Species closely related to wheat are important genetic resources for agricultural production,functional genomics studies and wheat improvement.In this study,a wheat gene related to regeneration,TaWOX5,was applied to e...Species closely related to wheat are important genetic resources for agricultural production,functional genomics studies and wheat improvement.In this study,a wheat gene related to regeneration,TaWOX5,was applied to establish the Agrobacterium-mediated transformation systems of Triticum monococcum,hexaploid triticale,and rye(Secale cereale L.)using their immature embryos.Transgenic plants were efficiently generated.During the transformation process,the Agrobacterium infection efficiency was assessed by histochemical staining forβ-glucuronidase(GUS).Finally,the transgenic nature of regenerated plants was verified by polymerase chain reaction(PCR)-based genotyping for the presence of the GUS and bialaphos resistance(bar)genes,histochemical staining for GUS protein,and the QuickStix strip assay for bar protein.The transformation efficiency of T.monococcum genotype PI428182 was 94.4%;the efficiencies of four hexaploid triticale genotypes Lin456,ZS3297,ZS1257,and ZS3224 were 52.1,41.2,19.4,and 16.0%,respectively;and the transformation efficiency of rye cultivar Lanzhou Heimai was 7.8%.Fluorescence in situ hybridization(FISH)and genomic in situ hybridization(GISH)analyses indicated that the GUS transgenes were integrated into the distal or near centromere(proximal)regions of the chromosomes in transgenic T.monococcum and hexaploid triticale plants.In the transgenic hexaploid triticale plants,the foreign DNA fragment was randomly integrated into the AABB and RR genomes.Furthermore,the transgene was almost stably inherited in the next generation by Mendel’s law.The findings in this study will promote the genetic improvement of the three plant species for grain or forage production and the improvement of cereal species including wheat for functional genomics studies.展开更多
Wheat seed development is an important physiological process of seed maturation and directly affects wheat yield and quality. In this study, we performed dynamic transcriptome microarray analysis of an elite Chinese b...Wheat seed development is an important physiological process of seed maturation and directly affects wheat yield and quality. In this study, we performed dynamic transcriptome microarray analysis of an elite Chinese bread wheat cultivar(Jimai 20) during grain development using the Gene Chip Wheat Genome Array. Grain morphology and scanning electron microscope observations showed that the period of 11–15 days post-anthesis(DPA) was a key stage for the synthesis and accumulation of seed starch. Genome-wide transcriptional profiling and significance analysis of microarrays revealed that the period from 11 to 15 DPA was more important than the 15–20 DPA stage for the synthesis and accumulation of nutritive reserves.Series test of cluster analysis of differential genes revealed five statistically significant gene expression profiles. Gene ontology annotation and enrichment analysis gave further information about differentially expressed genes, and Map Man analysis revealed expression changes within functional groups during seed development. Metabolic pathway network analysis showed that major and minor metabolic pathways regulate one another to ensure regular seed development and nutritive reserve accumulation. We performed gene co-expression network analysis to identify genes that play vital roles in seed development and identified several key genes involved in important metabolic pathways. The transcriptional expression of eight key genes involved in starch and protein synthesis and stress defense was further validated by q RT-PCR. Our results provide new insight into the molecular mechanisms of wheat seed development and the determinants of yield and quality.展开更多
The content and composition of wheat storage proteins are the major determinants of dough rheological properties and breadmaking quality and are influenced by cultivation conditions.This study aimed to investigate the...The content and composition of wheat storage proteins are the major determinants of dough rheological properties and breadmaking quality and are influenced by cultivation conditions.This study aimed to investigate the effects of water deficit and high N-fertilizer application on wheat storage protein synthesis,gluten secondary structure,and breadmaking quality.Reverse-phase ultrahigh-performance liquid chromatography analysis showed that storage protein and gluten macropolymer accumulation was promoted under both independent applications and a combination of water-deficit and high N-fertilizer treatments.Fourier-transform infrared spectroscopy showed that water deficit and high N-fertilizer treatments generally improved protein secondary structure formation and lipid accumulation,and reduced flour moisture.In particular,high N-fertilizer application increasedβ-sheet content by 10.4%and the combination of water-deficit and high N-fertilizer treatments increased random coil content by 7.6%.These changes in gluten content and secondary structure led to improved dough rheological properties and breadmaking quality,including superior loaf internal structure,volume,and score.Our results demonstrate that moderately high N-fertilizer application under drought conditions can improve gluten accumulation,gluten secondary structure formation,and baking quality.展开更多
Nitrogen(N),a macronutrient essential for plant growth and development,is needed for biosynthesis of protein and starch,which affect grain yield and quality.Application of high-N fertilizer increases plant growth,grai...Nitrogen(N),a macronutrient essential for plant growth and development,is needed for biosynthesis of protein and starch,which affect grain yield and quality.Application of high-N fertilizer increases plant growth,grain yield,and flour quality.In this study,we performed the first comparative analysis of gliadin and glutenin subproteomes during kernel development in the elite Chinese wheat cultivar Zhongmai 175 under high-N conditions by reversed-phase ultra-performance liquid chromatography and twodimensional difference gel electrophoresis(2D-DIGE).Application of high-N fertilizer led to significant increases in gluten macropolymer content,total gliadin and glutenin content,and the accumulation of individual storage protein components.Of 126 differentially accumulated proteins(DAPs)induced by high-N conditions,24 gliadins,12 high-molecularweight glutenins,and 27 low-molecular-weight glutenins were significantly upregulated.DAPs during five kernel developmental stages displayed multiple patterns of accumulation.In particular,gliadins and glutenins showed respectively five and six accumulation patterns.The accumulation of storage proteins under high-N conditions may lead to improved dough properties and bread quality.展开更多
Reserve starch of cereal crop accounts for about 70%of grain endosperm and acts as an important human carbohydrate resource worldwide.Wheat reserve starch is synthesized by enzymatic machinery in endosperm cells.To id...Reserve starch of cereal crop accounts for about 70%of grain endosperm and acts as an important human carbohydrate resource worldwide.Wheat reserve starch is synthesized by enzymatic machinery in endosperm cells.To identify genes involved in starch biosynthesis,we constructed 30 RNA-Seq libraries of 10 endosperm-development periods and performed expression and localization analyses.Of 166 endosperm-expressed homologs of starch biosynthesis-related genes,74 showed expression correlated with reserve starch accumulation,including 26 with expected subcellular distribution and higher expression than their isoforms.The key proteins SUS3,UGP1,cAGPase,and Bt1-3 formed the main metabolic pathway and contributed the major substrates for starch processing in amyloplasts.Important isoforms,key pathway proteins,and the main carbon flux toward starch formation in the reserve starch biosynthesis pathway were identified.Based on a coexpression analysis,a library of 425 transcription factors was produced to screen for common regulators.TaMYB44 had features of transcription factors and bound to TaSUT1,TaSSIIIa,TaBEIIa,TaISA1,and TaBEIIb promoters in yeast,suggesting that the gene is a pathway regulator.This study sheds light on understanding the mechanism of reserve starch biosynthesis and will be helpful for increasing starch content in wheat endosperm via biotechnological strategies.展开更多
基金supported by grants from the National Natural Science Foundation of China (31971946)the Technology Innovation Program of the Chinese Academy of Agricultural Sciences, China (2060302-2-23 and ASTIP-2060302-2-19).
文摘Species closely related to wheat are important genetic resources for agricultural production,functional genomics studies and wheat improvement.In this study,a wheat gene related to regeneration,TaWOX5,was applied to establish the Agrobacterium-mediated transformation systems of Triticum monococcum,hexaploid triticale,and rye(Secale cereale L.)using their immature embryos.Transgenic plants were efficiently generated.During the transformation process,the Agrobacterium infection efficiency was assessed by histochemical staining forβ-glucuronidase(GUS).Finally,the transgenic nature of regenerated plants was verified by polymerase chain reaction(PCR)-based genotyping for the presence of the GUS and bialaphos resistance(bar)genes,histochemical staining for GUS protein,and the QuickStix strip assay for bar protein.The transformation efficiency of T.monococcum genotype PI428182 was 94.4%;the efficiencies of four hexaploid triticale genotypes Lin456,ZS3297,ZS1257,and ZS3224 were 52.1,41.2,19.4,and 16.0%,respectively;and the transformation efficiency of rye cultivar Lanzhou Heimai was 7.8%.Fluorescence in situ hybridization(FISH)and genomic in situ hybridization(GISH)analyses indicated that the GUS transgenes were integrated into the distal or near centromere(proximal)regions of the chromosomes in transgenic T.monococcum and hexaploid triticale plants.In the transgenic hexaploid triticale plants,the foreign DNA fragment was randomly integrated into the AABB and RR genomes.Furthermore,the transgene was almost stably inherited in the next generation by Mendel’s law.The findings in this study will promote the genetic improvement of the three plant species for grain or forage production and the improvement of cereal species including wheat for functional genomics studies.
基金financially supported by grants from the National Natural Science Foundation of China(31471485)Natural Science Foundation of Beijing Citythe Key Developmental Project of Science and Technology from Beijing Municipal Commission of Education(KZ201410028031)
文摘Wheat seed development is an important physiological process of seed maturation and directly affects wheat yield and quality. In this study, we performed dynamic transcriptome microarray analysis of an elite Chinese bread wheat cultivar(Jimai 20) during grain development using the Gene Chip Wheat Genome Array. Grain morphology and scanning electron microscope observations showed that the period of 11–15 days post-anthesis(DPA) was a key stage for the synthesis and accumulation of seed starch. Genome-wide transcriptional profiling and significance analysis of microarrays revealed that the period from 11 to 15 DPA was more important than the 15–20 DPA stage for the synthesis and accumulation of nutritive reserves.Series test of cluster analysis of differential genes revealed five statistically significant gene expression profiles. Gene ontology annotation and enrichment analysis gave further information about differentially expressed genes, and Map Man analysis revealed expression changes within functional groups during seed development. Metabolic pathway network analysis showed that major and minor metabolic pathways regulate one another to ensure regular seed development and nutritive reserve accumulation. We performed gene co-expression network analysis to identify genes that play vital roles in seed development and identified several key genes involved in important metabolic pathways. The transcriptional expression of eight key genes involved in starch and protein synthesis and stress defense was further validated by q RT-PCR. Our results provide new insight into the molecular mechanisms of wheat seed development and the determinants of yield and quality.
基金financially supported by the National Natural Science Foundation of China(31771773 and 31971931)the National Key Research and Development Program of China(2016YFD0100502)。
文摘The content and composition of wheat storage proteins are the major determinants of dough rheological properties and breadmaking quality and are influenced by cultivation conditions.This study aimed to investigate the effects of water deficit and high N-fertilizer application on wheat storage protein synthesis,gluten secondary structure,and breadmaking quality.Reverse-phase ultrahigh-performance liquid chromatography analysis showed that storage protein and gluten macropolymer accumulation was promoted under both independent applications and a combination of water-deficit and high N-fertilizer treatments.Fourier-transform infrared spectroscopy showed that water deficit and high N-fertilizer treatments generally improved protein secondary structure formation and lipid accumulation,and reduced flour moisture.In particular,high N-fertilizer application increasedβ-sheet content by 10.4%and the combination of water-deficit and high N-fertilizer treatments increased random coil content by 7.6%.These changes in gluten content and secondary structure led to improved dough rheological properties and breadmaking quality,including superior loaf internal structure,volume,and score.Our results demonstrate that moderately high N-fertilizer application under drought conditions can improve gluten accumulation,gluten secondary structure formation,and baking quality.
基金financially supported by the National Key Research and Development Program of China(2016YFD0100502)the National Natural Science Foundation of China(31171773)
文摘Nitrogen(N),a macronutrient essential for plant growth and development,is needed for biosynthesis of protein and starch,which affect grain yield and quality.Application of high-N fertilizer increases plant growth,grain yield,and flour quality.In this study,we performed the first comparative analysis of gliadin and glutenin subproteomes during kernel development in the elite Chinese wheat cultivar Zhongmai 175 under high-N conditions by reversed-phase ultra-performance liquid chromatography and twodimensional difference gel electrophoresis(2D-DIGE).Application of high-N fertilizer led to significant increases in gluten macropolymer content,total gliadin and glutenin content,and the accumulation of individual storage protein components.Of 126 differentially accumulated proteins(DAPs)induced by high-N conditions,24 gliadins,12 high-molecularweight glutenins,and 27 low-molecular-weight glutenins were significantly upregulated.DAPs during five kernel developmental stages displayed multiple patterns of accumulation.In particular,gliadins and glutenins showed respectively five and six accumulation patterns.The accumulation of storage proteins under high-N conditions may lead to improved dough properties and bread quality.
基金This research was supported the National Program on R&D of Transgenic Plants(2016ZX08009003-004)National Natural Science Foundation of China(31571652)the Youth Innovative Research Team of Capital Normal University.
文摘Reserve starch of cereal crop accounts for about 70%of grain endosperm and acts as an important human carbohydrate resource worldwide.Wheat reserve starch is synthesized by enzymatic machinery in endosperm cells.To identify genes involved in starch biosynthesis,we constructed 30 RNA-Seq libraries of 10 endosperm-development periods and performed expression and localization analyses.Of 166 endosperm-expressed homologs of starch biosynthesis-related genes,74 showed expression correlated with reserve starch accumulation,including 26 with expected subcellular distribution and higher expression than their isoforms.The key proteins SUS3,UGP1,cAGPase,and Bt1-3 formed the main metabolic pathway and contributed the major substrates for starch processing in amyloplasts.Important isoforms,key pathway proteins,and the main carbon flux toward starch formation in the reserve starch biosynthesis pathway were identified.Based on a coexpression analysis,a library of 425 transcription factors was produced to screen for common regulators.TaMYB44 had features of transcription factors and bound to TaSUT1,TaSSIIIa,TaBEIIa,TaISA1,and TaBEIIb promoters in yeast,suggesting that the gene is a pathway regulator.This study sheds light on understanding the mechanism of reserve starch biosynthesis and will be helpful for increasing starch content in wheat endosperm via biotechnological strategies.