ABSTRACT
Stearoyl-ACP Δ⁹ desaturase (SAD) catalyzes the synthesis of monounsaturated oleic acid or palmitoleic acid in plastids. SAD is the key enzyme to control the ratio of saturated fatty acids to unsaturated fatty acids in plant cells. In order to analyze the regulation mechanism of soybean oleic acid synthesis, soybean (Glycine max) GmSAD family members were genome-wide identified, and their conserved functional domains and physicochemical properties were also analyzed by bioinformatics tools. The spatiotemporal expression profile of each member of GmSADs was detected by qRT-PCR. The expression vectors of GmSAD5 were constructed. The enzyme activity and biological function of GmSAD5 were examined by Agrobacterium-mediated transient expression in Nicotiana tabacum leaves and genetic transformation of oleic acid-deficient yeast (Saccharomyces cerevisiae) mutant BY4389. Results show that the soybean genome contains five GmSAD family members, all encoding an enzyme protein with diiron center and two conservative histidine enrichment motifs (EENRHG and DEKRHE) specific to SAD enzymes. The active enzyme protein was predicted as a homodimer. Phylogenetic analysis indicated that five GmSADs were divided into two subgroups, which were closely related to AtSSI2 and AtSAD6, respectively. The expression profiles of GmSAD members were significantly different in soybean roots, stems, leaves, flowers, and seeds at different developmental stages. Among them, GmSAD5 expressed highly in the middle and late stages of developmental seeds, which coincided with the oil accumulation period. Transient expression of GmSAD5 in tobacco leaves increased the oleic acid and total oil content in leaf tissue by 5.56% and 2.73%, respectively, while stearic acid content was reduced by 2.46%. Functional complementation assay in defective yeast strain BY4389 demonstrated that overexpression of GmSAD5 was able to restore the synthesis of monounsaturated oleic acid, resulting in high oil accumulation. Taken together, soybean GmSAD5 has strong selectivity to stearic acid substrates and can efficiently catalyze the biosynthesis of monounsaturated oleic acid. It lays the foundation for the study of soybean seed oleic acid and total oil accumulation mechanism, providing an excellent target for genetic improvement of oil quality in soybean.
Subject(s)
Fatty Acid Desaturases , Genetics , Metabolism , Gene Expression Profiling , Oleic Acid , Phylogeny , Plant Proteins , Genetics , Seeds , Chemistry , Soybeans , Classification , GeneticsABSTRACT
Enhancing soybean (Glycine max) oil production is crucial to meet the market demand of vegetable oil. Diacylglycerol acyltransferase (DGAT) catalyzes the final acylation reaction of triacylglycerol (TAG) synthesis, acting as one of the rate-limiting enzymes for oil biosynthesis in plant seeds. Here, a cDNA clone VgDGAT1A encoding the DGAT1 protein was isolated from the high oil plant Vernonia galamensis. VgDGAT1A was specifically overexpressed in soybean seeds, and several high-generation transgenic lines (T7) were obtained by continuous selection. qPCR analysis showed that VgDGAT1A was highly expressed in the mid-development stage (30-45 DAF) of the transgenic seeds. Accordingly, the DGAT enzyme activity in the transgenic seeds was increased by 7.8 folds in comparison with the wild-type controls. Seed oil and starch contents were, respectively, increased by 5.1% (Dry weight) and reduced by 2%-3% in the transgenic soybeans. Importantly, protein content was not significantly different between transgenic and control seeds. Seed weight and germination rate of the transgenic lines exhibited no negative effect. Fatty acid profiling demonstrated that antioxidant oleic acid (C18:1Δ9) content in the transgenic seed oil was elevated by 8.2% compared to the control, and correspondingly, easily-oxidized linoleic acid (C18:2Δ9,12) and linolenic acid (C18:3Δ9,12,15) were decreased by 6% and 2% respectively. Taken together, seed-specific overexpression of an exogenous VgDGAT1A gene can break the negative linkage of oil and protein contents in soybean seeds, indicating that engineering of this highly-active DGAT enzyme is an effective strategy to improve oil yield and nutritional value in oilseeds.
ABSTRACT
Aim: Endophytic bacterial population and their diversity in soybean were investigated. Study Design: Endophytic population was assessed during different growth stages of soybean (CV JS 335) viz., vegetative and reproductive stages. Place and Duration of Study: Microbiology Research Laboratory, Department of Microbiology, R. A. Mahavidyalaya, Washim (MS), India, during the cultivation period of June-December 2010. Methodology: Healthy plants of soybean were screened from the different locations of Washim district (M. S., India). Samples represent each growth stage viz., vegetative (V1- V5) and reproductive (R1-R8) were collected. Population densities were expressed as log10 colony forming units (CFU) g-1 fresh weight. The isolates were identified to genus level according to Bergey’s Manual of Determinative Bacteriology on the basis morphological, cultural and biochemical characteristics. Results: The maximum endophytic population was recorded for vegetative stage at V5 and V4 (5.74 and 5.01 log10 CFU g-1 fresh weight) and for reproductive stage at R2, R1 and R3 (5.84, 5.80 and 5.74 log10 CFU g-1 fresh weight). A total of 572 (35.50 %) from vegetative growth stages and 1039 (64.50 %) from reproductive growth stages bacterial isolates were obtained. The endophytic isolates were identified as members belonging to the genera Pseudomonas, Bacillus, Enterobacter, Klebsiella, Acetobacter Burkholderia, Rhizobium and Xanthomonas. Conclusion: As soybean development progresses endophytic population increased. At maturity, the high population density was observed and thereafter the population declined.
ABSTRACT
O Brasil é o segundo maior produtor e exportador de soja (Glycine max L. Merrill) no mundo. A produção é altamente dependente de fatores climáticos, incluindo a temperatura e quantidade de chuva. A soja cultivada no sul do país em 2005 sofreu déficit hídrico causado por temperaturas altas acompanhado por umidade baixa durante o estádio reprodutivo. Pouco se sabe sobre a influência do déficit hídrico na degradação da clorofila e na qualidade dos grãos em geral. Nesse trabalho foram analisadas, numa primeira etapa, as características químicas e bioquímicas de cinco amostras de soja, provenientes de três cultivares que cresceram sob déficit hídrico e que não atendiam aos padrões para comercialização por conter altas quantidades de sementes verdes. Os grãos foram analisados quanto a diversos parâmetros físico-químicos e bioquímicos incluindo a análise dos pigmentos verdes imediatamente após a colheita e após 20 meses de armazenamento. A acidez foi medida adicionalmente após 30 meses de armazenamento. A atividade de água e umidade foram 0,6-0,7 e 8,7 %-11,9 %, respectivamente, e não mudaram durante a estocagem, mas houve um aumento em acidez o que indica atividade de lipases. A atividade da lipoxigenase 1 foi significativamente prejudicada. Imediatamente após a colheita os pigmentos verdes correspondiam a feofitina a, feofitina b e pequenas quantidades de clorofila a e b, e traços de outros derivados da clorofila, em ordem decrescente. Após 20 meses de estocagem quase todos os pigmentos haviam desaparecido. O déficit hídrico provavelmente aumentou a permeabilidade das membranas, o que levou a um aumento do pH e promoveu a transformação das clorofilas para feofitinas. Na segunda etapa do trabalho foi estudado o estágio avançado da degradação natural da clorofila na qual desaparece a coloração esverdeada dos grãos. Esta etapa corresponde à formação de catabólitos incolores (NCC), mas existem controvérsias se essas substâncias são os produtos finais da degradação...
Brazil is the second largest soybean (Glycine max L. Merrill) producer and exporter in the world. The production depends on climatic factors, like temperature and rain volume. Soybeans cultivated in the south of the country in 2005 suffered drought stress imposed by adverse high ambient temperature, accompanied by low humidity during the reproductive stage. Little information is available regarding drought stress on quality of grains. In this study, firstly, chemical and biochemical characteristics of five soybean samples belonging to three cultivars grown under drought stress and did not meet standards for marketing due to high amounts of green seeds, were evaluated. Grains were analyzed for several physicochemical ad biochemical parameters, including analysis of pigment contents, immediately after harvest and after 20 months of storage at room temperature. Acidity was measured additionally after 30 month of storage. Water activity and humidity were 0.6 - 0.7 and 8.7 % - 11.9 %, respectively, and did not change during storage time, but there was an increase in acidity, which alludes to lipase activity. The activity of lipoxygenase 1 was significantly prejudiced. Immediately after harvest, green pigments corresponded mainly to pheophytin a, followed by pheophytin b, small quantities of chlorophyll b and chlorophyll a and traces of other chlorophyll derivatives. After 20 months of storage almost all green pigments had disappeared. Drought stress probably enhanced membrane permeability, which led to a lower pH and promoted transformation of chlorophylls to pheophytins. In the second part of the study, the advanced stage of natural chlorophyll degradation was investigated, in which the green colour of the seeds disappears. This stage corresponds to the formation of non-coloured chlorophyll catabolites (NCC), but there are controversies if those are the final products. Thus, the formation and degradation of NCC during soybean (Glycine max L. Merrill)...