Major challenges include resistance to Phytophthora root rot caused by evolving Phytophthora sojae races and the recently introduced invasive Asian soybean rust (ASBR) caused by Phakopsora pachyrhizi. The diseases cause...
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, while Glycine soja, the wild progenitor of soybean, is indeterminate. Indeterminate (Dt1) and determina...
a 33 1A. V .. In AC B . both A and ( SOJA can be used in the derivation of a. Note that this rule is applicable to the fictional agent So. Because... SoJA , A, r ’i a 3. A set of sound sequent rules r(v) for each view v which contains the axiom Id, and for which the rule Cut* is admissible. Also, r(v
n-l,n-p)) (4-7)n=0 Maximization of p( Soja ,g,si) with respect to a and sI is equivalent to minimizing z given by 1 N-1 TEp -7 . (s(n) - a .s(n-l,n-o...g and s are known. From Bayes’ rule, p(s Ia) p(a) p(as = P(o ) (4-21) and since g and sI are assumed known p(S oa) represents p( soja ,g,si) evaluated...at g and s, equal to their respective assumed known values. Therefore maximizing p(ai1O) is equivalent to maximizing p( SoJa )-p( a). Assuming p(a) is
Yuan, C P; Wang, Y J; Zhao, H K; Zhang, L; Wang, Y M; Liu, X D; Zhong, X F; Dong, Y S
Over-utilization of germplasms that are resistant to the soybean cyst nematode (SCN) in soybean breeding programs can lead to genetic vulnerability in resistant cultivars. Resistant wild soybean (Glycine soja) is considered an invaluable gene source for increasing the genetic diversity of SCN resistance. In this study, we genotyped 23 G. soja accessions that are resistant to SCN race 3 for polymorphisms in the resistance genes, rhg1, Rhg4, and SHMT, and investigated their genetic relationship with eight Glycine max resistant cultivars. We identified 89 single nucleotide polymorphisms (SNPs) and 11 DNA insertion-deletions (InDels), of which 70 SNPs and 8 InDels were found in rhg1, 9 SNPs were found in Rhg4, and 10 SNPs and 3 InDels were found in SHMT. Nucleotide diversity was π = 0.00238 and θ = 0.00235, and haplotype diversity was 1.000. A phylogenetic tree comprising four clusters was constructed using sequence variations of the 23 G. soja and 8 G. max resistant accessions. Five G. soja accessions in subcluster A2, and four G. soja accessions in cluster B were genetically distant from G. max genotypes. Eight resistance-associated SNPs in the three resistance genes formed nine haplotypes in total. G. soja resistant accessions had different haplotypes (H2, H4, H5, H6, H7, and H8) compared with those of G. max (H1, H3, and H9). These results provide vital information on the use of wild soybeans for broadening the genetic base of SCN resistance.
This paper draws on a study of a community-based adult education initiative, "Cumbria Credits," which took place during the period of serious economic decline which hit sections of the farming and the wider community in Cumbria during 2001. It draws on the principles underpinning Edward Soja's notion of "spatial justice" to explore transformations…
Yuzuki, Masanobu; Matsushima, Kenichiro; Koyama, Yasuji
Several key hydrolases in soy sauce fermentation such as proteases, peptidases, and glutaminases are supplied by Aspergillus sojae or Aspergillus oryzae. The genes encoding these hydrolases were successfully expressed in salt-tolerant yeast Zygosaccharomyces rouxii. These transformants are expected to supply extra hydrolases during soy sauce fermentation process.
Rubel, Laurie; Lim, Vivian; Hall-Wieckert, Maren; Katz, Sara
This paper explores teaching mathematics for spatial justice (Soja, 2010), as an extension of teaching mathematics for social justice (Gutstein, 2006). The study is contextualized in a 10-session curricular module focused on the spatial justice of a city's two-tiered system of personal finance institutions (mainstream vs. alternative), piloted…
Phytophthora root rot (PRR) caused by Phytophthora sojae is one of the most important soil-borne diseases in many soybean-production regions in the world. Identification of resistant gene(s) is an effective way for breeding to prevent soybean from being harmed by this disease. Here, two soybean popu...
The complete genomes of Phytophthora ramorum and P. sojae were sequenced in 2004. Two obvious questions arise, What contributions does the availability of a genome sequence make toward understanding the biology of Phytophthora spp.? What are the implications for management of sudden oak death in the...
Phytophthora root and stem rot (PRR) disease, caused by P. sojae, is a widespread soybean disease resulting in an annual yield loss of $1~2 billion worldwide. To control the disease, breeders primarily employ race-specific resistant genes which are named Rps genes which have been identified to be lo...
Whole and partial genome sequences are becoming available at an ever-increasing pace. For many plant pathogen systems, we are moving into the era of genome resequencing. The first Phytophthora genomes, P. ramorum and P. sojae, became available in 2004, followed shortly by P. infestans in 2006. Ava...
Phakopsora pachyrhizi, the causal organism of soybean rust, was first described in 1903 from leaves of Glycine max subsp. soja, or wild soybean, in Japan. Since that time, there have been numerous reports of the pathogen on various leguminous species around the world, first in Asia, followed by Aust...
Identification of resistance to reniform nematode (Rotylenchulus reniformis) is the first step in developing resistant soybean (Glycine max) cultivars that will benefit growers in the Mid South. This study was conducted to identify soybean (G. max and G. soja) lines with resistance to this pathogen....
Brooke, Robert; Coyle, Deborah; Walden, Anne; Healey, Conniem; Larson, Kim; Laughridge, Virginia; Ridder, Kim; Williams, Molly; Williams, Shawn
This article describes a teacher study group focusing on After School Writing Circles for elementary students as a site of Thirdspace professional development. Borrowing the concept of Thirdspace from postmodern geographer Edward Soja, the authors argue that professional development works best when teachers engage in the dual work of imagining and…
Vadeboncoeur, Jennifer A.
Drawing upon the concept of "thirdspace" (Soja 1996), this article extends sociocultural theorizations of space in relation to alternative educational programs: programs designed to re-engage youth who have been pushed out of mainstream schools. Snapshots of educational programs, provided by ethnographic research gathered in the United…
Similar to Arabidopsis, the wild soybean (Glycine soja) and many soybean (Glycine max) cultivars exhibit indeterminate stem growth controlled by a gene Dt1 – the functional counterpart of the Arabidopsis TFL1. Mutations in TFL1 and Dt1 both result in the shoot apical meristem (SAM) switching from ve...
Foxglove aphid, Aulacorthum solani (Kaltenbach), is a Hemipteran insect that infected a wide variety of plants worldwide and caused serious yield losses in crops. The objective of this study was to identify the putative QTL for foxglove aphid resistance in wild soybean, PI 366121, (Glycine soja Sieb...
Seventy-four wild and domestic soybean (Glycine max and G. soja) lines were evaluated for resistance to reniform nematode (Rotylenchulus reniformis) in growth chamber tests with a day length of 16 hours and temperature held constant at 28 C. Several entries for which reactions to reniform nematode w...
decrease in-orbital inclination a1A..40- .. Iv/"IY- t" east sal Wind 2020./a, at & MaM height of Sb~i 14# ., was 01-M2 he soja activity vat high-, and thelttd o eMl from I to 64S~ N -W I
Wild soybean (G. soja Sieb. & Zucc.) is the progenitor of soybean and is native to China, Taiwan, Japan, eastern Russia and the Korean peninsula. Research has repeatedly demonstrated that wild soybean is more genetically diverse than the cultivated soybean. There are 26 perennial Glycine species tha...
Imai, Mutsumi; Gentner, Dedre
Investigated whether learning the distinction between substance names and object names is conceptually or linguistically driven, by repeating Soja et al.'s study with English- and Japanese-speaking children. (Japanese does not make the count-mass grammatical distinction proposed to contribute to learning the distinction.) Found evidence for…
This article is essentially written as two linked parts. The first part considers how space, spatiality and history can contribute to understanding and "doing something about" the sustainability of rural communities. This is done by extensive reference to Soja's (1989 & 1996) space and spatial theorising and selective perspectives of…
DNA fingerprinting was performed on 64 strains of Aspergillus oryzae and one strain of A. sojae isolated from soysauce factories within Malaysia and Southeast Asia that use primitive traditional methods in producing 'tamari type' Cantonese soy sauce. PstI digests of total genomic DNA from each isol...
Space is related to power in that it is critical to the social production and reproduction of difference. This paper re-imagines a critical multiculturalism that embraces critical spatial theory and postmodern identity politics. In an overview of postmodern spatial theory, the works of Lefebvre (1974), E. Soja (1989, 1996), and M. Foucault (1986)…
Fedukovich, Casie J.
This article presents the National Day on Writing (NDOW) as an annual opportunity for writing programs to create program-supported thirdspaces (Burns; Mauk; Soja) that allow first-year writing students a laboratory in which to practice their developing rhetorical skills. The author proposes a contracted view of Ann Feldman's definition of…
Johnston, Jennifer A.
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…
DuanMu, Huizi; Wang, Yang; Bai, Xi; Cheng, Shufei; Deyholos, Michael K; Wong, Gane Ka-Shu; Li, Dan; Zhu, Dan; Li, Ran; Yu, Yang; Cao, Lei; Chen, Chao; Zhu, Yanming
Soil alkalinity is an important environmental problem limiting agricultural productivity. Wild soybean (Glycine soja) shows strong alkaline stress tolerance, so it is an ideal plant candidate for studying the molecular mechanisms of alkaline tolerance and identifying alkaline stress-responsive genes. However, limited information is available about G. soja responses to alkaline stress on a genomic scale. Therefore, in the present study, we used RNA sequencing to compare transcript profiles of G. soja root responses to sodium bicarbonate (NaHCO3) at six time points, and a total of 68,138,478 pairs of clean reads were obtained using the Illumina GAIIX. Expression patterns of 46,404 G. soja genes were profiled in all six samples based on RNA-seq data using Cufflinks software. Then, t12 transcription factors from MYB, WRKY, NAC, bZIP, C2H2, HB, and TIFY families and 12 oxidation reduction related genes were chosen and verified to be induced in response to alkaline stress by using quantitative real-time polymerase chain reaction (qRT-PCR). The GO functional annotation analysis showed that besides "transcriptional regulation" and "oxidation reduction," these genes were involved in a variety of processes, such as "binding" and "response to stress." This is the first comprehensive transcriptome profiling analysis of wild soybean root under alkaline stress by RNA sequencing. Our results highlight changes in the gene expression patterns and identify a set of genes induced by NaHCO3 stress. These findings provide a base for the global analyses of G. soja alkaline stress tolerance mechanisms.
Del Pozo-Valdivia, Alejandro I; Reisig, Dominic D; Bacheler, Jack S
Megacopta cribraria (F.), also known as the kudzu bug, is a soybean pest in the United States, and it can cause up to a 60% yield reduction if not controlled. Insecticides are commonly used to manage this pest in commercial soybean fields. However, other soybean production practices may also affect kudzu bug populations. This study investigated the effect of soil tillage, maturity group selection, and insecticide use on kudzu bug densities in soybean. During 2012 and 2013, at two locations each year in North Carolina, four varieties of soybean maturity groups were planted in June into conventionally tilled plots and into plots with cereal crop residue under reduced tillage conditions (mimicking double-crop production). Plots were further split as insecticide-protected and untreated. Four times more kudzu bugs were found in conventionally tilled than reduced till plots throughout the growing season. Selection of the maturity group influenced the attractiveness of the kudzu bug to oviposit on soybean. A 56% reduction of kudzu bug densities was achieved through insecticide treatment, with an ∼6% increase in yield. Information on how production practices, including soil tillage, affect kudzu bug populations in soybean may help growers select practices to minimize kudzu bug injury and protect yield.ResumenMegacopta cribraria (F.), conocido como el bicho del kudzu, es una plaga de la soja en los Estados Unidos que puede reducir el rendimiento hasta un 60% si no es controlado oportunamente con insecticidas en campos comerciales de soja. Sin embargo, otras prácticas de producción en la soja pueden afectar las poblaciones del bicho del kudzu. El presente estudio investigó el efecto de la labranza del campo, la selección del grupo de madurez en soja, y el uso de insecticidas en la densidad del bicho del kudzu. Durante el 2012 y el 2013, cuatro diferentes grupos de madurez de soja fueron plantados en Junio bajo el sistema tradicional de labranza y con labranza reducida
Costanzo, Stefano; Ospina-Giraldo, M D; Deahl, K L; Baker, C J; Jones, Richard W
A total of 18 paralogs of xyloglucan-specific endoglucanases (EGLs) from the glycosyl hydrolase family 12 were identified and characterized in Phytophthora sojae and Phytophthora ramorum. These genes encode predicted extracellular enzymes, with sizes ranging from 189 to 435 amino acid residues, that would be capable of hydrolyzing the xyloglucan component of the host cell wall. In two cases, four and six functional copies of these genes were found in tight succession within a region of 5 and 18 kb, respectively. The overall gene copy number and relative organization appeared well conserved between P. sojae and P. ramorum, with apparent synteny in this region of their respective genomes. Phylogenetic analyses of Phytophthora endoglucanases of family 12 and other known members of EGL 12, revealed a close relatedness with a fairly conserved gene sub-family containing, among others, sequences from the fungi Emericella desertorum and Aspergillus aculeatus. This is the first report of family 12 EGLs present in plant pathogenic eukaryotes.
Lamour, Kurt H; McDonald, W Hayes; Savidor, Alon
Genome sequences of the soybean pathogen, Phytophthora sojae, and the sudden oak death pathogen, Phytophthora ramorum, suggest a photosynthetic past and reveal recent massive expansion and diversification of potential pathogenicity gene families. Abstract: Draft genome sequences of the soybean pathogen, Phytophthora sojae, and the sudden oak death pathogen, Phytophthora ramorum, have been determined. O mycetes such as these Phytophthora species share the kingdom Stramenopila with photosynthetic algae such as diatoms and the presence of many Phytophthora genes of probable phototroph origin support a photosynthetic ancestry for the stramenopiles. Comparison of the two species' genomes reveals a rapid expansion and diversification of many protein families associated with plant infection such as hydrolases, ABC transporters, protein toxins, proteinase inhibitors and, in particular, a superfamily of 700 proteins with similarity to known o mycete avirulence genes.
Seversike, Thomas M; Sermons, Shannon M; Sinclair, Thomas R; Carter, Thomas E; Rufty, Thomas W
A key strategy in soybean drought research is increased stomatal sensitivity to high vapor pressure deficit (VPD), which contributes to the 'slow wilting' trait observed in the field. These experiments examined whether temperature of the growth environment affected the ability of plants to respond to VPD, and thus control transpiration rate (TR). Two soybean [Glycine max (L.) Merr.] and four wild soybean [Glycine soja (Sieb. and Zucc.)] genotypes were studied. The TR was measured over a range of VPD when plants were growing at 25 or 30°C, and again after an abrupt increase of 5°C. In G. max, a restriction of TR became evident as VPD increased above 2.0 kPa when temperature was near its growth optimum of 30°C. 'Slow wilting' genotype plant introduction (PI) 416937 exhibited greater TR control at high VPD compared with Hutcheson, and only PI 416937 restrained TR after the shift to 35°C. Three of the four G. soja genotypes exhibited control over TR with increasing VPD when grown at 25°C, which is near their estimated growth optimum. The TR control became engaged at lower VPD than in G. max and was retained to differing degrees after a shift to 30°C. The TR control systems in G. max and G. soja clearly were temperature-sensitive and kinetically definable, and more restrictive in the 'slow wilting' soybean genotype. For the favorable TR control traits observed in G. soja to be useful for soybean breeding in warmer climates, the regulatory linkage with lower temperatures must be uncoupled.
Soares da Costa, Carlos Alberto; De Sousa dos Santos, Aline; Santana Carlos, Aluana; Lopes Gonzalez, Gabrielle de Paula; Pontes Gaspar Reis, Rejane; Carneiro, Cynthia; Soares Alves, Sanderson; Pereira Albuquerque, Karine; Alves da Silva, Paula Cristina; Cavalcante Ribeiro, Danielle; Teles Boaventura, Gilson; de Moura, Egberto Gaspar; Alves do Nascimento-Saba, Celly Cristina
Introducción: El papel del ácido graso en la prevención o la progresión de las enfermedades crónicas ha generado un interés significativo por parte de los investigadores. Por lo tanto, nuestro objetivo fue evaluar los efectos a largo plazo de la dieta alta en grasas que contienen soja o aceite de canola en los parámetros de desarrollo del cuerpo y los huesos de ratas macho. Métodos: Después del destete, las ratas se agruparon y se alimentaron con una dieta control (7S) o una dieta alta en grasa que contiene soja (19S) o aceite de canola (19C). Fémur y vértebras lumbares (LV4) estructura se determinaron a los 180 días por absorciometría dual de rayos X y tomografía computarizada. Resultados: Los grupos mostraron similares ingesta de alimentos, la masa corporal y el desarrollo de longitud. Los parámetros óseos de la 19C fueron similares al grupo control, mientras que los 19S mostró parámetros óseos inferiores en comparación con los otros grupos. Conclusiones: La dieta alta en grasas que contiene aceite de soja fue desfavorable a la estructura ósea, mientras que el aceite de canola contribuyó salud de los huesos en la etapa adulta de la vida.
Ryba-White, M.; Nedukha, O.; Hilaire, E.; Guikema, J. A.; Kordyum, E.; Leach, J. E.; Spooner, B. S. (Principal Investigator)
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.
Franco-Rivera, Alejandro; Benintende, Graciela; Cozzi, Jorge; Baizabal-Aguirre, Victor Manuel; Valdez-Alarcón, Juan José; López-Meza, Joel Edmundo
Bacillus thuringiensis INTA 7-3, INTA 51-3, INTA Mo9-5 and INTA Mo14-4 strains were obtained from Argentina and characterized by determination of serotype, toxicity, plasmid composition, insecticidal gene content ( cry and vip ) and the cloning of the single- vip3A gene of the INTA Mo9-5 strain. The serotype analysis identified the serovars tohokuensis and darmstadiensis for the INTA 51-3 and INTA Mo14-4 strains, respectively, whereas the INTA Mo9-5 strain was classified as "autoagglutinated". In contrast to the plasmid patterns of INTA 7-3, INTA 51-3 and INTA Mo9-5 (which were similar to B. thuringiensis HD-1 strain), strain INTA Mo14-4 showed a unique plasmid array. PCR analysis of the four strains revealed the presence of cry genes and vip3A genes. Interestingly, it was found that B. thuringiensis 4Q7 strain, which is a plasmid cured strain, contained vip3A genes indicating the presence of these insecticidal genes in the chromosome. Bioassays towards various lepidopteran species revealed that B. thuringiensis INTA Mo9-5 and INTA 7-3 strains were highly active. In particular, the mean LC(50) obtained against A. gemmatalis larvae with the INTA Mo9-5 and INTA 7-3 strains were 7 (5.7-8.6) and 6.7 (5.6-8.0) ppm, respectively. The INTA Mo14-4 strain was non-toxic and strain INTA 51-3 showed only a weak larvicidal activity.
Nedukha, O.; Kordyum, E.; Leach, J.; Martyn, G.; Ryba-White, M.
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.
Bremer, H J; Anninos, A; Schulz, B
The amino acid composition of food products frequently used in the diets of amino acid and protein disorders-including tryptophan- was estimated using ion-exchange column chromatography and high performance liquid chromatography. It includes fruits (different varieties of apples, pears, ananas, bananas, peach, strawberries, honey melon, water melon, kiwi, plums, grapes), vegetables (different varieties of potatoes, potato products, cauli-flower, broccoli, cabbages, spinach, olives, lettuce, cucumber, peas, mushrooms) and commercially available or home-made food products (meat broth, fine gravy paste, ketchup, liquid seasoning, soja sauce, different varieties of Chinese noodles, sausages for phenylketonuria patients), and different new fiber concentrates.
Wang, S. E. Lofland, Z. Ma, L. Mohaddes-Ardabili, T . Zhao, L. Salamanca-Riba, S. R. Shinde , S. B. Ogale, F. Bai, D. Viehland, Y. Jia, D. G. Schlom, M...2008). 6S. S. Nonnenmann, E. M. Gallo, M. T . Coster, G. R. Soja, C. L. Johnson, R. S. Joseph, and J . E. Spanier, Appl. Phys. Lett. 95, 232903 (2009...7S. S. Nonnenmann, O. D. Leaffer, E. M. Gallo, M. T . Coster, and J . E. Spanier, Nano Lett. 10, 542 (2010). 8N. B. Chaure, P. Stamenov, F. M. F. Rhen
Tyler, Brett M.; Tripathy, Sucheta; Zhang, Xuemin; Dehal, Paramvir; Jiang, Rays H. Y.; Aerts, Andrea; Arredondo, Felipe D.; Baxter, Laura; Bensasson, Douda; Beynon, JIm L.; Chapman, Jarrod; Damasceno, Cynthia M. B.; Dorrance, Anne E.; Dou, Daolong; Dickerman, Allan W.; Dubchak, Inna L.; Garbelotto, Matteo; Gijzen, Mark; Gordon, Stuart G.; Govers, Francine; Grunwald, NIklaus J.; Huang, Wayne; Ivors, Kelly L.; Jones, Richard W.; Kamoun, Sophien; Krampis, Konstantinos; Lamour, Kurt H.; Lee, Mi-Kyung; McDonald, W. Hayes; Medina, Monica; Meijer, Harold J. G.; Nordberg, Erik K.; Maclean, Donald J.; Ospina-Giraldo, Manuel D.; Morris, Paul F.; Phuntumart, Vipaporn; Putnam, Nicholas J.; Rash, Sam; Rose, Jocelyn K. C.; Sakihama, Yasuko; Salamov, Asaf A.; Savidor, Alon; Scheuring, Chantel F.; Smith, Brian M.; Sobral, Bruno W. S.; Terry, Astrid; Torto-Alalibo, Trudy A.; Win, Joe; Xu, Zhanyou; Zhang, Hongbin; Grigoriev, Igor V.; Rokhsar, Daniel S.; Boore, Jeffrey L.
Draft genome sequences have been determined for the soybean pathogen Phytophthora sojae and the sudden oak death pathogen Phytophthora ramorum. Oömycetes such as these Phytophthora species share the kingdom Stramenopila with photosynthetic algae such as diatoms, and the presence of many Phytophthora genes of probable phototroph origin supports a photosynthetic ancestry for the stramenopiles. Comparison of the two species' genomes reveals a rapid expansion and diversification of many protein families associated with plant infection such as hydrolases, ABC transporters, protein toxins, proteinase inhibitors, and, in particular, a superfamily of 700 proteins with similarity to known oömycete avirulence genes.
Song, Tianqiao; Ma, Zhenchuan; Shen, Danyu; Li, Qi; Li, Wanlin; Su, Liming; Ye, Tingyue; Zhang, Meixiang; Wang, Yuanchao; Dou, Daolong
Oomycete pathogens produce a large number of CRN effectors to manipulate plant immune responses and promote infection. However, their functional mechanisms are largely unknown. Here, we identified a Phytophthora sojae CRN effector PsCRN108 which contains a putative DNA-binding helix-hairpin-helix (HhH) motif and acts in the plant cell nucleus. Silencing of the PsCRN108 gene reduced P. sojae virulence to soybean, while expression of the gene in Nicotiana benthamiana and Arabidopsis thaliana enhanced plant susceptibility to P. capsici. Moreover, PsCRN108 could inhibit expression of HSP genes in A. thaliana, N. benthamiana and soybean. Both the HhH motif and nuclear localization signal of this effector were required for its contribution to virulence and its suppression of HSP gene expression. Furthermore, we found that PsCRN108 targeted HSP promoters in an HSE- and HhH motif-dependent manner. PsCRN108 could inhibit the association of the HSE with the plant heat shock transcription factor AtHsfA1a, which initializes HSP gene expression in response to stress. Therefore, our data support a role for PsCRN108 as a nucleomodulin in down-regulating the expression of plant defense-related genes by directly targeting specific plant promoters.
Chaabane, M; Bidat, E; Chevallier, B
We report a case of food protein-induced enterocolitis syndrome (FPIES) with milk whose signs of milk intolerance began in the 1st days of life, consisting in minor and nonspecific symptoms. The 3 foods in question were cow's milk, soja, and wheat. The diagnosis of FPIES was suspected at the age of 9 months, after 3 hospitalizations for vomiting, sometimes associated with lethargy and hypotension, which occurred around 2h after cow's milk ingestion. Symptoms were not associated with positive specific IgE and cutaneous tests. Signs then occurred with soja and wheat. Because of the late diagnosis, 3 anaphylactic shock episodes occurred. FPIES is an uncommon cell-mediated food allergy reaction. This syndrome is characterized by gastrointestinal symptoms, especially severe vomiting, sometimes associated with anaphylactic shock. Usually signs occur 2h after ingestion. These reactions begin early, in the 1st months of life, and regress by the age of 3 years in 38-100% of cases depending on the responsible food. They are usually induced by cow's milk and soy proteins. Diagnosis is difficult and delayed because of nonspecific symptoms. Oral food challenge is the only examination that confirms the diagnosis. Treatment involves the exclusion of the specific food involved. Severe reactions require treatment of shock and adjunction of corticosteroids.
Vetukuri, Ramesh R; Asman, Anna Km; Jahan, Sultana N; Avrova, Anna O; Whisson, Stephen C; Dixelius, Christina
Advances in genome sequencing technologies have enabled generation of unprecedented information on genome content and organization. Eukaryote genomes in particular may contain large populations of transposable elements (TEs) and other repeated sequences. Active TEs can result in insertional mutations, altered transcription levels and ectopic recombination of DNA. The genome of the oomycete plant pathogen, Phytophthora infestans, contains vast numbers of TE sequences. There are also hundreds of predicted disease-promoting effector proteins, predominantly located in TE-rich genomic regions. Expansion of effector gene families is also a genomic signature of related oomycetes such as P. sojae. Deep sequencing of small RNAs (sRNAs) from P. infestans has identified sRNAs derived from all families of transposons, highlighting the importance of RNA silencing for maintaining these genomic invaders in an inactive form. Small RNAs were also identified from specific effector encoding genes, possibly leading to RNA silencing of these genes and variation in pathogenicity and virulence toward plant resistance genes. Similar findings have also recently been made for the distantly related species, P. sojae. Small RNA "hotspots" originating from arrays of amplified gene sequences, or from genes displaying overlapping antisense transcription, were also identified in P. infestans. These findings suggest a major role for RNA silencing processes in the adaptability and diversification of these economically important plant pathogens. Here we review the latest progress and understanding of gene silencing in oomycetes with emphasis on transposable elements and sRNA-associated events.
Ospina-Giraldo, Manuel D; McWalters, Jessica; Seyer, Lauren
The plant cell cuticle is the first obstacle for penetration of the host by plant pathogens. To breach this barrier, most pathogenic fungi employ a complex assortment of cell wall-degrading enzymes including carbohydrate esterases, glycoside hydrolases, and polysaccharide lyases. We characterized the full complement of carbohydrate esterase-coding genes in three Phytophthora species and analyzed the expression of cutinase in vitro and in planta; we also determined the cutinase allele distribution in multiple isolates of P. infestans. Our investigations revealed that there are 49, 21, and 37 esterase homologs in the P. infestans, P. ramorum, and P. sojae genomes, respectively, with a considerable number predicted to be extracellular. Four cutinase gene copies were found in both the P. infestans and P. ramorum genomes, while 16 copies were found in P. sojae. Transcriptional analyses of cutinase in P. infestans revealed that its expression level during infection is significantly upregulated at all time points compared to that of the same gene in mycelium grown in vitro. Expression achieves maximum values at 15 hpi, declining at subsequent time points. These results may suggest, therefore, that cutinase most likely plays a role in P. infestans pathogenicity.
Waugh, M; Hraber, P; Weller, J; Wu, Y; Chen, G; Inman, J; Kiphart, D; Sobral, B
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 http:www.ncgr.org/pgi, with database access through the database access hyperlink.
Zhang, Gao-sheng; Wang, Ren-qing; Song, Bai-min
Data were collected in different successional stages using a simultaneous sampling method and analyzed through quantitative classification method. Three large groups and 12 classes were made to represent the community patterns of three succession stages and 12 succession communities. The succession series of plant community in the study area was as follows: saline bare land→community Suaeda salsa→community Tamarix chinensis→grassland. Succession degree and succession process of 12 succession communities were calculated. Most of these communities were in the lower succession stage, however, community Phragmites communis+Glycine soja and community Imperata cylindrica+G. soja were close to the succession stage of grassland climax. Five species diversity indices were used to study the changes in species richness, species evenness and diversity during succession of community. Heterogeneity index and richness index increased gradually during the community succession process, but species evenness tended to decrease with succession development. The relation between succession and environment was studied by ordination technique, and the results showed that the soil salt content was an important factor to halarch succession of the modern Yellow River Delta. It affected community structure, species composition and succession process. PMID:17657854
Song, Tianqiao; Ma, Zhenchuan; Shen, Danyu; Li, Qi; Li, Wanlin; Su, Liming; Ye, Tingyue; Zhang, Meixiang; Wang, Yuanchao; Dou, Daolong
Oomycete pathogens produce a large number of CRN effectors to manipulate plant immune responses and promote infection. However, their functional mechanisms are largely unknown. Here, we identified a Phytophthora sojae CRN effector PsCRN108 which contains a putative DNA-binding helix-hairpin-helix (HhH) motif and acts in the plant cell nucleus. Silencing of the PsCRN108 gene reduced P. sojae virulence to soybean, while expression of the gene in Nicotiana benthamiana and Arabidopsis thaliana enhanced plant susceptibility to P. capsici. Moreover, PsCRN108 could inhibit expression of HSP genes in A. thaliana, N. benthamiana and soybean. Both the HhH motif and nuclear localization signal of this effector were required for its contribution to virulence and its suppression of HSP gene expression. Furthermore, we found that PsCRN108 targeted HSP promoters in an HSE- and HhH motif-dependent manner. PsCRN108 could inhibit the association of the HSE with the plant heat shock transcription factor AtHsfA1a, which initializes HSP gene expression in response to stress. Therefore, our data support a role for PsCRN108 as a nucleomodulin in down-regulating the expression of plant defense-related genes by directly targeting specific plant promoters. PMID:26714171
Wicklow, D. T.
Conidia of domesticated yellow-green aspergilli from strains of Aspergillus oryzae (Ahlburg) Cohn and Aspergillus sojae Sakaguchi and Yamada ex Murakami, used in the preparation of koji inoculum, germinate approximately 3 h sooner than conidia produced by related wild species, Aspergillus flavus Link ex Fr. and Aspergillus parasiticus Speare. There was no consistent relationship between average conidium size and estimated 50% germination time. Germination trials were conducted on Czapek agar at 28°C. A hypothesis is offered that, in the propagation of koji inoculum, selection has favored those individuals capable of rapid conidium germination and germ tube extension, attributes that enable them to gain the available substrate during intraspecific competition. PMID:16346471
Gutierrez, Kevin J; DasGupta, Sayantani
Feminist theorist and educator, bell hooks, asserts that to seek true liberation one must choose marginality. One must choose to occupy the space outside the binary between colonizer-colonized, hegemonic center-periphery, and us-them in order to create a location of possibility. This essay will reveal the practice of social justice as the navigation of the space that difference makes and argue that choosing marginality provides a framework for health humanities work towards social justice in health care. The space of the launderette that is depicted in Hanif Kureishi's 1986 film, My Beautiful Laundrette, provides an example of choosing marginality and illustrates how difference structures both real and imagined spaces, which influences how individuals ultimately perceive one another. We will draw from the work of bell hooks; political geographer, Edward Soja; and Marxist philosopher, Henri Lefebvre, to demonstrate the importance of the health humanities' position at the margin to traditional health care education.
Aparicio, V A; Sánchez, C; Ortega, F B; Nebot, E; Kapravelou, G; Porres, J M; Aranda, P
Introducción: La cantidad y la fuente de proteína, el entrenamiento de fuerza hipertrofia (EF) y los esteroides anabolizantes androgénicos (EAA) pueden alterar el peso corporal y el perfil lipídico plasmático y hepático. Material y métodos: 157 ratas Wistar adultas macho se distribuyeron al azar en 16 grupos experimentales del siguiente modo: dietas normoproteica (NP) o hiperproteicas (HP), proteínas de lactosuero o de soja, con y sin EF y con o sin EAA, durante un periodo experimental de 3 meses. Resultados y discusión: El peso corporal final fue menor en los grupos con EF y EAA en comparación con los grupo sedentario y sin EAA, respectivamente (todos, p<0,001). El colesterol plasmático total (CT) fue menor en el grupo con dieta HP en comparación con las dieta NP, para las dietas de proteínas de lactosuero en comparación con las proteínas de soja, y para el grupo con EAA en comparación con el grupo sin EAA (todos, p<0,001). Las concentraciones plasmáticas de colesterol HDL fueron superiores en los grupos de EF (p<0,05) y menores en los grupos con EAA (p<0,001), y de dieta HP o con proteína de soja (p<0,05). Los triglicéridos (TAG) plasmáticos fueron menores con la dieta HP (p<0,001), el EF (p=0,002) y la no administración de EAA (p=0,001). El CT hepático fue menor en los grupos de dieta NP (p<0,01), dieta con proteínas de soja (p<0,05) y grupo de EAA (p<0,001). Los TAG hepáticos fueron menores en los grupos de dieta de proteínas de lactosuero (p<0,001), EF y sin EAA (ambos, p<0,05). Se hallaron algunas interacciones como un mayor efecto de los EAA en la reducción del peso corporal de las ratas que realizaron EF o ingirieron una dieta HP (todos, p<0,05). Las concentraciones plasmáticas de colesterol HDL fueron superiores cuando se combinó el EF con las dietas HP (p=0,010) o sin EAA y cuando las dietas HP se combinaron con el no uso de EAA (ambos, p<0,001). Finalmente, los grupos que combinaron el EF sin EAA obtuvieron los valores m
Kamoun, Sophien; Furzer, Oliver; Jones, Jonathan D G; Judelson, Howard S; Ali, Gul Shad; Dalio, Ronaldo J D; Roy, Sanjoy Guha; Schena, Leonardo; Zambounis, Antonios; Panabières, Franck; Cahill, David; Ruocco, Michelina; Figueiredo, Andreia; Chen, Xiao-Ren; Hulvey, Jon; Stam, Remco; Lamour, Kurt; Gijzen, Mark; Tyler, Brett M; Grünwald, Niklaus J; Mukhtar, M Shahid; Tomé, Daniel F A; Tör, Mahmut; Van Den Ackerveken, Guido; McDowell, John; Daayf, Fouad; Fry, William E; Lindqvist-Kreuze, Hannele; Meijer, Harold J G; Petre, Benjamin; Ristaino, Jean; Yoshida, Kentaro; Birch, Paul R J; Govers, Francine
Oomycetes form a deep lineage of eukaryotic organisms that includes a large number of plant pathogens which threaten natural and managed ecosystems. We undertook a survey to query the community for their ranking of plant-pathogenic oomycete species based on scientific and economic importance. In total, we received 263 votes from 62 scientists in 15 countries for a total of 33 species. The Top 10 species and their ranking are: (1) Phytophthora infestans; (2, tied) Hyaloperonospora arabidopsidis; (2, tied) Phytophthora ramorum; (4) Phytophthora sojae; (5) Phytophthora capsici; (6) Plasmopara viticola; (7) Phytophthora cinnamomi; (8, tied) Phytophthora parasitica; (8, tied) Pythium ultimum; and (10) Albugo candida. This article provides an introduction to these 10 taxa and a snapshot of current research. We hope that the list will serve as a benchmark for future trends in oomycete research.
Silva Ton, Winder Tadeu; das Graças de Almeida, Crislaine; de Morais Cardoso, Leandro; Marvila Girondoli, Yassana; Feliciano Pereira, Patrícia; Viana Gomes Schitini, Josiane Keila; Galvão Cândido, Flávia; Marques Arbex, Priscila; de Cássia Gonçalves Alfenas, Rita
Introducción: La diabetes mellitus es una enfermedad epidémica que afecta a 346 millones de personas en el mundo. El control glicémico es la clave para la prevención y el control de la diabetes. Algunas proteínas pueden estimular la liberación de insulina y modular la respuesta glicémica. Objetivos: Evaluar el efecto del consumo de diferentes tipos de proteínas (proteína de suero de leche, proteína de soja y la clara de huevo) de la glicemia postprandial en una segunda comida en individuos de peso normal y normoglicémicos. Metodología: Este fue un ensayo clínico aleatorizado cruzado. Después de un ayuno nocturno de 12 horas, diez individuos asistieron al laboratorio para beber uno de los batidos de proteínas (suero de leche, soja o clara de huevo) o la bebida control. Treinta minutos más tarde, los individuos consumieron una solución de glucosa (25 g de glucosa). La respuesta glicémica fue monitorizada en los tiempos 0 (antes de solución de glucosa) y 15, 30, 45, 60, 90 y 120 min (después del consumo de la solución de glucosa). El área incrementada bajo la curva de glicemia (iAUC) fue calculada por el método trapezoidal. Por otra parte, la respuesta glicémica se evaluó mediante un nuevo método que utiliza la ecuación de iG. Resultados: En comparación con el control, las bebidas de suero de leche y de proteína de soja reducen iAUC postprandial en 56,5% (p = 0,004) y 44,4% (p = 0,029), respectivamente. La proteína de suero es la única proteína capaz de evitar grandes fluctuaciones y un pico de glicemia postprandial. La evaluación de la respuesta glicémica por la ecuación iG mostró correlación positiva con iAUC (Pearson 0,985, p < 0,05). Conclusión: El consumo de suero de leche y proteína de soja 30 minutos antes de una carga de glucosa resultó en menor iAUC en comparación con la bebida control. La proteína del suero mantiene la glucemia postprandial más estable.
Norman, Julian; Groisman, Pavel; Soja, Amber J.
In 2007 and 2009 Environmental Research Letters published focus issues (edited by Pavel Groisman and Amber J Soja) made up of work carried out by NEESPI participants. Here, we present the content of those focus issues as an invaluable resource for researchers working in the NEESPI study area. The first of the two issues, published in 2007 with title 'Northern Hemisphere High Latitude Climate and Environmental Change', presents a diverse collection of articles that are assembled into five groups devoted to studies of climate and hydrology, land cover and land use, the biogeochemical cycle and its feedbacks, the cryosphere, and human dimensions. The second issue, published in 2009, with title 'Climatic and Environmental Change in Northern Eurasia' presents diverse, assorted studies of different aspects of contemporary change, representing the diversity of climates and ecosystems across Northern Eurasia.
Heinkelmann, R.; Soja, B.; Schuh, H.
In 2011 and 2012 the IVS observed twelve VLBI research and development (R&D) sessions that include successful observations as angularly close as 3.9° from the heliocenter. Among others, one purpose of these IVS-R&D sessions was to achieve an improvement in the determination of the PPN parameter γ . Besides, by analyzing this specific set of IVS sessions, it was for the first time possible to measure the dispersive effect of the Solar corona with VLBI (Soja et al., 2014). In this work we assess the formal error of the γ-parameter and the contributions of the various terms to the partial derivative of the γ-parameter. Furthermore, we investigate the size of the gravitational delays caused by: (i) Solar monopole field at rest and with approximately linear translation, (ii) rotation of the Solar monopole field, (iii) Solar gravitational field quadrupole expansion, and (iv) Solar higher order term.
Nierle, W; el Wahab el Bayâ, A
The chemical composition of some legume seeds is reviewed. It was found that soy (Glycine soja), field beans (Vicia faba), Vicia sativa and Lens culingris (red seeds) have the highest protein content. There were only minor differences in amino acid composition except for glutamic acid. The lysine content of legume seeds was high. Lysine is the most limiting essential amino acid in cereal proteins, therefore cereal products can be improved nutritionally by addition of legume seed flour. Examination of polar lipids by thin layer chromatography revealed that lecithin, phosphatidyl ethanolamine and phosphatidyl inositol were the major components. However, Lupinus angustifolius contained two unknown polar lipid classes.--In contrary to previous literature the ripe legume seeds contained only a very small amount of galactosyl diglycerides; however legume seeds contain higher amounts of the compounds. The conection between biosynthesis of linolenic acid and the presence of galactosyl diglycerides in higher plants is discussed.
Ortiz, Gastón Ezequiel; Noseda, Diego Gabriel; Ponce Mora, María Clara; Recupero, Matías Nicolás; Blasco, Martín; Albertó, Edgardo
A comparative study of the proteolytic enzymes production using twelve Aspergillus strains previously unused for this purpose was performed by solid state fermentation. A semiquantitative and quantitative evaluation of proteolytic activity were carried out using crude enzymatic extracts obtained from the fermentation cultures, finding seven strains with high and intermediate level of protease activity. Biochemical, thermodynamics, and kinetics features such as optimum pH and temperature values, thermal stability, activation energy (Ea), quotient energy (Q10), Km, and Vmax were studied in four enzymatic extracts from the selected strains that showed the highest productivity. Additionally, these strains were evaluated by zymogram analysis obtaining protease profiles with a wide range of molecular weight for each sample. From these four strains with the highest productivity, the proteolytic extract of A. sojae ATCC 20235 was shown to be an appropriate biocatalyst for hydrolysis of casein and gelatin substrates, increasing its antioxidant activities in 35% and 125%, respectively. PMID:26989505
Qiao, Yongli; Liu, Lin; Xiong, Qin; Flores, Cristina; Wong, James; Shi, Jinxia; Wang, Xianbing; Liu, Xigang; Xiang, Qijun; Jiang, Shushu; Zhang, Fuchun; Wang, Yuanchao; Judelson, Howard S; Chen, Xuemei; Ma, Wenbo
Effectors are essential virulence proteins produced by a broad range of parasites, including viruses, bacteria, fungi, oomycetes, protozoa, insects and nematodes. Upon entry into host cells, pathogen effectors manipulate specific physiological processes or signaling pathways to subvert host immunity. Most effectors, especially those of eukaryotic pathogens, remain functionally uncharacterized. Here, we show that two effectors from the oomycete plant pathogen Phytophthora sojae suppress RNA silencing in plants by inhibiting the biogenesis of small RNAs. Ectopic expression of these Phytophthora suppressors of RNA silencing enhances plant susceptibility to both a virus and Phytophthora, showing that some eukaryotic pathogens have evolved virulence proteins that target host RNA silencing processes to promote infection. These findings identify RNA silencing suppression as a common strategy used by pathogens across kingdoms to cause disease and are consistent with RNA silencing having key roles in host defense.
de Freitas Folly, Gilce Andrezza; Neiva da Silva, Ester; Vieira Verner, Fabiana; Dos Santos Silva, Fernanda Cacilda; Pinheiro Volp, Ana Carolina
Introducción: Los prebióticos y alimentos con propiedades funcionales proporcionan beneficios para la salud de los consumidores a través de la prevención de muchas enfermedades. Objetivo: Verificar la aceptación de productos artesanales (chocolate en barra, dulce de soja y pan dulce) formulados con nueces (linaza, maní y nueces de Brasil) y, o fructooligosacáridos (FOS). Métodos: Cuatro muestras de cada producto fueron preparados con adición de diferentes concentraciones de nueces y FOS. La evaluación sensorial se realizó mediante una muestra de 373 consumidores, con 126, 121 y 126 probadores para muestras de chocolate en barra, dulce de soja y pan dulce, respectivamente, utilizándose la escala hedónica de nueve puntos. Los resultados fueron sometidos a Análisis de Varianza (ANOVA) y el test de Tukey. Resultados y Discusión: Observándose las medias de los juzgamientos, se infiere que las muestras con mayor aceptación han sido de pan dulce con nueces de Brasil y, o FOS. Sin embargo, todas las muestras son buenas alternativas de mercado y se lo mostró un promedio de entre 6 y 9 puntos, más un aumento de ácidos grasos omega 3 y 6, proteínas, fibras, vitaminas, antioxidantes y minerales, así como fitoquímicos, los cuales desempeñan un papel importante en la promoción de la salud. Conclusión: Los productos artesanales formulados con oleaginosas y, o FOS tuvieron una buena aceptabilidad y pueden mejorar los hábitos alimentarios de los consumidores. Pero para probar la funcionalidad de estos productos, se necesitan nuevos estudios.
Vieira de Melo, Ingrid Sofia; Da Rocha Ataide, Terezinha; Lima de Oliveira, Suzana; Bezerra Bueno, Nassib; Duarte de Freitas, Johnnatan; Goulart Sant'Ana, Antônio Euzébio
Objetivo: el objetivo de este estudio fue evaluar la influencia del consumo de una dieta cetogénica complementada con triheptanoína, un triacilglicerol de cadena media y anaplerótico, en el perfil de ácidos grasos del hígado de ratones Wistar. Métodos: tres grupos de ratones Wistar machos (n = 10) fueron sometidos durante 60 días a una dieta AIN-93 de control, una dieta cetogénica basada en triheptanoína o una dieta cetogénica a base de aceite de soja. Los hígados fueron escindidos y sometidos a extracción de lípidos y metilación para obtener los ésteres metílicos de ácidos grasos, que se sometieron a cromatografía de gas-espectrometría de masa. Resultados y discusión: en comparación con los ratones alimentados con la dieta de control, los de ambas dietas cetogénicas mostraron una reducción significativa en las concentraciones de los ácidos grasos 9-hexadecenoico y 9-octadecenoico, mientras que los alimentados con triheptanoína mostraron niveles de ácido octadecenoico aumentados. Conclusión: los cambios en los perfiles de ácidos grasos del hígado de los ratones alimentados con dietas cetogénicas no están relacionados con la fuente de grasa de la dieta (triheptanoína o aceite de soja), sino más bien con la concentración total de lípidos.
Guo, Hui Juan; Wang, En Tao; Zhang, Xing Xing; Li, Qin Qin; Zhang, Yan Ming; Tian, Chang Fu; Chen, Wen Xin
In order to investigate the genetic differentiation of Sinorhizobium strains nodulating Glycine max and related microevolutionary mechanisms, three housekeeping genes (SMc00019, truA, and thrA) and 16 symbiosis-related genes on the chromosome (7 genes), pSymA (6 genes), and pSymB (3 genes) were analyzed. Five distinct species were identified among the test strains by calculating the average nucleotide identity (ANI) of SMc00019-truA-thrA: Sinorhizobium fredii, Sinorhizobium sojae, Sinorhizobium sp. I, Sinorhizobium sp. II, and Sinorhizobium sp. III. These species assignments were also supported by population genetics and phylogenetic analyses of housekeeping genes and symbiosis-related genes on the chromosome and pSymB. Different levels of genetic differentiation were observed among these species or different replicons. S. sojae was the most divergent from the other test species and was characterized by its low intraspecies diversity and limited geographic distribution. Intergenic recombination dominated the evolution of 19 genes from different replicons. Intraspecies recombination happened frequently in housekeeping genes and symbiosis-related genes on the chromosome and pSymB, whereas pSymA genes showed a clear pattern of lateral-transfer events between different species. Moreover, pSymA genes were characterized by a lower level of polymorphism and recombination than those on the chromosome and pSymB. Taken together, genes from different replicons of rhizobia might be involved in the establishment of symbiosis with legumes, but these symbiosis-related genes might have evolved differently according to their corresponding replicons.
Xu, Zhen; Jiang, Huawei; Sahu, Binod Bihari; Kambakam, Sekhar; Singh, Prashant; Wang, Xinran; Wang, Qiugu; Bhattacharyya, Madan K; Dong, Liang
This paper reports a highly economical and accessible approach to generate different discrete relative humidity conditions in spatially separated wells of a modified multi-well plate for humidity assay of plant-pathogen interactions with good throughput. We demonstrated that a discrete humidity gradient could be formed within a few minutes and maintained over a period of a few days inside the device. The device consisted of a freeway channel in the top layer, multiple compartmented wells in the bottom layer, a water source, and a drying agent source. The combinational effects of evaporation, diffusion, and convection were synergized to establish the stable discrete humidity gradient. The device was employed to study visible and molecular disease phenotypes of soybean in responses to infection by Phytophthora sojae, an oomycete pathogen, under a set of humidity conditions, with two near-isogenic soybean lines, Williams and Williams 82, that differ for a Phytophthora resistance gene (Rps1-k). Our result showed that at 63% relative humidity, the transcript level of the defense gene GmPR1 was at minimum in the susceptible soybean line Williams and at maximal level in the resistant line Williams 82 following P. sojae CC5C infection. In addition, we investigated the effects of environmental temperature, dimensional and geometrical parameters, and other configurational factors on the ability of the device to generate miniature humidity environments. This work represents an exploratory effort to economically and efficiently manipulate humidity environments in a space-limited device and shows a great potential to facilitate humidity assay of plant seed germination and development, pathogen growth, and plant-pathogen interactions. Since the proposed device can be easily made, modified, and operated, it is believed that this present humidity manipulation technology will benefit many laboratories in the area of seed science, plant pathology, and plant-microbe biology, where
Belbahri, Lassaad; Calmin, Gautier; Mauch, Felix; Andersson, Jan O
Lateral gene transfer (LGT) can facilitate the acquisition of new functions in recipient lineages, which may enable them to colonize new environments. Several recent publications have shown that gene transfer between prokaryotes and eukaryotes occurs with appreciable frequency. Here we present a study of interdomain gene transfer of cutinases -- well documented virulence factors in fungi -- between eukaryotic plant pathogens Phytophthora species and prokaryotic bacterial lineages. Two putative cutinase genes were cloned from Phytophthora brassicae and Northern blotting experiments showed that these genes are expressed early during the infection of the host Arabidopsis thaliana and induced during cyst germination of the pathogen. Analysis of the gene organisation of this gene family in Phytophthora ramorum and P. sojae showed three and ten copies in tight succession within a region of 5 and 25 kb, respectively, probably indicating a recent expansion in Phytophthora lineages by gene duplications. Bioinformatic analyses identified orthologues only in three genera of Actinobacteria, and in two distantly related eukaryotic groups: oomycetes and fungi. Together with phylogenetic analyses this limited distribution of the gene in the tree of life strongly support a scenario where cutinase genes originated after the origin of land plants in a microbial lineage living in proximity of plants and subsequently were transferred between distantly related plant-degrading microbes. More precisely, a cutinase gene was likely acquired by an ancestor of P. brassicae, P. sojae, P. infestans and P. ramorum, possibly from an actinobacterial source, suggesting that gene transfer might be an important mechanism in the evolution of their virulence. These findings could indeed provide an interesting model system to study acquisition of virulence factors in these important plant pathogens.
Bollmann, Stephanie R.; Fang, Yufeng; Press, Caroline M.; Tyler, Brett M.; Grünwald, Niklaus J.
Gene regulation by small RNA pathways is ubiquitous among eukaryotes, but little is known about small RNA pathways in the Stramenopile kingdom. Phytophthora, a genus of filamentous oomycetes, contains many devastating plant pathogens, causing multibillion-dollar damage to crops, ornamental plants, and natural environments. The genomes of several oomycetes including Phytophthora species such as the soybean pathogen P. sojae, have been sequenced, allowing evolutionary analysis of small RNA-processing enzymes. This study examined the evolutionary origins of the oomycete small RNA-related genes Dicer-like (DCL), and RNA-dependent RNA polymerase (RDR) through broad phylogenetic analyses of the key domains. Two Dicer gene homologs, DCL1 and DCL2, and one RDR homolog were cloned and analyzed from P. sojae. Gene expression analysis revealed only minor changes in transcript levels among different life stages. Oomycete DCL1 homologs clustered with animal and plant Dicer homologs in evolutionary trees, whereas oomycete DCL2 homologs clustered basally to the tree along with Drosha homologs. Phylogenetic analysis of the RDR homologs confirmed a previous study that suggested the last common eukaryote ancestor possessed three RDR homologs, which were selectively retained or lost in later lineages. Our analysis clarifies the position of some Unikont and Chromalveolate RDR lineages within the tree, including oomycete homologs. Finally, we analyzed alterations in the domain structure of oomycete Dicer and RDR homologs, specifically focusing on the proposed domain transfer of the DEAD-box helicase domain from Dicer to RDR. Implications of the oomycete domain structure are discussed, and possible roles of the two oomycete Dicer homologs are proposed. PMID:27014308
Liu, Li-hua; Jiang, Jing-yan; Zong, Liang-gang
Burning of agricultural crop residues was a major source greenhouse gases. In this study, the proportion of crop straws (rice, wheat, maize, oil rape, cotton and soja) in Jiangsu used as household fuel and direct open burning in different periods (1990-1995, 1996-2000, 2001-2005 and 2006-2008) was estimated through questionnaire. The emission factors of CO2, CO, CH4 and NO20 from the above six types of crop straws were calculated by the simulated burning experiment. Thus the emission inventory of greenhouse gases from crop straws burning was established according to above the burning percentages and emission factors, ratios of dry residues to production and crop productions of different periods in Jiangsu province. Results indicated that emission factors of CO2, CO, CH4 and N2O depended on crop straw type. The emission factors of CO2 and CH4 were higher for oil rape straw than the other straws, while the maize and the rice straw had the higher N2O and CO emission factor. Emission inventory of greenhouse gases from agricultural residues burning in Jiangsu province showed, the annual average global warming potential (GWP) of six tested crop straws were estimated to be 9.18 (rice straw), 4.35 (wheat straw), 2.55 (maize straw), 1.63 (oil rape straw), 0.55 (cotton straw) and 0. 39 (soja straw) Tg CO2 equivalent, respectively. Among the four study periods, the annual average GWP had no obvious difference between the 1990-1995 and 2006-2008 periods, while the maximal annual average GWP (23.83 Tg CO2 equivalent) happened in the 1996-2000 period, and the minimum (20.30 Tg CO2 equivalent) in 1996-2000 period.
Garib, Ricardo; Garla, Priscila; Torrinhas, Raquel S; Bertevello, Pedro L; Logullo, Angela F; Waitzberg, Dan L
Objetivo: Estudiar los efectos de los diferentes protocolos de infusión de la emulsion de lípidos de aceite de pescado (Fole) sobre la inflamación aguda en el modelo de colitis en la rata. Material y métodos: Ratas Wistar macho adultas (n = 51) fueron asignados al azar en 5 grupos para recibir infusión parenteral de solución salina (SS) o emulsión de lípidos de aceite de soja (SO), como controles, y Fole compone de: aceite de pescado solo (FO), una mezcla (9:1 v/v) de SO con FO (SO/FO), o 30% de aceite de soja, 30% triglicéridos de cadena media, 25% de aceite de oliva, y 15% de aceite de pescado (SMOF). Después de 72 h de infusión intravenosa, colitis experimental fue inducida con ácido acético. Después de 24 h, las muestras de colon se analizaron para determinar cambios histológicos y citoquinas. Resultados: En relación en el SS grupo, necrosis macroscópica fue menos frecuente en el grupo FO y necrosis histológica fue más frecuente en el grupo de SMOF. Existe una relación directa e inversa de colon interleuquina (IL) -1 e IL-4, respectivamente, con necrosis histológica. En comparación con el grupo SS, en el FO hubo aumento de IL-4 e IFN-gamma y disminución de TNF-alfa, SO/FO disminuyó TNF-alfa, y en el SMOF hubo aumento de IL-1 y la disminución de IL-4. Conclusión: En la colitis inducida por ácido acético, la infusion aislada de Fole compuesto de aceite de pescado por sí solo fue más ventajosa en la atenuacion de la inflamacióndo que la infusión de Fole contiendo otros aceites, y esta diferencia puede ser debida las influencias de su diferente contenido de ácido graso.
Michczynska, D. J.; Hajdas, I.
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: Michczyńska 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. Michczyńska D.J., Michczyński 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., Michczyńska 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., Michczyńska D.J., 2006. Past hydrological events reflected in
Adak, Totan; Kumar, Jitendra; Dey, Debjani; Shakil, N A; Walia, S
Controlled release (CR) formulations of imidacloprid (1-(6 chloro-3-pyridinyl methyl)-N- nitro imidazolidin-2- ylideneamine) were prepared using novel amphiphilic polymers synthesized from polyethylene glycol and aliphatic diacids employing encapsulation technique. The bioefficacy of the prepared CR formulations was evaluated against major pests of soybean, namely stem fly, Melanagromyza sojae Zehntmer and white fly, Bemisia tabaci Gennadius along with a commercial formulation at the experimental farm of Indian Agricultural Research Institute (IARI), New Delhi during kharif 2009 and 2010. Most of the CR formulations of imidacloprid gave significantly better control of the pests compare to its commercial formulations, however the CR formulations, Poly [poly (oxyethylene-1000)-oxy suberoyl] amphiphilic polymer based formulation performed better over others for controlling of both stem fly incidence and Yellow Mosaic Virus (YMV) infestation transmitted by white fly. Some of the developed CR formulations recorded higher yield over commercial formulation and control. Nodulation pattern of soybean was not affected due to treatment of CR and commercial formulations of imidacloprid. Also the residues of imidacloprid in seed and soil at harvest were not detectable for both CR and commercial formulations.
Yang, Bo; Wang, Qunqing; Jing, Maofeng; Guo, Baodian; Wu, Jiawei; Wang, Haonan; Wang, Yang; Lin, Long; Wang, Yan; Ye, Wenwu; Dong, Suomeng; Wang, Yuanchao
Phytophthora pathogens secrete effectors to manipulate host innate immunity, thus facilitating infection. Among the RXLR effectors highly induced during Phytophthora sojae infection, Avh238 not only contributes to pathogen virulence but also triggers plant cell death. However, the detailed molecular basis of Avh238 functions remains largely unknown. We mapped the regions responsible for Avh238 functions in pathogen virulence and plant cell death induction using a strategy that combines investigation of natural variation and large-scale mutagenesis assays. The correlation between cellular localization and Avh238 functions was also evaluated. We found that the 79(th) residue (histidine or leucine) of Avh238 determined its cell death-inducing activity, and that the 53 amino acids in its C-terminal region are responsible for promoting Phytophthora infection. Transient expression of Avh238 in Nicotiana benthamiana revealed that nuclear localization is essential for triggering cell death, while Avh238-mediated suppression of INF1-triggered cell death requires cytoplasmic localization. Our results demonstrate that a representative example of an essential Phytophthora RXLR effector can evolve to escape recognition by the host by mutating one nucleotide site, and can also retain plant immunosuppressive activity to enhance pathogen virulence in planta.
Lin, Sen; Li, Qing; Yang, Bao; Duan, Xuewu; Zhang, Mingwei; Shi, John; Jiang, Yueming
Condensed tannin is a ubiquitous polyphenol in plants that possesses substantial antioxidant capacity. In this study, we have investigated the polyphenol extraction recovery and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity of the extracted polyphenol after litchi pericarp is treated with Aspergillus awamori, Aspergillus sojae or Aspergillus oryzae. We have further explored the activity of A. awamori in the formation of condensed tannin. The treatment of A. awamori appeared to produce the highest antioxidant activity of polyphenol from litchi pericarp. Further studies suggested that the treatment of A. awamori releases the non-extractable condensed tannin from cell walls of litchi pericarp. The total extractable tannin in the litchi pericarp residue after a six-time extraction with 60% ethanol increased from 199.92 ± 14.47-318.38 ± 7.59 μg/g dry weight (DW) after the treatment of A. awamori. The ESI-TOF-MS and HPLC-MS² analyses further revealed that treatment of A. awamori degraded B-type condensed tannin (condensed flavan-3-ol via C4-C8 linkage), but exhibited a limited capacity to degrade the condensed tannin containing A-type linkage subunits (C4-C8 coupled C2-O-C7 linkage). These results suggest that the treatment of A. awamori can significantly improve the production of condensed tannin from litchi pericarp.
Kong, Ping; Hong, Chuanxue
Phytophthora species are known as “plant destroyers” capable of initiating single zoospore infection in the presence of a quorum of chemical signals from the same or closely related species of oomycetes. Since the natural oomycete population is too low to reach a quorum necessary to initiate a disease epidemic, creation of the quorum is reliant on alternate sources. Here, we show that a soil bacterial isolate, Bacillus megaterium Sb5, promotes plant infection by Phytophthora species. In the presence of Sb5 exudates, colonization of rhododendron leaf discs by 12 Phytophthora species/isolates was significantly enhanced, single zoospores of P. nicotianae infected annual vinca and P. sojae race 25 successfully attacked a non-host plant, Nicotiana benthamiana as well as resistant soybean cultivars with RPS1a or RPS3a. Sb5 exudates, most notably the fractions larger than 3 kDa, promoted plant infection by improving zoospore swimming, germination and plant attachment. Sb5 exudates also stimulated infection hypha growth and upregulated effector gene expression. These results suggest that environmental bacteria are important sources of virulence signal providers that promote plant infection by Phytophthora species, advancing our understanding of biotic factors in the environmental component of the Phytophthora disease triangle and of communal infection of plant pathogens. PMID:27616267
Brouwer, Henk; Coutinho, Pedro M; Henrissat, Bernard; de Vries, Ronald P
Carbohydrate-Active enZymes (CAZymes) form particularly interesting targets to study in plant pathogens. Despite the fact that many CAZymes are pathogenicity factors, oomycete CAZymes have received significantly less attention than effectors in the literature. Here we present an analysis of the CAZymes present in the Phytophthora infestans, Ph. ramorum, Ph. sojae and Pythium ultimum genomes compared to growth of these species on a range of different carbon sources. Growth on these carbon sources indicates that the size of enzyme families involved in degradation of cell-wall related substrates like cellulose, xylan and pectin is not always a good predictor of growth on these substrates. While a capacity to degrade xylan and cellulose exists the products are not fully saccharified and used as a carbon source. The Phytophthora genomes encode larger CAZyme sets when compared to Py. ultimum, and encode putative cutinases, GH12 xyloglucanases and GH10 xylanases that are missing in the Py. ultimum genome. Phytophthora spp. also encode a larger number of enzyme families and genes involved in pectin degradation. No loss or gain of complete enzyme families was found between the Phytophthora genomes, but there are some marked differences in the size of some enzyme families.
Guan, Zheng-Jun; Zhang, Peng-Fei; Wei, Wei; Mi, Xiang-Cheng; Kang, Ding-Ming; Liu, Biao
Gene flow from genetically modified (GM) crops to wild relatives might affect the evolutionary dynamics of weedy populations and result in the persistence of escaped genes. To examine the effects of this gene flow, the growth of F1 hybrids that were formed by pollinating wild soybean (Glycine soja) with glyphosate-tolerant GM soybean (G. max) or its non-GM counterpart was examined in a greenhouse. The wild soybean was collected from two geographical populations in China. The performance of the wild soybean and the F2 hybrids was further explored in a field trial. Performance was measured by several vegetative and reproductive growth parameters, including the vegetative growth period, pod number, seed number, above-ground biomass and 100-seed weight. The pod setting percentage was very low in the hybrid plants. Genetically modified hybrid F1 plants had a significantly longer period of vegetative growth, higher biomass and lower 100-seed weight than the non-GM ones. The 100-seed weight of both F1 and F2 hybrids was significantly higher than that of wild soybean in both the greenhouse and the field trial. No difference in plant growth was found between GM and non-GM F2 hybrids in the field trial. The herbicide-resistant gene appeared not to adversely affect the growth of introgressed wild soybeans, suggesting that the escaped transgene could persist in nature in the absence of herbicide use.
Zhang, Jing; Yang, Dongshuang; Li, Mingxia; Shi, Lianxuan
Clarification of the metabolic mechanisms underlying salt stress responses in plants will allow further optimization of crop breeding and cultivation to obtain high yields in saline-alkali land. Here, we characterized 68 differential metabolites of cultivated soybean (Glycine max) and wild soybean (Glycine soja) under neutral-salt and alkali-salt stresses using gas chromatography-mass spectrometry (GC-MS)-based metabolomics, to reveal the physiological and molecular differences in salt tolerance. According to comparisons of growth parameters under the two kinds of salt stresses, the level of inhibition in wild soybean was lower than in cultivated soybean, especially under alkali-salt stress. Moreover, wild soybean contained significantly higher amounts of phenylalanine, asparagine, citraconic acid, citramalic acid, citric acid and α-ketoglutaric acid under neutral-salt stress, and higher amounts of palmitic acid, lignoceric acid, glucose, citric acid and α-ketoglutaric acid under alkali-salt stress, than cultivated soybean. Further investigations demonstrated that the ability of wild soybean to salt tolerance was mainly based on the synthesis of organic and amino acids, and the more active tricarboxylic acid cycle under neutral-salt stress. In addition, the metabolite profiling analysis suggested that the energy generation from β-oxidation, glycolysis and the citric acid cycle plays important roles under alkali-salt stress. Our results extend the understanding of mechanisms involved in wild soybean salt tolerance and provide an important reference for increasing yields and developing salt-tolerant soybean cultivars. PMID:27442489
Piotrowski, Jeff S.; Okada, Hiroki; Lu, Fachuang; ...
A rise in resistance to current antifungals necessitates strategies to identify alternative sources of effective fungicides. We report the discovery of poacic acid, a potent antifungal compound found in lignocellulosic hydrolysates of grasses. Chemical genomics using Saccharomyces cerevisiae showed that loss of cell wall synthesis and maintenance genes conferred increased sensitivity to poacic acid. Morphological analysis revealed that cells treated with poacic acid behaved similarly to cells treated with other cell wall-targeting drugs and mutants with deletions in genes involved in processes related to cell wall biogenesis. Poacic acid causes rapid cell lysis and is synergistic with caspofungin and fluconazole.more » The cellular target was identified; poacic acid localized to the cell wall and inhibited β-1,3-glucan synthesis in vivo and in vitro, apparently by directly binding β-1,3-glucan. Through its activity on the glucan layer, poacic acid inhibits growth of the fungi Sclerotinia sclerotiorum and Alternaria solani as well as the oomycete Phytophthora sojae. A single application of poacic acid to leaves infected with the broad-range fungal pathogen S. sclerotiorum substantially reduced lesion development. In conclusion, the discovery of poacic acid as a natural antifungal agent targeting β-1,3-glucan highlights the potential side use of products generated in the processing of renewable biomass toward biofuels as a source of valuable bioactive compounds and further clarifies the nature and mechanism of fermentation inhibitors found in lignocellulosic hydrolysates.« less
Zhou, Yuan; Yang, Yan; Zhou, Xinjian; Chi, Yingjun; Fan, Baofang; Chen, Zhixiang
Proteins containing the FxxxVQxhTG or VQ motif interact with WRKY transcription factors. Although VQ proteins have been reported in several plants, knowledge about their structures, functions and evolution is still very limited. Here, we report structural and functional analysis of the VQ protein family from soybean. Like Arabidopsis homologues, soybean VQ proteins bind only Group I and IIc WRKY proteins and a substantial number of their genes are responsive to stress-associated phytohormones. Overexpression of some soybean VQ genes in Arabidopsis had strong effects on plant growth, development, disease resistance and heat tolerance. Phylogenetic analysis, sequence alignment and site-directed mutagenesis revealed that the region immediately upstream of the FxxxVQxhTG motif also affects binding to WRKY proteins. Consistent with a larger WRKY-binding VQ domain, soybean VQ22 protein from cultivated soybean contains a 4-amino acid deletion in the region preceding its VQ motif that completely abolishes its binding to WRKY proteins. By contrast, the 4-amino acid deletion is absent in the VQ22 protein from wild soybean species (Glycine soja). Overexpression of wild soybean VQ22 in cultivated soybean inhibited growth, particularly after cold treatment. Thus, the mutation of soybean VQ22 is associated with advantageous phenotypes and may have been positively selected during evolution. PMID:27708406
Camiletti-Moirón, Daniel; Aparicio, Virginia Arianna; Nebot, Elena; Medina, Gerardo; Martínez, Rosario; Kapravelou, Garyfallia; Andrade, Ana; Porres, Jesús María; López-Jurado, María; Aranda, Pilar
Introducción: Es conocido que la proteína de soja así como la práctica de ejercicio físico aeróbico pueden incrementar la capacidad antioxidante y con ello reducir la sobreproducción de radicales libres en el cerebro. Sin embargo, existe desconocimiento sobre el efecto del consumo de dietas hiperproteicas y el entrenamiento de alta intensidad (EAI) sobre dicho estrés oxidativo. El objetivo del presente estudio fue analizar la influencia del consumo de una dieta hiperproteica y de EAI sobre marcadores de estrés oxidativo en cerebro. Métodos: Cuarenta ratas Wistar macho adultas fueron aleatoriamente distribuidas en 4 grupos experimentales (n=10): dieta normoproteica o hiperproteica, con o sin EAI durante un periodo experimental de 12 semanas. Se determinaron los principales marcadores de daño oxidativo en cerebro como sustancias reactivas del ácido tiobarbitúrico (TBARs) y el contenido de grupos carbonilos (PCC). Además, se midieron las actividades enzimáticas superóxido dismutasa del manganeso (Mn-SOD), de cobre/ zinc (CuZn-SOD) y catalasa (CAT), así como el nivel de proteína del factor nuclear eritroide-2 (Nrf2). Resultados: Los niveles de TBARs, PCC, tSOD, Mn- SOD, CuZn-SOD y CAT fueron significativamente mayores en los grupos hiperproteicos en comparación con los normoproteicos (todas, p.
Li, Kun-Lun; Bai, Xi; Li, Yong; Cai, Hua; Ji, Wei; Tang, Li-Li; Wen, Yi-Dong; Zhu, Yan-Ming
Low, but not freezing temperatures are a major factor limiting plant vegetative growth and development. It is not clear what signaling mechanism is used to limit plant growth in response to chronic low temperatures, thus it is important to continue to isolate and characterize genes whose activity/expression correlate well with cold responses. In this study, a novel GASA gene was isolated from Glycine soja and named GsGASA1. Quantitative real-time PCR analysis indicated that GsGASA1 expression responded to exogenous gibberellic acid (GA) and abscisic acid (ABA) treatments. Compared with wild-type plants under long-term cold treatment, the constitutive expression of GsGASA1 in transgenic Arabidopsis plants enhanced the inhibition of root elongation, while also increasing the transcript levels of RGL2 and RGL3, two of five DELLA genes in Arabidopsis. DELLA is a class of transcriptional regulators in GA signaling pathway restraining plant growth. Our results imply that GsGASA1 participates in chronic cold-induced root growth inhibition with the accumulation of DELLA genes. Lastly, a subcellular localization study using a yellow fluorescent protein (YFP) fusion protein indicated that GsGASA1 was localized to the plasma membrane, cytoplasm and nucleus.
León-Barrios, Milagros; Lorite, María José; Donate-Correa, Javier; Sanjuán, Juan
Eleven strains were isolated from root nodules of Lotus endemic to the Canary Islands and they belonged to the genus Ensifer, a genus never previously described as a symbiont of Lotus. According to their 16S rRNA and atpD gene sequences, two isolates represented minority genotypes that could belong to previously undescribed Ensifer species, but most of the isolates were classified within the species Ensifer meliloti. These isolates nodulated Lotus lancerottensis, Lotus corniculatus and Lotus japonicus, whereas Lotus tenuis and Lotus uliginosus were more restrictive hosts. However, effective nitrogen fixation only occurred with the endemic L. lancerottensis. The E. meliloti strains did not nodulate Medicago sativa, Medicago laciniata Glycine max or Glycine soja, but induced non-fixing nodules on Phaseolus vulgaris roots. nodC and nifH symbiotic gene phylogenies showed that the E. meliloti symbionts of Lotus markedly diverged from strains of Mesorhizobium loti, the usual symbionts of Lotus, as well as from the three biovars (bv. meliloti, bv. medicaginis, and bv. mediterranense) so far described within E. meliloti. Indeed, the nodC and nifH genes from the E. meliloti isolates from Lotus represented unique symbiotic genotypes. According to their symbiotic gene sequences and host range, the Lotus symbionts would represent a new biovar of E. meliloti for which bv. lancerottense is proposed.
Lorite, María J; Videira e Castro, Isabel; Muñoz, Socorro; Sanjuán, Juan
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.
Lin, Sen; Li, Qing; Yang, Bao; Duan, Xuewu; Zhang, Mingwei; Shi, John; Jiang, Yueming
Condensed tannin is a ubiquitous polyphenol in plants that possesses substantial antioxidant capacity. In this study, we have investigated the polyphenol extraction recovery and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity of the extracted polyphenol after litchi pericarp is treated with Aspergillus awamori, Aspergillus sojae or Aspergillus oryzae. We have further explored the activity of A. awamori in the formation of condensed tannin. The treatment of A. awamori appeared to produce the highest antioxidant activity of polyphenol from litchi pericarp. Further studies suggested that the treatment of A. awamori releases the non-extractable condensed tannin from cell walls of litchi pericarp. The total extractable tannin in the litchi pericarp residue after a six-time extraction with 60% ethanol increased from 199.92 ± 14.47–318.38 ± 7.59 μg/g dry weight (DW) after the treatment of A. awamori. The ESI-TOF-MS and HPLC-MS2 analyses further revealed that treatment of A. awamori degraded B-type condensed tannin (condensed flavan-3-ol via C4–C8 linkage), but exhibited a limited capacity to degrade the condensed tannin containing A-type linkage subunits (C4–C8 coupled C2–O–C7 linkage). These results suggest that the treatment of A. awamori can significantly improve the production of condensed tannin from litchi pericarp. PMID:27420043
Aspergillus is an important fungal genus used for the fermentation of Asian foods; this genus is referred to as koji mold in Japan and China. A. oryzae, A. sojae, and A. tamari are used in the production of miso and shoyu in Japan, but a comprehensive taxonomic study of Aspergillus isolated from Meju, a fermented soybean starting material for traditional soy sauce and soybean paste in Korea, has not been conducted. In this study, various Aspergillus species were isolated during a study of the mycobiota of Meju, and the aspergilli were identified based on phenotypic characteristics and sequencing of the β-tubulin gene. Most strains of Aspergillus were found to belong to the following sections: Aspergillus (n = 220), Flavi (n = 213), and Nigri (n = 54). The most commonly identified species were A. oryzae (n = 183), A. pseudoglaucus (Eurotium repens) (n = 81), A. chevalieri (E. chevalieri) (n = 62), A. montevidensis (E. amstelodami) (n = 34), A. niger (n = 21), A. tamari (n = 15), A. ruber (E. rubrum) (n = 15), A. proliferans (n = 14), and A. luchuensis (n = 14); 25 species were identified from 533 Aspergillus strains. Aspergillus strains were mainly found during the high temperature fermentation period in the later steps of Meju fermentation. PMID:26539037
Murphy, Guillermo P; Van Acker, Rene; Rajcan, Istvan; Swanton, Clarence J
Identity recognition systems allow plants to tailor competitive phenotypes in response to the genetic relatedness of neighbours. There is limited evidence for the existence of recognition systems in crop species and whether they operate at a level that would allow for identification of different degrees of relatedness. Here, we test the responses of commercial soya bean cultivars to neighbours of varying genetic relatedness consisting of other commercial cultivars (intraspecific), its wild progenitor Glycine soja, and another leguminous species Phaseolus vulgaris (interspecific). We found, for the first time to our knowledge, that a commercial soya bean cultivar, OAC Wallace, showed identity recognition responses to neighbours at different levels of genetic relatedness. OAC Wallace showed no response when grown with other commercial soya bean cultivars (intra-specific neighbours), showed increased allocation to leaves compared with stems with wild soya beans (highly related wild progenitor species), and increased allocation to leaves compared with stems and roots with white beans (interspecific neighbours). Wild soya bean also responded to identity recognition but these responses involved changes in biomass allocation towards stems instead of leaves suggesting that identity recognition responses are species-specific and consistent with the ecology of the species. In conclusion, elucidating identity recognition in crops may provide further knowledge into mechanisms of crop competition and the relationship between crop density and yield.
Guan, Zheng-Jun; Zhang, Peng-Fei; Wei, Wei; Mi, Xiang-Cheng; Kang, Ding-Ming; Liu, Biao
Gene flow from genetically modified (GM) crops to wild relatives might affect the evolutionary dynamics of weedy populations and result in the persistence of escaped genes. To examine the effects of this gene flow, the growth of F1 hybrids that were formed by pollinating wild soybean (Glycine soja) with glyphosate-tolerant GM soybean (G. max) or its non-GM counterpart was examined in a greenhouse. The wild soybean was collected from two geographical populations in China. The performance of the wild soybean and the F2 hybrids was further explored in a field trial. Performance was measured by several vegetative and reproductive growth parameters, including the vegetative growth period, pod number, seed number, above-ground biomass and 100-seed weight. The pod setting percentage was very low in the hybrid plants. Genetically modified hybrid F1 plants had a significantly longer period of vegetative growth, higher biomass and lower 100-seed weight than the non-GM ones. The 100-seed weight of both F1 and F2 hybrids was significantly higher than that of wild soybean in both the greenhouse and the field trial. No difference in plant growth was found between GM and non-GM F2 hybrids in the field trial. The herbicide-resistant gene appeared not to adversely affect the growth of introgressed wild soybeans, suggesting that the escaped transgene could persist in nature in the absence of herbicide use. PMID:26507568
Musket, Theresa A.; Chaky, Julian; Deshmukh, Rupesh; Vuong, Tri D.; Song, Li; Cregan, Perry B.; Nelson, James C.; Shannon, J. Grover; Specht, James E.; Nguyen, Henry T.
Cultivated soybean (Glycine max L.) cv. Dunbar (PI 552538) and wild G. soja (PI 326582A) exhibited significant differences in root architecture and root-related traits. In this study, phenotypic variability for root traits among 251 BC2F5 backcross inbred lines (BILs) developed from the cross Dunbar/PI 326582A were identified. The root systems of the parents and BILs were evaluated in controlled environmental conditions using a cone system at seedling stage. The G. max parent Dunbar contributed phenotypically favorable alleles at a major quantitative trait locus on chromosome 8 (Satt315-I locus) that governed root traits (tap root length and lateral root number) and shoot length. This QTL accounted for >10% of the phenotypic variation of both tap root and shoot length. This QTL region was found to control various shoot- and root-related traits across soybean genetic backgrounds. Within the confidence interval of this region, eleven transcription factors (TFs) were identified. Based on RNA sequencing and Affymetrix expression data, key TFs including MYB, AP2-EREBP and bZIP TFs were identified in this QTL interval with high expression in roots and nodules. The backcross inbred lines with different parental allelic combination showed different expression pattern for six transcription factors selected based on their expression pattern in root tissues. It appears that the marker interval Satt315–I locus on chromosome 8 contain an essential QTL contributing to early root and shoot growth in soybean. PMID:25756528
Van Acker, Rene; Rajcan, Istvan; Swanton, Clarence J.
Identity recognition systems allow plants to tailor competitive phenotypes in response to the genetic relatedness of neighbours. There is limited evidence for the existence of recognition systems in crop species and whether they operate at a level that would allow for identification of different degrees of relatedness. Here, we test the responses of commercial soya bean cultivars to neighbours of varying genetic relatedness consisting of other commercial cultivars (intraspecific), its wild progenitor Glycine soja, and another leguminous species Phaseolus vulgaris (interspecific). We found, for the first time to our knowledge, that a commercial soya bean cultivar, OAC Wallace, showed identity recognition responses to neighbours at different levels of genetic relatedness. OAC Wallace showed no response when grown with other commercial soya bean cultivars (intra-specific neighbours), showed increased allocation to leaves compared with stems with wild soya beans (highly related wild progenitor species), and increased allocation to leaves compared with stems and roots with white beans (interspecific neighbours). Wild soya bean also responded to identity recognition but these responses involved changes in biomass allocation towards stems instead of leaves suggesting that identity recognition responses are species-specific and consistent with the ecology of the species. In conclusion, elucidating identity recognition in crops may provide further knowledge into mechanisms of crop competition and the relationship between crop density and yield. PMID:28280587
Win, Joe; Morgan, William; Bos, Jorunn; Krasileva, Ksenia V; Cano, Liliana M; Chaparro-Garcia, Angela; Ammar, Randa; Staskawicz, Brian J; Kamoun, Sophien
Oomycete plant pathogens deliver effector proteins inside host cells to modulate plant defense circuitry and to enable parasitic colonization. These effectors are defined by a conserved motif, termed RXLR (for Arg, any amino acid, Leu, Arg), that is located downstream of the signal peptide and that has been implicated in host translocation. Because the phenotypes of RXLR effectors extend to plant cells, their genes are expected to be the direct target of the evolutionary forces that drive the antagonistic interplay between pathogen and host. We used the draft genome sequences of three oomycete plant pathogens, Phytophthora sojae, Phytophthora ramorum, and Hyaloperonospora parasitica, to generate genome-wide catalogs of RXLR effector genes and determine the extent to which these genes are under positive selection. These analyses revealed that the RXLR sequence is overrepresented and positionally constrained in the secretome of Phytophthora relative to other eukaryotes. The three examined plant pathogenic oomycetes carry complex and diverse sets of RXLR effector genes that have undergone relatively rapid birth and death evolution. We obtained robust evidence of positive selection in more than two-thirds of the examined paralog families of RXLR effectors. Positive selection has acted for the most part on the C-terminal region, consistent with the view that RXLR effectors are modular, with the N terminus involved in secretion and host translocation and the C-terminal domain dedicated to modulating host defenses inside plant cells.
Freire, Luísa; Passamani, Fabiana Reinis Franca; Thomas, Ariela Betsy; Nassur, Rita de Cássia Mirela Resende; Silva, Lais Mesquita; Paschoal, Fabiano Narciso; Pereira, Giuliano Elias; Prado, Guilherme; Batista, Luís Roberto
The incidence of filamentous fungi and toxin levels in grapes and wines varies depending on the variety of grapes, the wine region, agricultural practices, weather conditions, the harvest and the winemaking process. In this sense, the objective of this study was to evaluate the diversity of Aspergillus and Penicillium fungi isolated from wine grapes of the semi-arid tropical region of Brazil, evaluate the presence of ochratoxin A (OTA) in the experimental wine and verify if there is a correlation between occurrence of these fungi and the physicochemical characteristics of the wine grapes grown in the region. For the isolation of fungi we used the direct plating technique. The presence of OTA in the experimental wine was detected by high-performance liquid chromatography. The species found were Aspergillus niger, A. carbonarius, A. aculeatus, A. niger Aggregate, A. flavus, A. sojae, Penicillium sclerotiorum, P. citrinum, P. glabrum, P. decumbens, P. solitum and P. implicatum. All isolates of A. carbonarius were OTA producers and all P. citrinum were citrinin producers. The highest concentration of OTA was found in red wine (0.29μg/L). All species identified in this study, except A. flavus, showed a positive correlation with at least one physicochemical parameter assessed, highlighting the pectin content, total sugar, total acidity and phenolic compounds.
Tyler, Brett M; Kale, Shiv D; Wang, Qunqing; Tao, Kai; Clark, Helen R; Drews, Kelly; Antignani, Vincenzo; Rumore, Amanda; Hayes, Tristan; Plett, Jonathan M; Fudal, Isabelle; Gu, Biao; Chen, Qinghe; Affeldt, Katharyn J; Berthier, Erwin; Fischer, Gregory J; Dou, Daolong; Shan, Weixing; Keller, Nancy P; Martin, Francis; Rouxel, Thierry; Lawrence, Christopher B
A wide diversity of pathogens and mutualists of plant and animal hosts, including oomycetes and fungi, produce effector proteins that enter the cytoplasm of host cells. A major question has been whether or not entry by these effectors can occur independently of the microbe or requires machinery provided by the microbe. Numerous publications have documented that oomycete RxLR effectors and fungal RxLR-like effectors can enter plant and animal cells independent of the microbe. A recent reexamination of whether the RxLR domain of oomycete RxLR effectors is sufficient for microbe-independent entry into host cells concluded that the RxLR domains of Phytophthora infestans Avr3a and of P. sojae Avr1b alone are NOT sufficient to enable microbe-independent entry of proteins into host and nonhost plant and animal cells. Here, we present new, more detailed data that unambiguously demonstrate that the RxLR domain of Avr1b does show efficient and specific entry into soybean root cells and also into wheat leaf cells, at levels well above background nonspecific entry. We also summarize host cell entry experiments with a wide diversity of oomycete and fungal effectors with RxLR or RxLR-like motifs that have been independently carried out by the seven different labs that coauthored this letter. Finally we discuss possible technical reasons why specific cell entry may have been not detected by Wawra et al. (2013).
Piotrowski, Jeff S.; Okada, Hiroki; Lu, Fachuang; Li, Sheena C.; Hinchman, Li; Ranjan, Ashish; Smith, Damon L.; Higbee, Alan J.; Ulbrich, Arne; Coon, Joshua J.; Deshpande, Raamesh; Bukhman, Yury V.; McIlwain, Sean; Ong, Irene M.; Myers, Chad L.; Boone, Charles; Landick, Robert; Ralph, John; Kabbage, Mehdi; Ohya, Yoshikazu
A rise in resistance to current antifungals necessitates strategies to identify alternative sources of effective fungicides. We report the discovery of poacic acid, a potent antifungal compound found in lignocellulosic hydrolysates of grasses. Chemical genomics using Saccharomyces cerevisiae showed that loss of cell wall synthesis and maintenance genes conferred increased sensitivity to poacic acid. Morphological analysis revealed that cells treated with poacic acid behaved similarly to cells treated with other cell wall-targeting drugs and mutants with deletions in genes involved in processes related to cell wall biogenesis. Poacic acid causes rapid cell lysis and is synergistic with caspofungin and fluconazole. The cellular target was identified; poacic acid localized to the cell wall and inhibited β-1,3-glucan synthesis in vivo and in vitro, apparently by directly binding β-1,3-glucan. Through its activity on the glucan layer, poacic acid inhibits growth of the fungi Sclerotinia sclerotiorum and Alternaria solani as well as the oomycete Phytophthora sojae. A single application of poacic acid to leaves infected with the broad-range fungal pathogen S. sclerotiorum substantially reduced lesion development. In conclusion, the discovery of poacic acid as a natural antifungal agent targeting β-1,3-glucan highlights the potential side use of products generated in the processing of renewable biomass toward biofuels as a source of valuable bioactive compounds and further clarifies the nature and mechanism of fermentation inhibitors found in lignocellulosic hydrolysates.
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
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.
Han, Yingpeng; Zhao, Xue; Liu, Dongyuan; Li, Yinghui; Lightfoot, David A; Yang, Zhijiang; Zhao, Lin; Zhou, Gang; Wang, Zhikun; Huang, Long; Zhang, Zhiwu; Qiu, Lijuan; Zheng, Hongkun; Li, Wenbin
Present-day soybeans consist of elite cultivars and landraces (Glycine max, fully domesticated (FD)), annual wild type (Glycine soja, nondomesticated (ND)), and semi-wild type (semi-domesticated (SD)). FD soybean originated in China, although the details of its domestication history remain obscure. More than 500 diverse soybean accessions were sequenced using specific-locus amplified fragment sequencing (SLAF-seq) to address fundamental questions regarding soybean domestication. In total, 64,141 single nucleotide polymorphisms (SNPs) with minor allele frequencies (MAFs) > 0.05 were found among the 512 tested accessions. The results indicated that the SD group is not a hybrid between the FD and ND groups. The initial domestication region was pinpointed to central China (demarcated by the Great Wall to the north and the Qinling Mountains to the south). A total of 800 highly differentiated genetic regions and > 140 selective sweeps were identified, and these were three- and twofold more likely, respectively, to encompass a known quantitative trait locus (QTL) than the rest of the soybean genome. Forty-three potential quantitative trait nucleotides (QTNs; including 15 distinct traits) were identified by genome-wide association mapping. The results of the present study should be beneficial for soybean improvement and provide insight into the genetic architecture of traits of agronomic importance.
Laten, Howard M; Havecker, Ericka R; Farmer, Lisa M; Voytas, Daniel F
SIRE1 is unusual among Ty1-copia retrotransposons in that it has an additional open reading frame with structural features similar to retroviral envelope proteins between pol and the 3' LTR. Here we report the characterization and comparison of eight different SIRE1 elements derived from a soybean genomic library, as well as SIRE1 reverse transcriptases from Glycine soja. The DNA sequences of the eight SIRE1 elements are highly homogeneous and share greater than 95% nucleotide identity. Partial sequences obtained from BAC ends are similarly conserved. Phylogenetic analyses resolve two closely related SIRE1 lineages, and nucleotide changes within and between SIRE1 lineages have occurred to preserve function. Both the gag and the env-like genes are evolving under similar levels of functional constraint. Considerable sequence heterogeneity in the form of short duplications was found within the LTRs and in the region between the envelope-like ORF and the 3' LTR. These duplications are suggestive of slippage by reverse transcriptase during replication. Sequence identity between LTRs of individual insertions suggests that they transposed within the past 70,000 years. Two of 10 SIRE1 insertions examined abut Ty3-gypsy retroelements. Since the soybean genome harbors more than 1,000 SIRE1 insertions, the collective data suggest that SIRE1 has undergone a very recent and robust amplification in soybean.
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
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
Sterken, V. J.; Strub, P.; Soja, R. H.; Srama, R.; Krüger, H.; Grün, E.
Beyond 3 AU heliocentric distance, the flow of interstellar dust through the solar system is a dominant component of the total dust population. The modulation of this flux with the solar cycle and the position in the solar system has been predicted by theoretical studies since the seventies. The modulation was proven to exist by matching dust trajectory simulations with real spacecraft data from Ulysses in 1998. The modulations were further analyzed and studies in detail in 2012. The current ESA interplanetary meteoroid model IMEM includes an interstellar dust component, but this component was modelled only with straight line trajectories through the solar system. For the new ESA IMEX model, a high-resolution interstellar dust component is implemented separately from a dust streams module. The dust streams module focuses on dust in streams that was released from comets (cf. Abstract R. Soja). Parallel processing techniques are used to improve computation time (cf. Abstract P. Strub). The goal is to make predictions for the interstellar dust flux as close to the Sun as 1 AU or closer, for future space mission design.
Tsubokura, Yasutaka; Hajika, Makita; Kanamori, Hiroyuki; Xia, Zhengjun; Watanabe, Satoshi; Kaga, Akito; Katayose, Yuichi; Ishimoto, Masao; Harada, Kyuya
β-conglycinin, a major seed protein in soybean, is composed of α, α', and β subunits sharing a high homology among them. Despite its many health benefits, β-conglycinin has a lower amino acid score and lower functional gelling properties compared to glycinin, another major soybean seed protein. In addition, the α, α', and β subunits also contain major allergens. A wild soybean (Glycine soja Sieb et Zucc.) line, 'QT2', lacks all of the β-conglycinin subunits, and the deficiency is controlled by a single dominant gene, Scg-1 (Suppressor of β-conglycinin). This gene was characterized using a soybean cultivar 'Fukuyutaka', 'QY7-25', (its near-isogenic line carrying the Scg-1 gene), and the F₂ population derived from them. The physical map of the Scg-1 region covered by lambda phage genomic clones revealed that the two α-subunit genes, a β-subunit gene, and a pseudo α-subunit gene were closely organized. The two α-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 α subunit. These results strongly suggest that β-conglycinin deficiency in QT2 is controlled by post-transcriptional gene silencing through the inverted repeat of the α subunits.
Hua, Chenlei; Meijer, Harold J G; de Keijzer, Jeroen; Zhao, Wei; Wang, Yuanchao; Govers, Francine
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.
Li, Qing-Tian; Lu, Xiang; Song, Qingxin; Chen, Hao-Wei; Wei, Wei; Tao, Jian-Jun; Bian, Xiao-Hua; Shen, Ming; Ma, Biao; Zhang, Wan-Ke; Bi, Ying-Dong; Li, Wei; Lai, Yong-Cai; Lam, Sin Man; Shui, Guanghou; Chen, Shou-Yi; Zhang, Jin-Song
Seed oil is a momentous agronomical trait of soybean targeted by domestication in breeding. Although multiple oil-related genes have been uncovered, the knowledge of regulatory mechanism of seed oil biosynthesis is currently limited. We demonstrate that seed-preferred gene GmZF351 encoding tandem CCCH zinc finger protein is selected during domestication. Further analysis shows that GmZF351 facilitates oil accumulation by directly activating WRI1, BCCP2, KASIII, TAG1 and OLEO2 in transgenic Arabidopsis seeds. Overexpression of GmZF351 in transgenic soybean also activates lipid biosynthesis genes, thereby accelerating seed oil accumulation. ZF351 haplotype from Glycine max group and Glycine soja subgroup III correlates well with high gene expression level, seed oil contents and promoter activity, suggesting that selection of GmZF351 expression leads to increased seed oil content in cultivated soybean. Our study provides novel insights into the regulatory mechanism for seed oil accumulation and manipulation of GmZF351 may have great potential in improvement of oil production in soybean and other related crops.
Dissanayake, Asha J; Liu, Mei; Zhang, Wei; Chen, Zhen; Udayanga, Dhanushka; Chukeatirote, Ekachai; Li, XingHong; Yan, JiYe; Hyde, Kevin D
Trunk diseases in grapevine (Vitis spp.) are major problems in the wine and table-grape industries reducing the productivity, quality and longevity of vineyards. Species of Diaporthe are important fungal pathogens of grapevine trunk disease worldwide. A survey of 14 grape vineyards located in different provinces of China was yielded Diaporthe isolates associated with symptomatic grapevine wood. These isolates were identified based on morphology and a combined data matrix of rDNA ITS, partial sequences of translation elongation factor 1-α (EF 1-α), β-tubulin (TUB) and calmodulin (CAL) gene regions. Four species of Diaporthe were identified, which included Diaporthe eres, Diaporthe hongkongensis, Diaporthe phaseolorum and Diaporthe sojae. All isolates of Diaporthe caused disease on detached grape shoots in pathogenicity experiments but differed in virulence. The incidence in local vineyards and the pathogenicity results indicate that D. eres is an important pathogen of grapevine in Chinese vineyards, where it may significantly limit grape production. This is the first detailed report of Diaporthe species associated with grapevine trunk diseases in China with morphology, pathogenicity and molecular data.
Govindarajulu, Manjula; Kim, Sung-Yong; Libault, Marc; Berg, R. Howard; Tanaka, Kiwamu; Stacey, Gary; Taylor, Christopher G.
Apyrases are non-energy-coupled nucleotide phosphohydrolases that hydrolyze nucleoside triphosphates and nucleoside diphosphates to nucleoside monophosphates and orthophosphates. GS52, a soybean (Glycine soja) ecto-apyrase, was previously shown to be induced very early in response to inoculation with the symbiotic bacterium Bradyrhizobium japonicum. Overexpression of the GS52 ecto-apyrase in Lotus japonicus increased the level of rhizobial infection and enhanced nodulation. These data suggest a critical role for the GS52 ecto-apyrase during nodulation. To further investigate the role of GS52 during nodulation, we used RNA interference to silence GS52 expression in soybean (Glycine max) roots using Agrobacterium rhizogenes-mediated root transformation. Transcript levels of GS52 were significantly reduced in GS52 silenced roots and these roots exhibited reduced numbers of mature nodules. Development of the nodule primordium and subsequent nodule maturation was significantly suppressed in GS52 silenced roots. Transmission electron micrographs of GS52 silenced root nodules showed that early senescence and infected cortical cells were devoid of symbiosome-containing bacteroids. Application of exogenous adenosine diphosphate to silenced GS52 roots restored nodule development. Restored nodules contained bacteroids, thus indicating that extracellular adenosine diphosphate is important during nodulation. These results clearly suggest that GS52 ecto-apyrase catalytic activity is critical for the early B. japonicum infection process, initiation of nodule primordium development, and subsequent nodule organogenesis in soybean. PMID:19036836
Cerovský, V; Tichá, M; Horejsi, V; Kocourek, J
p-Aminophenyl glycosides and glycosylamines were coupled to periodate oxidized Dextran T-500 either directly or through an epsilon-aminocaproic acid spacer. The new glycosylated derivatives of dextran specifically precipitate lectins having the appropriate carbohydrate specificity, and thus were used in the preparation of affinity gels for affinity electrophoresis of lectins. The apparent strength of interaction of several lectins with carbohydrate residues immobilized in this way was less than with carbohydrates immobilized in O-glycosyl polyacrylamide copolymers. The presence of epsilon-aminocaproic spacer had no effect on the strength of interaction. The advantages of this type of macromolecular derivative of the ligand for affinity electrophoresis and some differences between the glycosylated dextrans and O-glycosyl polyacrylamide copolymers are discussed. Dextrans containing bound p-aminophenyl alpha-D-mannopyranoside and p-aminophenyl alpha-D-glucopyranoside were used to study the binding properties of concanavalin A and the lectin from Lathyrus sativus seeds. For the investigation of interaction of lectins from Ricinus communis and Glycine soja seeds, dextran derivatives containing bound p-aminophenyl alpha- and beta-D-galactopyranosides and alpha- and beta-D-galactopyranosylamines were used.
Sakamoto, Tatsuji; Ogura, Atsuhiro; Inui, Misako; Tokuda, Sayaka; Hosokawa, Sachiko; Ihara, Hideshi; Kasai, Naoya
An arabinoxylan arabinofuranohydrolase (AXS5) was purified from the culture filtrate of Penicillium chrysogenum 31B. A cDNA encoding AXS5 (axs5) was isolated by in vitro cloning using the N-terminal amino acid sequence of the native enzyme as a starting point. The deduced amino acid sequence of the axs5 gene has high similarities with those of arabinoxylan arabinofuranohydrolases of Aspergillus niger, Aspergillus tubingensis, and Aspergillus sojae. Module sequence analysis revealed that a "Glyco_hydro_62" was present at position 28-299 of AXS5. This is a family of α-L-arabinofuranosidases which are all members of glycoside hydrolase family 62. Recombinant AXS5 (rAXS5) expressed in Escherichia coli was highly active on arabinoxylan but not on branched sugar beet arabinan. (1)H-NMR analysis revealed that the rAXS5 cleaved arabinosyl side-chains linked to C-2 and C-3 of single-substituted xylose residues in arabinoxylan. Semi-quantitative RT-PCR analysis indicated that expression of the axs5 gene in P. chrysogenum 31B was strongly induced by adding D-xylose and arabinoxylan to the culture medium. Moreover, two binding sites of XlnR, a transcriptional activator that regulates the expression of the genes encoding xylanolytic enzymes, are present in the upstream region of the axs5 gene. These results suggest that AXS5 is involved in xylan degradation.
Yan, Qiang; Cui, Xiaoxia; Lin, Shuai; Gan, Shuping; Xing, Han; Dou, Daolong
The cytochrome P450 monooxygenases (P450s) represent a large and important enzyme superfamily in plants. They catalyze numerous monooxygenation/hydroxylation reactions in biochemical pathways, P450s are involved in a variety of metabolic pathways and participate in the homeostasis of phytohormones. The CYP82 family genes specifically reside in dicots and are usually induced by distinct environmental stresses. However, their functions are largely unknown, especially in soybean (Glycine max L.). Here, we report the function of GmCYP82A3, a gene from soybean CYP82 family. Its expression was induced by Phytophthora sojae infection, salinity and drought stresses, and treatment with methyl jasmonate (MeJA) or ethephon (ETH). Its expression levels were consistently high in resistant cultivars. Transgenic Nicotiana benthamiana plants overexpressing GmCYP82A3 exhibited strong resistance to Botrytis cinerea and Phytophthora parasitica, and enhanced tolerance to salinity and drought stresses. Furthermore, transgenic plants were less sensitive to jasmonic acid (JA), and the enhanced resistance was accompanied with increased expression of the JA/ET signaling pathway-related genes. PMID:27588421
Liu, Chun; Cheng, Fenfen; Sun, Yingen; Ma, Hongyu; Yang, Xiaoquan
An alkaline isoform of the PR-5 protein (designated GmOLPc) has been purified from soybean hulls and identified by MALDI-TOF/TOF-MS. GmOLPc effectively inhibited in vitro the growth of Phytophthora soja spore and Pseudomonas syringae pv glycinea. The antimicrobial activity of GmOLPc should be mainly ascribed to its high binding affinity with vesicles composed of DPPG, (1,3)-β-D-glucans, and weak endo-(1,3)-β-D-glucanase activity. From the 3D models, predicted by the homology modeling, GmOLPc contains an extended negatively charged cleft. The cleft was proved to be a prerequisite for endo-(1,3)-β-D-glucanase activity. Molecular docking revealed that the positioning of linear (1,3)-β-D-glucans in the cleft of GmOLPc allowed an interaction with Glu83 and Asp101 that were responsible for the hydrolytic cleavage of glucans. Interactions of GmOLPc with model membranes indicated that GmOLPc possesses good surface activity which could contribute to its antimicrobial activity, as proved by the behavior of perturbing the integrity of membranes through surface hydrophobic amino acid residues (Phe89 and Phe94).
Overdijk, Elysa J R; DE Keijzer, Jeroen; DE Groot, Deborah; Schoina, Charikleia; Bouwmeester, Klaas; Ketelaar, Tijs; Govers, Francine
Live-cell imaging of plant-pathogen interactions is often hampered by the tissue complexity and multicell layered nature of the host. Here, we established a novel pathosystem with the moss Physcomitrella patens as host for Phytophthora. The tip-growing protonema cells of this moss are ideal for visualizing interactions with the pathogen over time using high-resolution microscopy. We tested four Phytophthora species for their ability to infect P. patens and showed that P. sojae and P. palmivora were only rarely capable to infect P. patens. In contrast, P. infestans and P. capsici frequently and successfully penetrated moss protonemal cells, showed intracellular hyphal growth and formed sporangia. Next to these successful invasions, many penetration attempts failed. Here the pathogen was blocked by a barrier of cell wall material deposited in papilla-like structures, a defence response that is common in higher plants. Another common response is the upregulation of defence-related genes upon infection and also in moss we observed this upregulation in tissues infected with Phytophthora. For more advanced analyses of the novel pathosystem we developed a special set-up that allowed live-cell imaging of subcellular defence processes by high-resolution microscopy. With this set-up, we revealed that Phytophthora infection of moss induces repositioning of the nucleus, accumulation of cytoplasm and rearrangement of the actin cytoskeleton, but not of microtubules.
Zhang, Huajian; Wu, Qun; Cao, Shun; Zhao, Tongyao; Chen, Ling; Zhuang, Peitong; Zhou, Xiuhong; Gao, Zhimou
In this study, we report the cloning of the SsCut gene encoding cutinase from Sclerotinia sclerotiorum. We isolated a 609-bp cDNA encoding a polypeptide of 202 amino acids with a molecular weight of 20.4 kDa. Heterologous expression of SsCut in Escherichia coli (His-SsCut) caused the formation of lesions in tobacco that closely resembled hypersensitive response lesions. Mutational analysis identified the C-terminal-half peptide and the same amino acids indispensable for both enzyme and elicitor activity. His-SsCut was caused cell death in Arabidopsis, soybean (Glycine max), oilseed rape (Brassica napus), rice (Oryza sativa), maize (Zea mays), and wheat (Triticum aestivum), indicating that both dicot and monocot species are responsive to the elicitor. Furthermore, the elicitation of tobacco was effective in the induction of the activities of hydrogen peroxide, phenylalanine ammonia-lyase, peroxides, and polyphenol oxidase. His-SsCut-treated plants exhibited enhanced resistance as indicated by a significant reduction in the number and size of S. sclerotiorum, Phytophthora sojae, and P. nicotianae lesions on leaves relative to controls. Real-time PCR results indicated that the expression of defense-related genes and genes involved in signal transduction were induced by His-SsCut. Our results demonstrate that SsCut is an elicitor that triggers defense responses in plants and will help to clarify its relationship to downstream signaling pathways that induce defense responses.
Li, Qing-Tian; Lu, Xiang; Song, Qing-Xin; Chen, Hao-Wei; Wei, Wei; Tao, Jian-Jun; Ma, Biao; Bi, Ying-Dong; Li, Wei; Lai, Yong-Cai; Shui, Guang-Hou; Chen, Shou-Yi
Seed oil is a momentous agronomical trait of soybean (Glycine max) targeted by domestication in breeding. Although multiple oil-related genes have been uncovered, knowledge of the regulatory mechanism of seed oil biosynthesis is currently limited. We demonstrate that the seed-preferred gene GmZF351, encoding a tandem CCCH zinc finger protein, is selected during domestication. Further analysis shows that GmZF351 facilitates oil accumulation by directly activating WRINKLED1, BIOTIN CARBOXYL CARRIER PROTEIN2, 3-KETOACYL-ACYL CARRIER PROTEIN SYNTHASE III, DIACYLGLYCEROL O-ACYLTRANSFERASE1, and OLEOSIN2 in transgenic Arabidopsis (Arabidopsis thaliana) seeds. Overexpression of GmZF351 in transgenic soybean also activates lipid biosynthesis genes, thereby accelerating seed oil accumulation. The ZF351 haplotype from the cultivated soybean group and the wild soybean (Glycine soja) subgroup III correlates well with high gene expression level, seed oil contents and promoter activity, suggesting that selection of GmZF351 expression leads to increased seed oil content in cultivated soybean. Our study provides novel insights into the regulatory mechanism for seed oil accumulation, and the manipulation of GmZF351 may have great potential in the improvement of oil production in soybean and other related crops. PMID:28184009
Molinari, Chiara; Lehsten, Veiko; Blarquez, Olivier; Clear, Jennifer; Carcaillet, Christopher; Bradshaw, Richard HW
Forest fire is one of the most critical ecosystem processes in the boreal megabiome, and it is likely that its frequency, size and severity have had a primary role in vegetation dynamics since the Last Ice Age (Kasischke & Stocks 2000). Fire not only organizes the physical and biological attributes of boreal forests, but also affects biogeochemical cycling, particularly the carbon balance (Balshi et al. 2007). Due to their location at climatically sensitive northern latitudes, boreal forests are likely to be significantly affected by global warming with a consequent increase in biomass burning (Soja et al. 2007), a variation in vegetation structure and composition (Johnstone et al. 2004) and a rise in atmospheric carbon dioxide concentration (Bond-Lamberty et al. 2007). Even if the ecological role of wildfire in boreal forest is widely recognized, a clearer understanding of the environmental factors controlling fire dynamics and how variations in fire regimes impact forest ecosystems is essential in order to place modern fire processes in a meaningful context for projecting ecosystem behaviour in a changing environment (Kelly et al. 2013). Because fire return intervals and successional cycles in boreal forests occur over decadal to centennial timescales (Hu et al. 2006), palaeoecological research seems to be one of the most promising tool for elucidating ecosystem changes over a broad range of environmental conditions and temporal scales. Within this context, our first aim is to reconstruct spatial and temporal patterns of boreal forests fire dynamics during the Holocene based on sedimentary charcoal records. As a second step, trends in biomass burning will be statistically analysed in order to disentangle between regional and local drivers. The use of European and north-American sites will give us the unique possibility to perform a large scale analysis on one of the broadest biome in the world and to underline the different patterns of fire in these two
Nautiyal, C. S.; Hegde, S. V.; van Berkum, P.
The pigeon pea strains of Bradyrhizobium CC-1, CC-8, UASGR(S), and F4 were evaluated for nodulation, effectiveness for N2 fixation, and H2 oxidation with homologous and nonhomologous host plants. Strain CC-1 nodulated Macroptilium atropurpureum, Vigna unguiculata, Glycine max, and G. soja but did not nodulate Pisum sativum, Phaseolus vulgaris, Trigonella foenum-graecum, and Trifolium repens. Strain F4 nodulated G. max cv. Peking and PI 434937 (Malayan), but the symbioses formed were poor. Similarly, G. max cv. Peking, cv. Bragg, PI 434937, PR 13-28-2-8-7, and HM-1 were nodulated by strain CC-1, and symbioses were also poor. G. max cv. Williams and cv. Clark were not nodulated. H2 uptake activity was expressed with pigeon pea and cowpea, but not with soybean. G. max cv. Bragg grown in Bangalore, India, in local soil not previously exposed to Bradyrhizobium japonicum formed nodules with indigenous Bradyrhizobium spp. Six randomly chosen isolates, each originating from a different nodule, formed effective symbioses with pigeon pea host ICPL-407, nodulated PR 13-28-2-8-7 soybean forming moderately effective symbioses, and did not nodulate Williams soybean. These results indicate the six isolates to be pigeon pea strains although they originated from soybean nodules. Host-determined nodulation of soybean by pigeon pea Bradyrhizobium spp. may depend upon the ancestral backgrounds of the cultivars. The poor symbioses formed by the pigeon pea strains with soybean indicate that this crop should be inoculated with B. japonicum for its cultivation in soils containing only pigeon pea Bradyrhizobium spp. PMID:16347542
Krishnaraj, Chandran; Harper, Stacey L; Choe, Ho Sung; Kim, Kwang-Pyo; Yun, Soon-Il
In the present study, silver nanoparticles (AgNPs) synthesized from aqueous leaves extract of Malva crispa and their mode of interaction with food- and water-borne microbes were investigated. Formation of AgNPs was conformed through UV-Vis, FE-SEM, EDS, AFM, and HR-TEM analyses. Further the concentration of silver (Ag) in the reaction mixture was conformed through ICP-MS analysis. Different concentration of nanoparticles (1-3 mM) tested to know the inhibitory effect of bacterial pathogens such as Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Salmonella typhi, Salmonella enterica and the fungal pathogens of Penicillium expansum, Penicillium citrinum, Aspergillus oryzae, Aspergillus sojae and Aspergillus niger. Interestingly, nanoparticles synthesized from 2 to 3 mM concentration of AgNO3 showed excellent inhibitory activities against both bacterial and fungal pathogens which are well demonstrated through well diffusion, poison food technique, minimum inhibitory concentration (MIC), and minimum fungicidal concentration (MFC). In addition, mode of interaction of nanoparticles into both bacterial and fungal pathogens was documented through Bio-TEM analysis. Further the genomic DNA isolated from test bacterial strains and their interaction with nanoparticles was carried out to elucidate the possible mode of action of nanoparticles against bacteria. Interestingly, AgNPs did not show any genotoxic effect against all the tested bacterial strains which are pronounced well in agarose gel electrophoresis and for supporting this study, UV-Vis and Bio-TEM analyses were carried out in which no significant changes observed compared with control. Hence, the overall results concluded that the antimicrobial activity of biogenic AgNPs occurred without any DNA damage.
Kato, Shinya; Aoshima, Hisae; Saitoh, Yasukazu; Miwa, Nobuhiko
We previously reported biological safety of fullerene-C60 (C60) incorporated in liposome consisting of hydrogenated lecithin and glycine soja sterol, as Liposome-Fullerene (0.5% aqueous phase; a particle size, 76nm; Lpsm-Flln), and its cytoprotective activity against UVA. In the present study, Lpsm-Flln was administered on the surface of three-dimensional human skin tissue model, rinsed out before each UVA-irradiation at 4 J/cm(2), and thereafter added again, followed by 19-cycle-repetition for 4 days (sum: 76 J/cm(2)). UVA-caused corneum scaling and disruption of epidermis layer were detected by scanning electron microscopy. Breakdown of collagen type I/IV, DNA strand cleavage and pycnosis/karyorrhexis were observed in vertical cross-sections of UVA-irradiated skin models visualized with fluorescent immunostain or Hoechst 33342 stain. These skin damages were scarcely repressed by liposome alone, but appreciably repressed by Lpsm-Flln of 250 ppm, containing 0.75 ppm of C60-equivalent to a 1/3300-weight amount vs. the whole liposome. Upon administration with Lpsm-Flln [16.7 microM (12 ppm): C60-equivalent] on human abdomen skin biopsies mounted in Franz diffusion cells, C60 permeated after 24h into the epidermis at 1.86 nmol/g tissue (1.34 ppm), corresponding to 0.3% of the applied amount and a 9.0-fold dilution rate, but C60 was not detected in the dermis by HPLC, suggesting no necessity for considering a toxicity of C60 due to systemic circulation via dermal veins. Thus Lpsm-Flln has a potential to be safely utilized as a cosmetic anti-oxidative ingredient for UVA-protection.
Jang, Seo-Jung; Kim, Yu-Jin; Lee, Sul-Hee; Park, Young-Seo; Park, Jung-Min; Bai, Dong-Hoon
A Gram-stain-positive, polar flagella-containing, rod-shaped, obligate aerobic, endospore-forming bacterium, strain TK1655(T), was isolated from the traditional Korean food gochujang. The 16S rRNA sequence of strain TK1655(T) was a member of the genus Oceanobacillus similar to that of the type strain of Oceanobacillus oncorhynchi subsp. incaldanensis DSM 16557(T) (97.2%), O. oncorhynchi subsp. oncorhynchi JCM 12661(T) (97.1%), O. locisalsi KCTC 13253(T) (97.0%), and O. sojae JCM 15792(T) (96.9%). Strain TK1655(T) was oxidase and catalase positive. Colonies were circular, smooth, low convex, cream in colour, and measured about 0.5-1.0 mm in diameter. The range for growth was 20-40°C (optimal, 30°C), pH 6.0-10.0 (optimal, 7.0), and 2-16% (w/v) NaCl (optimal, 2%). Additionally, the cells contained meso-DAP, and the predominant isoprenoid quinone was MK-7. The complex polar lipids were consisted of diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylcholine (PC). The major cellular fatty acid components were iso-C15:0, anteiso-C15:0, iso-C16:0, and anteiso-C17:0, and the DNA G+C content was 40.5%. DNA-DNA relatedness of our novel strain and reference strain O. locisalsi KCTC 13253(T), O. oncorhynchi subsp. incaldanensis DSM 16557(T), O. oncorhynchi subsp. oncorhynchi JCM 12661(T) was 45.7, 43.8, and 41.9%. From the results of phenotypic, chemotaxonomic, and phylogenetic analyses of strain TK1655(T), we propose the novel species Oceanobacillus gochujangensis sp. nov. The type strain is TK1655(T) (=KCCM 101304(T) =KCTC 33014(T) =CIP 110582(T) =NBRC 109637(T)).
Bartley, G E; Viitanen, P V; Pecker, I; Chamovitz, D; Hirschberg, J; Scolnik, P A
Carotenoids are orange, yellow, or red photo-protective pigments present in all plastids. The first carotenoid of the pathway is phytoene, a colorless compound that is converted into colored carotenoids through a series of desaturation reactions. Genes coding for carotenoid desaturases have been cloned from microbes but not from plants. We report the cloning of a cDNA for pds1, a soybean (Glycine max) gene that, based on a complementation assay using the photosynthetic bacterium Rhodobacter capsulatus, codes for an enzyme that catalyzes the two desaturation reactions that convert phytoene into zeta-carotene, a yellow carotenoid. The 2281-base-pair cDNA clone analyzed contains an open reading frame with the capacity to code for a 572-residue protein of predicted Mr 63,851. Alignment of the deduced Pds1 peptide sequence with the sequences of fungal and bacterial carotenoid desaturases revealed conservation of several amino acid residues, including a dinucleotide-binding motif that could mediate binding to FAD. The Pds1 protein is synthesized in vitro as a precursor that, upon import into isolated chloroplasts, is processed to a smaller mature form. Hybridization of the pds1 cDNA to genomic blots indicated that this gene is a member of a low-copy-number gene family. One of these loci was genetically mapped using restriction fragment length polymorphisms between Glycine max and Glycine soja. We conclude that pds1 is a nuclear gene encoding a phytoene desaturase enzyme that, as its microbial counterparts, contains sequence motifs characteristic of flavoproteins. Images PMID:1862081
Hernando Requejo, Ovidio; Rubio Rodríguez, Ma Carmen
Introducción: hoy día se conoce cómo la dieta juega un importante papel tanto en la producción como en la prevención del cáncer. Métodos: se ha realizado una completa revisión bibliográfica para intentar conocer mejor aquellas dietas que pueden ser protectoras contra el cáncer o aquellas con más riesgo de producir cáncer. Resultados: entre los alimentos que pueden proteger de la aparición del cáncer destacan las frutas y verduras, también la soja y sus derivados y los pescados han mostrado protección frente a algunos tipos de tumores. Como claros factores que aumentan el riesgo de cáncer caben destacar la carne roja o procesada, los lácteos y el alcohol. El papel protector de vitaminas, antioxidantes y otros micronutrientes ha de ser explorado más a fondo. La obesidad parece un factor etiológico claro, asimismo la restricción calórica podría, de forma antagónica a la obesidad, proteger contra la aparición de tumores en estudios de laboratorio, pero se requerirán estudios clínicos para validar esta hipótesis. Conclusiones: el control de la dieta y la selección adecuada de los alimentos sería un método muy útil y de bajo coste en la prevención de tumores.
Luo, Xiao; Bai, Xi; Zhu, Dan; Li, Yong; Ji, Wei; Cai, Hua; Wu, Jing; Liu, Baohui; Zhu, Yanming
Plant acclimation to environmental stress is controlled by a complex network of regulatory genes that compose distinct stress-response regulons. The C2H2-type zinc-finger proteins (ZFPs) have been implicated in different cellular processes involved in plant development and stress responses. Through microarray analysis, an alkaline (NaHCO(3))-responsive ZFP gene GsZFP1 was identified and subsequently cloned from Glyycine soja. GsZFP1 encodes a 35.14 kDa protein with one C2H2-type zinc-finger motif. The QALGGH domain, conserved in most plant C2H2-type ZFPs, is absent in the GsZFP1 protein sequence. A subcellular localization study using a GFP fusion protein indicated that GsZFP1 is localized to the nucleus. Real-time RT-PCR analysis showed that GsZFP1 was induced in the leaf by ABA (100 μM), salt (200 mM NaCl), and cold (4°C), and in the root by ABA (100 μM), cold (4°C), and drought (30% PEG 6000). Over-expression of GsZFP1 in transgenic Arabidopsis resulted in a greater tolerance to cold and drought stress, a decreased water loss rate, and an increase in proline irrespective of environmental conditions. The over-expression of GsZFP1 also increased the expression of a number of stress-response marker genes, including CBF1, CBF2, CBF3, NCED3, COR47, and RD29A in response to cold stress and RAB18, NCED3, P5CS, RD22, and RD29A in response to drought stress, especially early during stress treatments. Our studies suggest that GsZFP1 plays a crucial role in the plant response to cold and drought stress.
Zhu, Yong-heng; Li, Ke-zhong; Zhang, Heng; Han, Fei; Zhou, Ju-hua; Gao, Ting-ting
A survey was carried out to investigate soil nematode communities in the plant associations of gramineae (Arthraxon lanceolatus, AL; Imperata cylindrica, IC) and leguminosae (Glycine soja, GS) in reclaimed land of copper-mine-tailings and in the plant associations of gramineae (Digitaria chrysoblephara, DC-CK) of peripheral control in Fenghuang Mountain, Tongling City. A total of 1277 nematodes were extracted and sorted into 51 genera. The average individual density of the nematodes was 590 individuals · 100 g(-1) dry soil. In order to analyze the distribution character- istics of soil nematode communities in reclaimed land of copper-mine-tailings, Shannon community diversity index and soil food web structure indices were applied in the research. The results showed that the total number of nematode genus and the Shannon community diversity index of soil nematode in the three plant associations of AL, IC and GS were less than that in the plant associations of DC-CK. Compared with the ecological indices of soil nematode communities among the different plant associations in reclaimed land of copper-mine-tailings and peripheral natural habitat, we found that the structure of soil food web in the plant associations of GS was more mature, with bacterial decomposition being dominant in the soil organic matter decomposition, and that the soil ecosystem in the plant associations of GS was not stable with low interference. This indicated that the soil food web in the plant associations of leguminosae had a greater development potential to improve the ecological stability of the reclaimed land of copper-mine-tailings. On the other hand, the structure of soil food web in the plant associations of AL and IC were relatively stable in a structured state with fungal decomposition being dominant in the decomposition of soil organic matter. This indicated that the soil food web in the plant associations of gramineae was at a poor development level.
Abad, Z Gloria; Abad, Jorge A; Cacciola, Santa Olga; Pane, Antonella; Faedda, Roberto; Moralejo, Eduardo; Pérez-Sierra, Ana; Abad-Campos, Paloma; Alvarez-Bernaola, Luis A; Bakonyi, József; Józsa, András; Herrero, Maria Luz; Burgess, Treena I; Cunnington, James H; Smith, Ian W; Balci, Yilmaz; Blomquist, Cheryl; Henricot, Béatrice; Denton, Geoffrey; Spies, Chris; Mcleod, Adele; Belbahri, Lassaad; Cooke, David; Kageyama, Koji; Uematsu, Seiji; Kurbetli, Ilker; Değirmenci, Kemal
A non-papillate, heterothallic Phytophthora species first isolated in 2001 and subsequently from symptomatic roots, crowns and stems of 33 plant species in 25 unrelated botanical families from 13 countries is formally described here as a new species. Symptoms on various hosts included crown and stem rot, chlorosis, wilting, leaf blight, cankers and gumming. This species was isolated from Australia, Hungary, Israel, Italy, Japan, the Netherlands, Norway, South Africa, Spain, Taiwan, Turkey, the United Kingdom and United States in association with shrubs and herbaceous ornamentals grown mainly in greenhouses. The most prevalent hosts are English ivy (Hedera helix) and Cistus (Cistus salvifolius). The association of the species with acorn banksia (Banksia prionotes) plants in natural ecosystems in Australia, in affected vineyards (Vitis vinifera) in South Africa and almond (Prunus dulcis) trees in Spain and Turkey in addition to infection of shrubs and herbaceous ornamentals in a broad range of unrelated families are a sign of a wide ecological adaptation of the species and its potential threat to agricultural and natural ecosystems. The morphology of the persistent non-papillate ellipsoid sporangia, unique toruloid lobate hyphal swellings and amphigynous antheridia does not match any of the described species. Phylogenetic analysis based on sequences of the ITS rDNA, EF-1α, and β-tub supported that this organism is a hitherto unknown species. It is closely related to species in ITS clade 7b with the most closely related species being P. sojae. The name Phytophthora niederhauserii has been used in previous studies without the formal description of the holotype. This name is validated in this manuscript with the formal description of Phytophthora niederhauserii Z.G. Abad et J.A. Abad, sp. nov. The name is coined to honor Dr John S. Niederhauser, a notable plant pathologist and the 1990 World Food Prize laureate.
Sundaramoorthy, Jagadeesh; Park, Gyu Tae; Chang, Jeong Ho; Lee, Jeong-Dong; Kim, Jeong Hoe; Seo, Hak Soo; Chung, Gyuhwa; Song, Jong Tae
In soybean, flavonoid 3′5′-hydroxylase (F3′5′H) and dihydroflavonol-4-reductase (DFR) play a crucial role in the production of anthocyanin pigments. Loss-of-function of the W1 locus, which encodes the former, or W3 and W4, which encode the latter, always produces white flowers. In this study, we searched for new genetic components responsible for the production of white flowers in soybean and isolated four white-flowered mutant lines, i.e., two Glycine soja accessions (CW12700 and CW13381) and two EMS-induced mutants of Glycine max (PE1837 and PE636). F3′5′H expression in CW12700, PE1837, and PE636 was normal, whereas that in CW13381 was aberrant and missing the third exon. Sequence analysis of F3′5′H of CW13381 revealed the presence of an indel (~90-bp AT-repeat) in the second intron. In addition, the F3′5′H of CW12700, PE1837, and PE636 harbored unique single-nucleotide substitutions. The single nucleotide polymorphisms resulted in substitutions of amino acid residues located in or near the SRS4 domain of F3′5′H, which is essential for substrate recognition. 3D structure modeling of F3′5′H indicated that the substitutions could interfere with an interaction between the substrate and heme group and compromise the conformation of the active site of F3′5′H. Recombination analysis revealed a tight correlation between all of the mutant alleles at the W1 locus and white flower color. On the basis of the characterization of the new mutant alleles, we discussed the biological implications of F3′5′H and DFR in the determination of flower colors in soybean. PMID:27442124
Simpore, Jacques; Kabore, Fatoumata; Zongo, Frederic; Dansou, Deleli; Bere, Augustin; Pignatelli, Salvatore; Biondi, Daniela M; Ruberto, Giuseppe; Musumeci, Salvatore
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. PMID:16430775
Liu, Y; Dammann, C; Bhattacharyya, M K
Matrix metalloproteinases (MMPs) play an important role in host defense responses against pathogens in mammals where their activities lead to the production of antimicrobial peptides. We have identified a novel soybean (Glycine max) metalloproteinase gene, GmMMP2, that is transcriptionally up-regulated in infected tissues. The deduced amino acid sequence indicates that this gene belongs to the MMP family. It is a preproprotein containing an N-terminal signal peptide, a cysteine switch, a zinc-binding catalytic motif, and a C-terminal transmembrane domain. The GmMMP2 expressed in and purified from Escherichia coli exhibited an in vitro enzymatic activity in digesting myelin basic protein. All plant metalloproteinases reported so far have no known functions. However, they have been suggested to be involved in extracellular cell matrix degradation during development or senescence. Our investigations demonstrate that the GmMMP2 transcript levels were rapidly increased in compatible and incompatible interactions of soybean tissues with the oomycete pathogen Phytophthora sojae or the bacterial pathogen Pseudomonas syringae pv. glycinea. In agreement with the GmMMP2 activation, a metalloproteinase activity was gradually increased in suspension-cultured cells following the bacterial infection. GmMMP2 was also activated in response to wounding and dehydration. However, GmMMP2 activation did not correlate with the oxidative burst leading to the hypersensitive response cell death or the tissue senescence progress that involves programmed cell death. Our investigations suggest that GmMMP2 may be involved in a novel defense response of soybean against pathogenic infections.
Yang, Liang; Ji, Wei; Zhu, Yanming; Gao, Peng; Li, Yong; Cai, Hua; Bai, Xi; Guo, Dianjing
Calcium/calmodulin-dependent kinases play vital roles in protein phosphorylation in eukaryotes, yet little is known about the phosphorylation process of calcium/calmodulin-dependent protein kinase and its role in stress signal transduction in plants. A novel plant-specific calcium-dependent calmodulin-binding receptor-like kinase (GsCBRLK) has been isolated from Glycine soja. A subcellular localization study using GFP fusion protein indicated that GsCBRLK is localized in the plasma membrane. Binding assays demonstrated that calmodulin binds to GsCBRLK with an affinity of 25.9 nM in a calcium-dependent manner and the binding motif lies between amino acids 147 to169 within subdomain II of the kinase domain. GsCBRLK undergoes autophosphorylation and Myelin Basis Protein phosphorylation in the presence of calcium. It was also found that calcium/calmodulin positively regulates GsCBRLK kinase activity through direct interaction between the calmodulin-binding domain and calmodulin. So, it is likely that GsCBRLK responds to an environmental stimulus in two ways: by increasing the protein expression level and by regulating its kinase activity through the calcium/calmodulin complex. Furthermore, cold, salinity, drought, and ABA stress induce GsCBRLK gene transcripts. Over-expression of GsCBRLK in transgenic Arabidopsis resulted in enhanced plant tolerance to high salinity and ABA and increased the expression pattern of a number of stress gene markers in response to ABA and high salt. These results identify GsCBRLK as a molecular link between the stress- and ABA-induced calcium/calmodulin signal and gene expression in plant cells.
Jang, Seong-Jin; Sato, Masako; Sato, Kei; Jitsuyama, Yutaka; Fujino, Kaien; Mori, Haruhide; Takahashi, Ryoji; Benitez, Eduardo R.; Liu, Baohui; Yamada, Tetsuya; Abe, Jun
Physical dormancy, a structural feature of the seed coat known as hard seededness, is an important characteristic for adaptation of plants against unstable and unpredictable environments. To dissect the molecular basis of qHS1, a quantitative trait locus for hard seededness in soybean (Glycine max (L) Merr.), we developed a near-isogenic line (NIL) of a permeable (soft-seeded) cultivar, Tachinagaha, containing a hard-seed allele from wild soybean (G. soja) introduced by successive backcrossings. The hard-seed allele made the seed coat of Tachinagaha more rigid by increasing the amount of β-1,4-glucans in the outer layer of palisade cells of the seed coat on the dorsal side of seeds, known to be a point of entrance of water. Fine-mapping and subsequent expression and sequencing analyses revealed that qHS1 encodes an endo-1,4-β-glucanase. A single-nucleotide polymorphism (SNP) introduced an amino acid substitution in a substrate-binding cleft of the enzyme, possibly reducing or eliminating its affinity for substrates in permeable cultivars. Introduction of the genomic region of qHS1 from the impermeable (hard-seeded) NIL into the permeable cultivar Kariyutaka resulted in accumulation of β-1,4-glucan in the outer layer of palisade cells and production of hard seeds. The SNP allele found in the NIL was further associated with the occurrence of hard seeds in soybean cultivars of various origins. The findings of this and previous studies may indicate that qHS1 is involved in the accumulation of β-1,4-glucan derivatives such as xyloglucan and/or β-(1,3)(1,4)-glucan that reinforce the impermeability of seed coats in soybean. PMID:26039079
Hafez, Mohamed; Burger, Gertraud; Steinberg, Sergey V.; Lang, Franz
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 tRNAAla-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 α-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
Oborník, Miroslav; Green, Beverley R
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.
Morris, Paul Francis; Schlosser, Laura Rose; Onasch, Katherine Diane; Wittenschlaeger, Tom; Austin, Ryan; Provart, Nicholas
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
Morris, Paul Francis; Schlosser, Laura Rose; Onasch, Katherine Diane; Wittenschlaeger, Tom; Austin, Ryan; Provart, Nicholas
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
Guo, Juan; Wang, Yunsheng; Song, Chi; Zhou, Jianfeng; Qiu, Lijuan; Huang, Hongwen; Wang, Ying
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 approximately 2 % of the total number of ancestral wild soybeans. Wild soybeans in Asia, especially in
Yu, Xiumei; Cloutier, Sylvie; Tambong, James T; Bromfield, Eden S P
Sixteen strains of symbiotic bacteria from root nodules of Glycine max grown in Ottawa, Canada, were previously characterized and placed in a novel group within the genus Bradyrhizobium. To verify their taxonomic status, these strains were further characterized using a polyphasic approach. All strains possessed identical 16S rRNA gene sequences that were 99.79 % similar to the closest relative, Bradyrhizobium liaoningense LMG 18230(T). Phylogenetic analysis of concatenated atpD, glnII, recA, gyrB, rpoB and dnaK genes divided the 16 strains into three multilocus sequence types that were placed in a highly supported lineage distinct from named species of the genus Bradyrhizobium consistent with results of DNA-DNA hybridization. Based on analysis of symbiosis gene sequences (nodC and nifH), all novel strains were placed in a phylogenetic group with five species of the genus Bradyrhizobium that nodulate soybeans. The combination of phenotypic characteristics from several tests including carbon and nitrogen source utilization and antibiotic resistance could be used to differentiate representative strains from recognized species of the genus Bradyrhizobium. Novel strain OO99(T) elicits effective nodules on Glycine max, Glycine soja and Macroptilium atropurpureum, partially effective nodules on Desmodium canadense and Vigna unguiculata, and ineffective nodules on Amphicarpaea bracteata and Phaseolus vulgaris. Based on the data presented, we conclude that our strains represent a novel species for which the name Bradyrhizobium ottawaense sp. nov. is proposed, with OO99(T) ( = LMG 26739(T) = HAMBI 3284(T)) as the type strain. The DNA G+C content is 62.6 mol%.
Guan, Rongxia; Qu, Yue; Guo, Yong; Yu, Lili; Liu, Ying; Jiang, Jinghan; Chen, Jiangang; Ren, Yulong; Liu, Guangyu; Tian, Lei; Jin, Longguo; Liu, Zhangxiong; Hong, Huilong; Chang, Ruzhen; Gilliham, Matthew; Qiu, Lijuan
The identification of genes that improve the salt tolerance of crops is essential for the effective utilization of saline soils for agriculture. Here, we use fine mapping in a soybean (Glycine max (L.) Merr.) population derived from the commercial cultivars Tiefeng 8 and 85-140 to identify GmSALT3 (salt tolerance-associated gene on chromosome 3), a dominant gene associated with limiting the accumulation of sodium ions (Na+) in shoots and a substantial enhancement in salt tolerance in soybean. GmSALT3 encodes a protein from the cation/H+ exchanger family that we localized to the endoplasmic reticulum and which is preferentially expressed in the salt-tolerant parent Tiefeng 8 within root cells associated with phloem and xylem. We identified in the salt-sensitive parent, 85-140, a 3.78-kb copia retrotransposon insertion in exon 3 of Gmsalt3 that truncates the transcript. By sequencing 31 soybean landraces and 22 wild soybean (Glycine soja) a total of nine haplotypes including two salt-tolerant haplotypes and seven salt-sensitive haplotypes were identified. By analysing the distribution of haplotypes among 172 Chinese soybean landraces and 57 wild soybean we found that haplotype 1 (H1, found in Tiefeng 8) was strongly associated with salt tolerance and is likely to be the ancestral allele. Alleles H2-H6, H8 and H9, which do not confer salinity tolerance, were acquired more recently. H1, unlike other alleles, has a wide geographical range including saline areas, which indicates it is maintained when required but its potent stress tolerance can be lost during natural selection and domestication. GmSALT3 is a gene associated with salt tolerance with great potential for soybean improvement.
Hraber, Peter T; Weller, Jennifer W
Background Most organisms have developed ways to recognize and interact with other species. Symbiotic interactions range from pathogenic to mutualistic. Some molecular mechanisms of interspecific interaction are well understood, but many remain to be discovered. Expressed sequence tags (ESTs) from cultures of interacting symbionts can help identify transcripts that regulate symbiosis, but present a unique challenge for functional analysis. Given a sequence expressed in an interaction between two symbionts, the challenge is to determine from which organism the transcript originated. For high-throughput sequencing from interaction cultures, a reliable computational approach is needed. Previous investigations into GC nucleotide content and comparative similarity searching provide provisional solutions, but a comparative lexical analysis, which uses a likelihood-ratio test of hexamer counts, is more powerful. Results Validation with genes whose origin and function are known yielded 94% accuracy. Microbial (non-plant) transcripts comprised 75% of a Phytophthora sojae-infected soybean (Glycine max cv Harasoy) library, contrasted with 15% or less in root tissue libraries of Medicago truncatula from axenic, Phytophthora medicaginis-infected, mycorrhizal, and rhizobacterial treatments. Mycorrhizal libraries contained about 23% microbial transcripts; an axenic plant library contained a similar proportion of putative microbial transcripts. Conclusions Comparative lexical analysis offers numerous advantages over alternative approaches. Many of the transcripts isolated from mixed cultures were of unknown function, suggesting specificity to symbiotic metabolism and therefore candidates likely to be interesting for further functional investigation. Future investigations will determine whether the abundance of non-plant transcripts in a pure plant library indicates procedural artifacts, horizontally transferred genes, or other phenomena. PMID:11574056
Behnam, S; Ahmadzadeh, M; Sharifi Tehrani, A; Hedjaroude, Gh A; Farzaneh, M
Sclerotinia sclerotiorum is an important pathogen on canola. Due to the public concern over pesticide use, alternative methods of disease control, such as biological control, should be considered. Several bacterial strains were isolated from canola and soja plants. Inhibition of S. sclerotiorum by bacterial strains in vitro was assayed on PDA medium in dual culture test. Eight Pseudomonas sp. strains (PB-3, PB-4, PB-5, PB-6, PB-7, PB-8, PB-10 and PB-11) caused inhibition zone against 5. sclerotiorum hyphal growth. The biocontrol potential of the bacteria was tested in a plant assay. Disease suppression was investigated using a petal inoculation technique. Canola petals were pretreated with bacteria, and then inoculated with 5. sclerotiorum ascospores 24 h later. Greenhouse experiment showed that application of Pseudomonas sp. strains (1 x 10(8) cfu ml(-1)) effectively suppressed S. sclerotiorum (1 x 10(5) ascospores ml(-1)) on petals and all of them achieved significant (P<0.01) disease suppression. Fourteen days after inoculation, strain PB-3 had 88/7% disease control and strain PB-4 had 69/9% disease control. Result from all studies indicates PB-3 to be effective biocontrol against S. sclerotiorum of canola. PB-3, PB-4, PB-7, PB-8, PB-10 and PB-11 were identified as Pseudomonas fluorescens biovar III. PB-5 and PB-6 was identified as Pseudomonas fluorescens biovar II. Strains PB-3, PB-4, PB-6, PB-10 and PB-11 produced protease and HCN. Strain PB-5 produce protease; no HCN.
Sun, Mingzhe; Sun, Xiaoli; Zhao, Yang; Zhao, Chaoyue; Duanmu, Huizi; Yu, Yang; Ji, Wei; Zhu, Yanming
So far, it has been suggested that phosphoenolpyruvate carboxylases (PEPCs) and PEPC kinases (PPCKs) fulfill several important non-photosynthetic functions. However, the biological functions of soybean PPCKs, especially in alkali stress response, are not yet well known. In previous studies, we constructed a Glycine soja transcriptional profile, and identified three PPCK genes (GsPPCK1, GsPPCK2 and GsPPCK3) as potential alkali stress responsive genes. In this study, we confirmed the induced expression of GsPPCK3 under alkali stress and investigated its tissue expression specificity by using quantitative real-time PCR analysis. Then we ectopically expressed GsPPCK3 in Medicago sativa and found that GsPPCK3 overexpression improved plant alkali tolerance, as evidenced by lower levels of relative ion leakage and MDA content and higher levels of chlorophyll content and root activity. In this respect, we further co-transformed the GsPPCK3 and SCMRP genes into alfalfa, and demonstrated the increased alkali tolerance of GsPPCK3-SCMRP transgenic lines. Further investigation revealed that GsPPCK3-SCMRP co-overexpression promoted the PEPC activity, net photosynthetic rate and citric acid content of transgenic alfalfa under alkali stress. Moreover, we also observed the up-regulated expression of PEPC, CS (citrate synthase), H(+)-ATPase and NADP-ME genes in GsPPCK3-SCMRP transgenic alfalfa under alkali stress. As expected, we demonstrated that GsPPCK3-SCMRP transgenic lines displayed higher methionine content than wild type alfalfa. Taken together, results presented in this study supported the positive role of GsPPCK3 in plant response to alkali stress, and provided an effective way to simultaneously improve plant alkaline tolerance and methionine content, at least in legume crops.
Tang, Lili; Cai, Hua; Ji, Wei; Luo, Xiao; Wang, Zhenyu; Wu, Jing; Wang, Xuedong; Cui, Lin; Wang, Yang; Zhu, Yanming; Bai, Xi
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.
Van Holle, Sofie; Smagghe, Guy; Van Damme, Els J. M.
Plants have evolved a sophisticated immune system that allows them to recognize invading pathogens by specialized receptors. Carbohydrate-binding proteins or lectins are part of this immune system and especially the lectins that reside in the nucleocytoplasmic compartment are known to be implicated in biotic and abiotic stress responses. The class of Nictaba-like lectins (NLL) groups all proteins with homology to the tobacco (Nicotiana tabacum) lectin, known as a stress-inducible lectin. Here we focus on two Nictaba homologs from soybean (Glycine max), referred to as GmNLL1 and GmNLL2. Confocal laser scanning microscopy of fusion constructs with the green fluorescent protein either transiently expressed in Nicotiana benthamiana leaves or stably transformed in tobacco BY-2 suspension cells revealed a nucleocytoplasmic localization for the GmNLLs under study. RT-qPCR analysis of the transcript levels for the Nictaba-like lectins in soybean demonstrated that the genes are expressed in several tissues throughout the development of the plant. Furthermore, it was shown that salt treatment, Phytophthora sojae infection and Aphis glycines infestation trigger the expression of particular NLL genes. Stress experiments with Arabidopsis lines overexpressing the NLLs from soybean yielded an enhanced tolerance of the plant toward bacterial infection (Pseudomonas syringae), insect infestation (Myzus persicae) and salinity. Our data showed a better performance of the transgenic lines compared to wild type plants, indicating that the NLLs from soybean are implicated in the stress response. These data can help to further elucidate the physiological importance of the Nictaba-like lectins from soybean, which can ultimately lead to the design of crop plants with a better tolerance to changing environmental conditions. PMID:27826309
Garnica, Diana P; Pinzón, Andrés M; Quesada-Ocampo, Lina M; Bernal, Adriana J; Barreto, Emiliano; Grünwald, Niklaus J; Restrepo, Silvia
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. PMID:17007642
Groisman, Pavel; Soja, Amber J.
A brief overview of the ongoing climatic and environmental changes in Northern Eurasia serves as an editorial introduction to this, the second, special Northern Eurasia Earth Science Partnership Initiative (NEESPI) focus issue of Environmental Research Letters. Climatic changes in Northern Eurasia over the last hundred years are reflected in numerous atmospheric and terrestrial variables. Many of these are noticeably significant above the confidence level for 'weather' or other (fire regime, ecosystem change) noise and thus should be further investigated in order to adapt to their impacts. In this focus issue, we introduce assorted studies of different aspects of contemporary change in Northern Eurasia. Most of these have been presented at one of the NEESPI workshops (for more information see neespi.org) and/or American Geophysical Union and European Geosciences Union NEESPI open sessions during the past year. These studies are diverse, representing the diversity of climates and ecosystems across Northern Eurasia. Some of these are focused on smaller spatial scales and/or address only specific aspects of the global change implications across the subcontinent. But the feeling (and observational evidence) that these changes have already been quite rapid and can have global implications inspires us to bring this suite of papers to the readers' attention. See the PDF for the full text of the editorial. Focus on Climatic and Environmental Change in Northern Eurasia Contents Preface Northern Eurasia Earth Science Partnership Initiative Pavel Groisman and Amber J Soja Editorial Siberia integrated regional study: Multidisciplinary investigations of interrelation between Siberia environment dynamics and global climate change E P Gordov and E A Vaganov Studies of the energy and water cycles in Northern Eurasia Comparison and evaluation of gridded radiation products across northern Eurasia T J Troy and E F Wood Reanalysis data underestimate significant changes in growing
Wang, Wubin; Liu, Meifeng; Wang, Yufeng; Li, Xuliang; Cheng, Shixuan; Shu, Liping; Yu, Zheping; Kong, Jiejie; Zhao, Tuanjie; Gai, Junyi
Annual wild soybean (Glycine soja Sieb. and Zucc.), the wild progenitor of the cultivated soybean [Glycine max (L.) Merr.], is valuable for improving the later. The construction of a linkage map is crucial for studying the genetic differentiation between these species, but marker density is the main factor limiting the accuracy of such a map. Recent advances in next-generation sequencing technologies allow for the generation of high-density linkage maps. Here, two sets of inter-specific recombinant inbred line populations, named NJIRNP and NJIR4P, composed of 284 and 161 lines, respectively, were generated from the same wild male parent, PI 342618B, and genotyped by restriction-site-associated DNA sequencing. Two linkage maps containing 5,728 and 4,354 bins were constructed based on 89,680 and 80,995 single nucleotide polymorphisms, spanning a total genetic distance of 2204.6 and 2136.7 cM, with an average distance of 0.4 and 0.5 cM between neighboring bins in NJRINP and NJRI4P, respectively. With the two maps, seven well-studied loci, B1 for seed bloom; G and I for seed coat color; E2, E3, qDTF16.1 and two linked FLOWERING LOCUS T for days to flowering, were detected. In addition, two SB and two DTF loci were newly identified in wild soybean. Using two high-density maps, the mapping resolution was enhanced, e.g., G was narrowed to a region of 0.4 Mb on chromosome 1, encompassing 54 gene models, among which only Glyma01g40590 was predicted to be involved in anthocyanin accumulation, and its interaction with I was verified in both populations. In addition, five genes, Glyma16g03030, orthologous to Arabidopsis Phytochrome A (PHYA); Glyma13g28810, Glyma13g29920, and Glyma13g30710 predicted to encode the APETALA 2 (AP2) domain; and Glyma02g00300, involved in response to red or far red light, might be candidate DTF genes. Our results demonstrate that RAD-seq is a cost-effective approach for constructing high-density and high-quality bin maps that can be used to map QTLs
Alam, Intikhab; Hubbard, Simon J; Oliver, Stephen G; Rattray, Magnus
Background Pfam is a general-purpose database of protein domain alignments and profile Hidden Markov Models (HMMs), which is very popular for the annotation of sequence data produced by genome sequencing projects. Pfam provides models that are often very general in terms of the taxa that they cover and it has previously been suggested that such general models may lack some of the specificity or selectivity that would be provided by kingdom-specific models. Results Here we present a general approach to create domain libraries of HMMs for sub-taxa of a kingdom. Taking fungal species as an example, we construct a domain library of HMMs (called Fungal Pfam or FPfam) using sequences from 30 genomes, consisting of 24 species from the ascomycetes group and two basidiomycetes, Ustilago maydis, a fungal pathogen of maize, and the white rot fungus Phanerochaete chrysosporium. In addition, we include the Microsporidion Encephalitozoon cuniculi, an obligate intracellular parasite, and two non-fungal species, the oomycetes Phytophthora sojae and Phytophthora ramorum, both plant pathogens. We evaluate the performance in terms of coverage against the original 30 genomes used in training FPfam and against five more recently sequenced fungal genomes that can be considered as an independent test set. We show that kingdom-specific models such as FPfam can find instances of both novel and well characterized domains, increases overall coverage and detects more domains per sequence with typically higher bitscores than Pfam for the same domain families. An evaluation of the effect of changing E-values on the coverage shows that the performance of FPfam is consistent over the range of E-values applied. Conclusion Kingdom-specific models are shown to provide improved coverage. However, as the models become more specific, some sequences found by Pfam may be missed by the models in FPfam and some of the families represented in the test set are not present in FPfam. Therefore, we recommend
Costa, J; Taliberti, M P
The Secretary of Health of the State of Sao Paulo, Brazil, has implemented a program of supplemental nutrition to pregnant and nursing mothers in all health units. Objective of this work was to verify the acceptability of the program and of the supplemental food itself, Gestal, a protein supplement composed of 15% milk, 5% soja, 35% sugar, and 25% millet flour. Gestal comes in 5 different flavors, in cans or in plastic bags of 500 g. 6 units are given to pregnant mothers, and 9 to nursing mothers; they are to be consumed in 1 month, in a dose of 150 g a day. 54 pregnant and 55 breastfeeding mothers attending the health unit of Ribeirao Preto, Sao Paulo, were observed for a period of 6 months, and a questionnaire was filled in. 36.69% of mothers preferred the marango flavor, 28.44% the coconut flavor, 10.09% the vanilla flavor, 12.84% the natural flavor, and 9.17% the caramel flavor; in addition, 0.91% liked all flavors, and 1.83% disliked all flavors. 28.44% of mothers prepared Gestal with milk, 20.18% with milk and water, and 51.37% with water. Only 33% of mothers consumed the given amount of Gestal by themselves; of these, 47.5% liked it, 17.2% were indifferent to it, and 14.3% disliked it. 30.3% of mothers shared Gestal with their children; of these 37.2% liked it, 20.6% were indifferent, and 61.9% disliked it. 36.7% of mothers did not consume the given amount within a month; of these 15.2% liked the product, 62.1% were indifferent, and 61.9% disliked it. It was obvious from these results that the mothers were not following recommendations for intake of supplemental food. 54 mothers were chosen at random to be visited at home; of the 33 who could be reached 70% had stopped taking Gestal either because the food had dried, or for fear of getting fat, for the death of the child, or even under a doctor's recommendation. The sharing of supplemental food with the family is a frequent reality which is impossible to control, since it implies emotional as well as
Background The core promoter is the region flanking the transcription start site (TSS) that directs formation of the pre-initiation complex. Core promoters have been studied intensively in mammals and yeast, but not in more diverse eukaryotes. Here we investigate core promoters in oomycetes, a group within the Stramenopile kingdom that includes important plant and animal pathogens. Prior studies of a small collection of genes proposed that oomycete core promoters contain a 16 to 19 nt motif bearing an Initiator-like sequence (INR) flanked by a novel sequence named FPR, but this has not been extended to whole-genome analysis. Results We used expectation maximization to find over-represented motifs near TSSs of Phytophthora infestans, the potato blight pathogen. The motifs corresponded to INR, FPR, and a new element found about 25 nt downstream of the TSS called DPEP. TATA boxes were not detected. Assays of DPEP function by mutagenesis were consistent with its role as a core motif. Genome-wide searches found a well-conserved combined INR+FPR in only about 13% of genes after correcting for false discovery, which contradicted prior reports that INR and FPR are found together in most genes. INR or FPR were found alone near TSSs in 18% and 7% of genes, respectively. Promoters lacking the motifs had pyrimidine-rich regions near the TSS. The combined INR+FPR motif was linked to higher than average mRNA levels, developmentally-regulated transcription, and functions related to plant infection, while DPEP and FPR were over-represented in constitutively-expressed genes. The INR, FPR, and combined INR+FPR motifs were detected in other oomycetes including Hyaloperonospora arabidopsidis, Phytophthora sojae, Pythium ultimum, and Saprolegnia parasitica, while DPEP was found in all but S. parasitica. Only INR seemed present in a non-oomycete stramenopile. Conclusions The absence of a TATA box and presence of novel motifs show that the oomycete core promoter is diverged from that of
Maxelon, Michael; Mancktelow, Neil S.
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
Gurgurewicz, J.; Kostylew, J.
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:  Komatsu G. (2003) Lunar Planet. Sci. XXXIV, abstract 1314.  Soja C. M. and White B. (2006) Geol. Soc. Amer., 38, 90.  Parfenov L. M. et al. (2002) Northeast Asia geodynamics map.
Stackhouse, P. W.; Soja, A. J.; Zhang, T.; Mikovitz, J. C.
In terms of global change, boreal regions are particularly important, because significant warming and change are already evident and significant future warming is predicted. Mean global air temperature has increased by 0.74°C in the last century, and temperatures are predicted to increase by 1.8°C to 4°C by 2090, depending on the Inter-governmental Panel on Climate Change (IPCC) scenario. Some of the greatest temperature increases are currently found in the Northern Eurasian winter and spring, which has led to longer growing seasons, increased potential evapotranspiration and extreme fire weather [Groisman et al., 2007]. In the Siberian Sayan, winter temperatures have already exceeded a 2090 Hadley Centre scenario (HadCM3GGa1) [Soja et al., 2007]. There is evidence of climate-induced change across the circumboreal in terms of increased infestations, alterations in vegetation and increased fire regimes (area burned, fire frequency, severity and number of extreme fire seasons). In this paper, we analyzed long-term surface radiation data sets from the NASA/GEWEX (Global Energy and Water Exchanges) Surface Radiation Budget data products, CERES Surface EBAF and SYN data products and also the available surface radiation measurements in the region. First, we show that during overlap years SRB and CERES data products agree very well in terms of anomalies and we'll use this fact to evaluate 30 years of satellite based estimates of the variability of downwelling SW parameters first corresponding to locations of surface measurements and then for the region as a whole. We also show the observed variability of other SW components such as the net SW and the albedo. Next we assess the variability of the downward and LW fluxes over time and compare these to variability observed in the surface temperature and other meteorological measurements. We assess anomalies on various spatial scales. Finally, we assess the correlation of this variability in specific locations to known fire
Sekhar, Aswin; ASHER, DAVID
Introduction: The close encounter of Comet C/2013 A1 (Siding Spring) with Mars on 2014 Oct 19 at 1830h (UT) generated a lot of interest and modelling work    in the solar system community. A recent (on 2014 Nov 7) press release from NASA implied that a meteor shower was detected on Mars by their space instruments some hours after the comet-Mars close encounter. Various work    has suggested that very specific meteoroid sizes and ejection conditions may be required to produce meteor phenomena at Mars at the given times.Stream dynamics: Meteoroid stream modelling and their orbital geometry calculations have gained high precision over the years. In this work, we compute in detail the structure of the cloud of meteoroids released by C/2013 A1, showing its dependence on heliocentric ejection distances, 3-dimensional ejection velocities, and particle sizes. Our calculations using numerical integrator MERCURY, , incorporating radiation pressure, , show that ejection of particles at large heliocentric distances (about 7 au to 13 au) from C/2013 A1 could lead to evolution of a dense meteoroid cloud which intersects Mars a few hours after the comet-Mars close encounter. Hence this detection of a meteor shower on Mars by space instruments is an indirect confirmation of cometary activity at large distances which has rarely been observed directly by telescopes so far. Furthermore it shows that comprehensive threat estimation needs to be done for satellites orbiting the Earth when dynamically new comets come very close to the Earth in future.References: Vaubaillon J., Macquet L., Soja R. 2014. MNRAS. 439: 3294. Moorhead A. V., Wiegert P. A., Cooke W. J. 2014. Icarus. 231:13. Ye Q.-Z., Hui M.-T., 2014, ApJ, 787: 115. Farnocchia D. et al. 2014. ApJL. 790: 114. Kelley M. S. P. et al. 2014, ApJL, 792: 16. Tricarico P. et al., 2014, ApJL, 787: 35. Chambers J. E. 1999. MNRAS. 304: 793. Burns J. A, Lamy P. L., Soter S. 1979. Icarus. 40: 1.
Wang, Xiaobo; Li, Yinhui; Zhang, Haowei; Sun, Genlou; Zhang, Wenming; Qiu, Lijuan
Seed-size/weight traits, controlled by multiple genes in soybean, play an important role in determining seed yield. However, the molecular mechanisms controlling the seed size and weight in soybean remain unclear. In Arabidopsis, P450/CYP78A gene family has been proved extremely relevant to seed size (such as AtCYP78A5, AtCYP78A6 and AtCYP78A9). We found that a soybean GmCYP78A10 gene underwent artificial selection during soybean breeding. The GmCYP78A10a allele mainly distributed in wild soybean (Glycine soja), but has been eliminated in the cultivars during early stage of soybean breeding, while the GmCYP78A10b allele has been accumulated and become the predominant allele in cultivated soybean (G. max). ANOVA analysis showed that the mean seed weight, seed width and seed thickness of soybean varieties with GmCYP78A10b allele was significantly heavier/bigger than those with GmCYP78A10a allele (P < 0.01). The allele could explain 7.2 % variation in seed weight. The pod number of the soybeans with GmCYP78A10b allele significantly decreased compared to those with GmCYP78A10a allele (P < 0.01, R(2) = 5.8 %), while other agronomic traits including seed weight/plant were not significantly affected by these two alleles. We speculated that during the early stage of soybean breeding, breeders selected big seed carrying GmCYP78A10b allele, but lowered pod number simultaneously. Overall, the selection did not cause the significantly change in soybean seed yield. Our results suggests that the soybean GmCYP78A10 gene may have a similar function to those genes belonging to P450/CYP78A subfamily in Arabidopsis and provides new information for the genetic control of seed size in soybean.
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.
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. PMID:23946659
Vaquerizo Alonso, Clara; Mesejo, Alfonso; Acosta Escribano, José; Ruiz Santana, Sergio
y en glucosa, el 42% con SMOF (Soja, MCT, Oliva, Pescado) y el 33% con EBAO (Emulsiones Basadas en Aceite de Oliva) como emulsión lipídica. El 92% adiciona glutamina. Un 60% considera que la nueva fórmula puede estar indicada en sepsis, trauma, quemados y FMO (Fracaso Multiorgánico) y un 30% la utilizaría de rutina al ingreso. Un 40% considera que disminuye las necesidades de insulina, 50% un mejor manejo del volumen y un 60% destaca la relación proteínas/volumen. Evolutivamente, los pacientes con fórmula específica tienen menor afectación hepática, mayor aporte proteico y menor aporte de volumen, sin diferencias significativas, y precisaron menos dosis de insulina (p = 0,07). Conclusiones: En las doce UCIs seleccionadas, los patrones de práctica clínica en el manejo de la nutrición parenteral se adaptan a las recomendaciones de sociedades científicas como la Sociedad Española de Medicina Intensiva, Crítica y Unidades Coronarias (SEMICYUC), Sociedad Española de Nutrición Parenteral y Enteral (SENPE) y European Society for Clinical Nutrition and Metabolism (ESPEN). Las nuevas fórmulas diseñadas para el paciente grave pueden tener indicaciones en estas patologías.
Gordov, E. P.; Vaganov, E. A.
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