In plants,glycerol-3-phosphate dehydrogenase(GPDH)catalyzes the interconversion of glycerol-3-phosphate(G3P)and dihydroxyacetone phosphate(DHAP)coupled to the reduction/oxidation of the nicotinamide adenine dinucleoti...In plants,glycerol-3-phosphate dehydrogenase(GPDH)catalyzes the interconversion of glycerol-3-phosphate(G3P)and dihydroxyacetone phosphate(DHAP)coupled to the reduction/oxidation of the nicotinamide adenine dinucleotide(NADH)pool,and plays a central role in glycerolipid metabolism and stress response.Previous studies have focused mainly on the NAD+-dependent GPDH isoforms,neglecting the role of flavin adenine dinucleotide(FAD)-dependent GPDHs.We isolated and characterized three mitochondrialtargeted FAD-GPDHs in soybean,of which one isoform(GmGPDH12)showed a significant transcriptional response to NaCl and mannitol treatments,suggesting the existence of a major FAD-GPDH isoform acting in soybean responses to salt and osmotic stress.An enzyme kinetic assay showed that the purified GmGPDH12 protein possessed the capacity to oxidize G3P to DHAP in the presence of FAD.Overexpression and RNA interference of GmGPDH12 in soybean hairy roots resulted in elevated tolerance and sensitivity to salt and osmotic stress,respectively.G3P contents were significantly lower in GmGPDH12-overexpressing hair roots and higher in knockdown hair roots,indicating that GmGPDH12 was essential for G3P catabolism.A significant perturbation in redox status of NADH,ascorbic acid(ASA)and glutathione(GSH)pools was observed in GmGPDH12-knockdown plants under stress conditions.The impaired redox balance was manifested by higher reactive oxygen species generation and consequent cell damage or death;however,overexpressing plants showed the opposite results for these traits.GmGPDH12 overexpression contributed to maintaining constant respiration rates under salt or osmotic stress by regulating mRNA levels of key mitochondrial respiratory enzymes.This study provides new evidence for the roles of mitochondria-localized GmGPDH12 in conferring resistance to salt or osmotic stress by maintaining cellular redox homeostasis,protecting cells and respiration from oxidative injury.展开更多
Phytophthora sojae infection severely impairs soybean production. We previously identified a dirigent protein, Gm DRR1(Glycine max Disease Resistant Response 1), that increases soybean resistance to P.sojae. However, ...Phytophthora sojae infection severely impairs soybean production. We previously identified a dirigent protein, Gm DRR1(Glycine max Disease Resistant Response 1), that increases soybean resistance to P.sojae. However, the molecular basis of Gm DRR1 function remained largely uncharacterized. In the present study, analysis of Gm DRR1-RNAi, Gm DRR1-overexpressing, and CRISPR/Cas9-derived Gmdrr1 mutant lines revealed that Gm DRR1 expression significantly restricted P. sojae growth. Combining coimmunoprecipitation with liquid chromatography–tandem mass spectrometry revealed a Gm DRR1-interacting protein, Gm DRR2, which is homologous to Gm DRR1. An E-coniferyl alcohol coupling assay indicated that Gm DRR1 promotes the synthesis of(+)-pinoresinol, which helps to protect plants from P. sojae. The Gm NAC1(Glyma.05 G025500) transcription factor bound to the Gm DRR1 promoter both in vitro and in vivo to upregulate Gm DRR1 expression. Soybean resistance to P. sojae was increased by overexpression of Gm NAC1. Our findings suggest a novel signaling pathway involving a NAC transcription factor that mediates soybean resistance to P. sojae. Specifically, Gm NAC1 directly induces Gm DRR1 expression to increase resistance of soybean plants to P. sojae.展开更多
There is growing interest in expanding the production of soybean oils(mainly triacylglycerol, or TAG) to meet rising feed demand and address global energy concerns. We report that a plastidlocalized glycerol-3-phospha...There is growing interest in expanding the production of soybean oils(mainly triacylglycerol, or TAG) to meet rising feed demand and address global energy concerns. We report that a plastidlocalized glycerol-3-phosphate dehydrogenase(GPDH), encoded by GmGPDHp1 gene, catalyzes the formation of glycerol-3-phosphate(G3 P), an obligate substrate required for TAG biosynthesis.Overexpression of GmGPDHp1 increases soybean seed oil content with high levels of unsaturated fatty acids(FAs), especially oleic acid(C18:1), without detectably affecting growth or seed protein content or seed weight. Based on the lipidomic analyses, we found that the increase in G3 P content led to an elevated diacylglycerol(DAG) pool, in which the Kennedy pathwayderived DAG was mostly increased, followed by PC-derived DAG, thereby promoting the synthesis of TAG containing relatively high proportion of C18:1. The increased G3 P levels induced several transcriptional alterations of genes involved in the glycerolipid pathways. In particular, genes encoding the enzymes responsible for de novo glycerolipid synthesis were largely upregulated in the transgenic lines, in-line with the identified biochemical phenotype. These results reveal a key role for GmGPDHp1-mediated G3 P metabolism in enhancing TAG synthesis and demonstrate a strategy to modify the FA compositions of soybean oils for improved nutrition and biofuel.展开更多
N^(6)-methyladenosine(m^(6)A)is a reversible epigenetic modification of mRNA and other RNAs that plays a significant role in regulating gene expression and biological processes.However,m^(6)A abundance,dynamics,and tr...N^(6)-methyladenosine(m^(6)A)is a reversible epigenetic modification of mRNA and other RNAs that plays a significant role in regulating gene expression and biological processes.However,m^(6)A abundance,dynamics,and transcriptional regulatory mechanisms remain unexplored in the context of soybean resistance to Meloidogyne incognita.In this study,we performed a comparative analysis of transcriptome-wide m^(6)A and metabolome profiles of soybean root tissues with and without M.incognita infection.Global m^(6)A hypermethylation was widely induced in response to M.incognita infection and was enriched around the 3′end of coding sequences and in 3′UTR regions.There were 2069 significantly modified m^(6)A sites,594 differentially expressed genes,and 103 differentially accumulated metabolites between infected and uninfected roots,including coumestrol,psoralidin,and 2-hydroxyethylphosphonate.Among 101 m^(6)A-modified DEGs,34 genes were hypomethylated and upregulated,and 39 genes were hypermethylated and downregulated,indicating a highly negative correlation between m^(6)A methylation and gene transcript abundance.A number of these m^(6)A-modified DEGs,including WRKY70,ERF60,POD47 and LRR receptor-like serine/threonine-protein kinases,were involved in plant defense responses.Our study provides new insights into the critical role of m^(6)A modification in early soybean responses to M.incognita.展开更多
基金financially supported by National Natural Science Foundation of China(31701449,31971968,31971899,31501332)National Key Research and Development Program of China(2016YFD 0100500,2016YFD0100300,2016YFD0100201-21,JFYS2016ZY03003792-01-21)+7 种基金China Postdoctoral Science Foundation(2019M661243)Postdoctoral Project of Northeast Agricultural University(NEAUBH-19002)EUCLEG(727312,2017YFE0111000)Natural Science Foundation of Heilongjiang Province(QC2017013)Special Financial Aid to Post-doctor Research Fellow in Heilongjiang(LBH-TZ1714)Heilongjiang Academy of Agricultural Sciences Funds(2019YYYF019)International Postdoctoral Exchange Fellowship Program of China Postdoctoral Council(20180004)Heilongjiang Funds for Distinguished Young Scientists(JC2016004,JC2017006)。
文摘In plants,glycerol-3-phosphate dehydrogenase(GPDH)catalyzes the interconversion of glycerol-3-phosphate(G3P)and dihydroxyacetone phosphate(DHAP)coupled to the reduction/oxidation of the nicotinamide adenine dinucleotide(NADH)pool,and plays a central role in glycerolipid metabolism and stress response.Previous studies have focused mainly on the NAD+-dependent GPDH isoforms,neglecting the role of flavin adenine dinucleotide(FAD)-dependent GPDHs.We isolated and characterized three mitochondrialtargeted FAD-GPDHs in soybean,of which one isoform(GmGPDH12)showed a significant transcriptional response to NaCl and mannitol treatments,suggesting the existence of a major FAD-GPDH isoform acting in soybean responses to salt and osmotic stress.An enzyme kinetic assay showed that the purified GmGPDH12 protein possessed the capacity to oxidize G3P to DHAP in the presence of FAD.Overexpression and RNA interference of GmGPDH12 in soybean hairy roots resulted in elevated tolerance and sensitivity to salt and osmotic stress,respectively.G3P contents were significantly lower in GmGPDH12-overexpressing hair roots and higher in knockdown hair roots,indicating that GmGPDH12 was essential for G3P catabolism.A significant perturbation in redox status of NADH,ascorbic acid(ASA)and glutathione(GSH)pools was observed in GmGPDH12-knockdown plants under stress conditions.The impaired redox balance was manifested by higher reactive oxygen species generation and consequent cell damage or death;however,overexpressing plants showed the opposite results for these traits.GmGPDH12 overexpression contributed to maintaining constant respiration rates under salt or osmotic stress by regulating mRNA levels of key mitochondrial respiratory enzymes.This study provides new evidence for the roles of mitochondria-localized GmGPDH12 in conferring resistance to salt or osmotic stress by maintaining cellular redox homeostasis,protecting cells and respiration from oxidative injury.
基金supported by the National Natural Science Foundation of China(U20A2027,32070274,32072014)the Heilongjiang Postdoctoral Science Foundation(LBH-Q16014)。
文摘Phytophthora sojae infection severely impairs soybean production. We previously identified a dirigent protein, Gm DRR1(Glycine max Disease Resistant Response 1), that increases soybean resistance to P.sojae. However, the molecular basis of Gm DRR1 function remained largely uncharacterized. In the present study, analysis of Gm DRR1-RNAi, Gm DRR1-overexpressing, and CRISPR/Cas9-derived Gmdrr1 mutant lines revealed that Gm DRR1 expression significantly restricted P. sojae growth. Combining coimmunoprecipitation with liquid chromatography–tandem mass spectrometry revealed a Gm DRR1-interacting protein, Gm DRR2, which is homologous to Gm DRR1. An E-coniferyl alcohol coupling assay indicated that Gm DRR1 promotes the synthesis of(+)-pinoresinol, which helps to protect plants from P. sojae. The Gm NAC1(Glyma.05 G025500) transcription factor bound to the Gm DRR1 promoter both in vitro and in vivo to upregulate Gm DRR1 expression. Soybean resistance to P. sojae was increased by overexpression of Gm NAC1. Our findings suggest a novel signaling pathway involving a NAC transcription factor that mediates soybean resistance to P. sojae. Specifically, Gm NAC1 directly induces Gm DRR1 expression to increase resistance of soybean plants to P. sojae.
基金financially supported by National Key R&D Program of China(2016YFD0100201,2016YFD0100500,2016YFD0100300)National Natural Science Foundation of China(U20A2027,31971899,31971968,32070274)+7 种基金Hundred-thousand and million project of“Heilongjiang province for engineering and technology science”soybean breeding technology innovation and new cultivar breeding(2019ZX16B01)Natural Science Foundation of Heilongjiang(ZD2020C007)China Postdoctoral Science Foundation(2019M661243)Postdoctoral Project of Northeast Agricultural University(NEAUBH-19002)International Postdoctoral Exchange Fellowship Program of China Postdoctoral Council(20180004)Heilongjiang Funds for Distinguished Young Scientists(JC2016004,JC2017006)Dongnongxue zhe Project(to Chen Qingshan)Backbone of Young Talent Scholar Project(to Qi Zhaoming,18XG01)of Northeast Agricultural University。
文摘There is growing interest in expanding the production of soybean oils(mainly triacylglycerol, or TAG) to meet rising feed demand and address global energy concerns. We report that a plastidlocalized glycerol-3-phosphate dehydrogenase(GPDH), encoded by GmGPDHp1 gene, catalyzes the formation of glycerol-3-phosphate(G3 P), an obligate substrate required for TAG biosynthesis.Overexpression of GmGPDHp1 increases soybean seed oil content with high levels of unsaturated fatty acids(FAs), especially oleic acid(C18:1), without detectably affecting growth or seed protein content or seed weight. Based on the lipidomic analyses, we found that the increase in G3 P content led to an elevated diacylglycerol(DAG) pool, in which the Kennedy pathwayderived DAG was mostly increased, followed by PC-derived DAG, thereby promoting the synthesis of TAG containing relatively high proportion of C18:1. The increased G3 P levels induced several transcriptional alterations of genes involved in the glycerolipid pathways. In particular, genes encoding the enzymes responsible for de novo glycerolipid synthesis were largely upregulated in the transgenic lines, in-line with the identified biochemical phenotype. These results reveal a key role for GmGPDHp1-mediated G3 P metabolism in enhancing TAG synthesis and demonstrate a strategy to modify the FA compositions of soybean oils for improved nutrition and biofuel.
基金supported by the National Natural Science Foundation of China(31901859,31901858)and the Syngenta-NEAU union foundation.
文摘N^(6)-methyladenosine(m^(6)A)is a reversible epigenetic modification of mRNA and other RNAs that plays a significant role in regulating gene expression and biological processes.However,m^(6)A abundance,dynamics,and transcriptional regulatory mechanisms remain unexplored in the context of soybean resistance to Meloidogyne incognita.In this study,we performed a comparative analysis of transcriptome-wide m^(6)A and metabolome profiles of soybean root tissues with and without M.incognita infection.Global m^(6)A hypermethylation was widely induced in response to M.incognita infection and was enriched around the 3′end of coding sequences and in 3′UTR regions.There were 2069 significantly modified m^(6)A sites,594 differentially expressed genes,and 103 differentially accumulated metabolites between infected and uninfected roots,including coumestrol,psoralidin,and 2-hydroxyethylphosphonate.Among 101 m^(6)A-modified DEGs,34 genes were hypomethylated and upregulated,and 39 genes were hypermethylated and downregulated,indicating a highly negative correlation between m^(6)A methylation and gene transcript abundance.A number of these m^(6)A-modified DEGs,including WRKY70,ERF60,POD47 and LRR receptor-like serine/threonine-protein kinases,were involved in plant defense responses.Our study provides new insights into the critical role of m^(6)A modification in early soybean responses to M.incognita.
