Establishment of a highly efficient regeneration system for the mature embryo of wheat will provide a convenient tool for wheat tissue culture and transformation, thereby facilitating the transformation of foreign gen...Establishment of a highly efficient regeneration system for the mature embryo of wheat will provide a convenient tool for wheat tissue culture and transformation, thereby facilitating the transformation of foreign genes into wheat. By using the mature embryos derived from 20 different wheat lines including Shi 4185, Yumai 66, Lunxuan 987, CB037, Yangmai 6, Xinchun 9, Bobwhite, Han 6172, Zheng 9023, Jimai 20, Ningchun 4, and Jing 411, the effects of some factors including inoculation methods, initiating culture media, organic additives, antioxidants, and auxins on the regeneration from the explants were evaluated. The results indicated that the scraping embryo culture was better than the whole embryo culture, the Aa medium was better than the SD2 medium and dicamba was better than 2,4-D in increasing the regeneration frequency. An Adi medium was established in this study by adding silver nitrate, cysteine, ascorbic acid, dicamba, glutamine into the Aa medium at the concentration of 4,40, 100, 2, and 5 mg L^-1, respectively. By using the Adi medium and the scraping technique, the regeneration frequencies of the mature embryos of CB037, Lunxuan 987, Hart 6172, Yangmai 6, Bobwhite, Zheng 9023, Shi 4 185, and Jimai 20 became 85.6, 60,1, 46.0, 42.1,42.0, 34.0, 33.0, and 32.0%, respectively, which were about 5-8 times higher than that obtained from the conventional culture mediums and techniques. This novel regeneration system could be helpful in wheat transformation.展开更多
The purpose of this study was to characterize Ta14 S homoeologs and assess their functions in wheat seed development.The genomic and c DNA sequences of three Ta14 S homoeologous genes encoding 14-3-3 proteins were iso...The purpose of this study was to characterize Ta14 S homoeologs and assess their functions in wheat seed development.The genomic and c DNA sequences of three Ta14 S homoeologous genes encoding 14-3-3 proteins were isolated.Sequence analysis revealed that the three homoeologs consisted of five exons and four introns and were very highly conserved in the coding regions and in exon/intron structure,whereas the c DNA sequences were variable in the 5′ and 3′-UTR.The three genes,designated as Ta14S-2A,Ta14S-2B and Ta14S-2D,were located in homoeologous group 2 chromosomes.The polypeptide chains of the three Ta14 S genes were highly similar.These genes were most homologous to Hv14 A from barley.Real-time quantitative PCR indicated that the three Ta14 S genes were differentially expressed in different organs at different developmental stages and all exhibited greater expression in primary roots of 1-day-old germlings than in other tissues.Comparison of the expression patterns of the three homoeologous genes at different times after pollination also revealed that their expression was developmentally regulated.The transcription of Ta14S-2B was clearly higher during seed germination,whereas expressions of Ta14S-2A and Ta14S-2D were up-regulated at the beginning of seed imbibition(0–12 h),but declined thereafter.The results suggested that the three Ta14 S homoeologous genes have regulatory roles in seed development and germination.展开更多
Using isobaric tags for relative and absolute quantification （iTRAQ） and associated analytic technologies, we have cataloged and compared 7 069 unique wheat proteins expressed during four substages of the filling st...Using isobaric tags for relative and absolute quantification （iTRAQ） and associated analytic technologies, we have cataloged and compared 7 069 unique wheat proteins expressed during four substages of the filling stage. Among them, 859 are differentially expressed, showing at least a 2-fold difference in concentration across substages. Differentially expressed proteins （DEPs） includind high-molecular weight giutenin subunit （W5AIU1）, low-molecular weight glutenin subunit （QSW3V4）, gliadin/avenin-like seed protein （D2KFG9）, and avenin-like protein （W5DVL2）, all of which have previously been identified as important for nutritional quality and bread-making properties, and all of which were found to increase at the latter stages of development. We have applied statistical techniques to group the proteins into hierarchical clusters, and have consulted databases to infer functional and other relationships among the identified proteins.展开更多
In the present study, one unique low-molecular-weight glutenin subunit （LMW-GS） gene. LMWXY22-2 （GenBank no. FJ028810）, was isolated from wheat cultivar Xiaoyan 22 （Triticum aestivum L.） by a pair of genomic spe...In the present study, one unique low-molecular-weight glutenin subunit （LMW-GS） gene. LMWXY22-2 （GenBank no. FJ028810）, was isolated from wheat cultivar Xiaoyan 22 （Triticum aestivum L.） by a pair of genomic specific PCR primers for 1B chromosome. Sequence analysis revealed that LMWXY22-2 was composed of 1 364 bp nucleotides, including a 317 bp promotion region and a 1 047 bp coding region which could be translated into a mature protein of 349 amino acids. In spite of a few minor mutations, the sequence of 5＇ untranslated region （UTR）, the coding region, the deduced N- and Cterminus comparisons indicated that LMWXY22-2 belonged to the reported subunits of LMW-m type and type lII group 5, respectively. Inner gene markers for 1D chromosome together with the phylogenetic analysis revealed that this gene was classified into Glu-D3, which was not in agreement with the I B locus-specific primers for LMW genes completely.展开更多
In order to study the roles of members of HKT gene fanfily in wheat, TaHKT2 ; 2 was isolated by using homologous cloning strategy and screening genomic BAC library. TaHKT2; 2 genes were mapped on chromosomes 7A, 7B an...In order to study the roles of members of HKT gene fanfily in wheat, TaHKT2 ; 2 was isolated by using homologous cloning strategy and screening genomic BAC library. TaHKT2; 2 genes were mapped on chromosomes 7A, 7B and 7D, named as TaHKT2; 2-7A, TaHKT2; 2-7B, and TaHKT2; 2-7D, respectively. TaHKT2 ; 2 and TaHKT2 ; 1 had the same genetic structure, composed of three exons and two introns, and formed a cluster with TaHKT2 ; 1 on the phylogenetic tree of plant HKT transporters. The coding sequences of TaHKT2 ; 2-7A, TaHKT2 ; 2-7B, and TaHKT2 ; 2-7D were 1 602, 1 602 and 1 596 bp long, respectively, but TaHKT2 ;2-7D cDNA sequence was not isolated by RT-PCR in eight wheat varieties. The natural diversity of TaHKT2 ;2 genes was analyzed by cloning and sequencing from 12 wheat varieties. The results showed that TaHKT2;2-7A was found to be more diverse than TaHKT2; 2-7B and TaHKT2; 2-7D. Only a few bases changed in the alleles of TaHKT2 ; 2 genes in wheat. No amino-acid natural variation lay in the P-loops of deduced protein sequences of all alleles of TaHKT2 ; 2 in 12 wheat varieties. The identity of coding sequences was nmch higher than that of 5＇ flanking regions of TaHKT2 ; 2 genes. TaHKT2 ; 2 nfight be selected over the comse of wheat domestication and belonged to domestication gene.展开更多
采用节水栽培并减少氮肥用量是实现豫北冬小麦生产的高产、高效和环境友好发展的必然选择,探明限水减氮对冬小麦产量和植株各层次器官干物质运转的影响,可为该地区冬小麦节水栽培和合理施用氮肥提供科学依据。2009-2010和2010-2011年连...采用节水栽培并减少氮肥用量是实现豫北冬小麦生产的高产、高效和环境友好发展的必然选择,探明限水减氮对冬小麦产量和植株各层次器官干物质运转的影响,可为该地区冬小麦节水栽培和合理施用氮肥提供科学依据。2009-2010和2010-2011年连续2年在河南浚县钜桥进行小麦田间裂区试验,主区设置2个灌溉水平[拔节水(W1)和拔节水+开花水(W2)],副区设置5个氮肥水平[330 kg hm^-2(N4,豫北地区小麦生产中常规施氮量)、270 kg hm^-2(N3)、210 kg hm^-2(N2)、120 kg hm^-2(N1)、0 kg hm^-2(N0)],测定了籽粒产量和植株各层次器官干物质运转量、运转率和对籽粒贡献率。减量施氮与N4相比,各营养器官向籽粒运转的干物质量均有增加,其中,穗轴+颖壳的干物质运转量增加了323.2%,增幅远高于茎节的24.5%和叶片的4.6%,且穗轴+颖壳的干物质运转率和对籽粒贡献率增幅也远高于茎节和叶片。减量施氮处理的叶片干物质运转量的增加主要源于倒三叶和倒四叶,分别增加28.7%和201.1%,而茎节干物质运转量的增加主要源于除穗位节外的其他茎节,分别增加21.7%(倒二节)、71.8%(倒三节)、44.5%(倒四节)和31.1%(余节)。与W2相比,W1干物质运转量无显著差异,但干物质运转率略高(24.6%vs.23.8%),对籽粒贡献率较高(35.1%vs.30.0%),籽粒产量降低11.2%,水分供应量减少750 m^3 hm^-2。可见,减量施氮促进了营养器官,尤其是穗轴+颖壳和下层器官(倒三叶、倒四叶、倒三节、倒四节和余节)的干物质向籽粒的运转,提高了对籽粒贡献率,有利于提高籽粒产量。展开更多
利用42个优质小麦(Triticum aestivum)品种籽粒发芽指数对6个小麦穗发芽抗性相关基因[Tamyb10、TaDFR(Dihydroflavone reductase)、TaVp-1(Viviparous‑1)、TaSdr(Seed dormancy)、TaPM19-A1(Plasma membrane 19‑A1)、TaMFT(Mother of FT...利用42个优质小麦(Triticum aestivum)品种籽粒发芽指数对6个小麦穗发芽抗性相关基因[Tamyb10、TaDFR(Dihydroflavone reductase)、TaVp-1(Viviparous‑1)、TaSdr(Seed dormancy)、TaPM19-A1(Plasma membrane 19‑A1)、TaMFT(Mother of FT and TFL1)]的分子标记的有效性进行验证,以期为抗穗发芽优质小麦品种的筛选及选育奠定基础。结果表明,小麦穗发芽抗性相关基因Tamyb10、TaDFR、TaVp-1、TaSdr、TaPM19-A1、TaMFT在42个优质小麦品种中均检测到2种等位变异,优异等位基因所占比例存在明显差异,介于4.8%~78.6%,未发现TaVp-1的TaVp-1 Bb基因型。等位基因类型与籽粒发芽指数相关性分析表明,标记myb10D、MFT-3A和MFT-A2与优质小麦穗发芽抗性极显著相关,而标记DFR-B、Vp1B3、Sdr2A、Sdr2B和PM19-A1与优质小麦穗发芽抗性相关性不显著。STS标记myb10D可用于红粒优质小麦的穗发芽抗性筛选,CAPS标记MFT-3A和STS标记MFT-A2可用于白粒优质小麦的穗发芽抗性筛选,TaMFT可能在优质小麦抗穗发芽机制中发挥着重要作用。展开更多
基金funded by the National Natural Science Foundation of China (30971776)the National Transgenic Organism Research Program of China(2008ZX08010-004)
文摘Establishment of a highly efficient regeneration system for the mature embryo of wheat will provide a convenient tool for wheat tissue culture and transformation, thereby facilitating the transformation of foreign genes into wheat. By using the mature embryos derived from 20 different wheat lines including Shi 4185, Yumai 66, Lunxuan 987, CB037, Yangmai 6, Xinchun 9, Bobwhite, Han 6172, Zheng 9023, Jimai 20, Ningchun 4, and Jing 411, the effects of some factors including inoculation methods, initiating culture media, organic additives, antioxidants, and auxins on the regeneration from the explants were evaluated. The results indicated that the scraping embryo culture was better than the whole embryo culture, the Aa medium was better than the SD2 medium and dicamba was better than 2,4-D in increasing the regeneration frequency. An Adi medium was established in this study by adding silver nitrate, cysteine, ascorbic acid, dicamba, glutamine into the Aa medium at the concentration of 4,40, 100, 2, and 5 mg L^-1, respectively. By using the Adi medium and the scraping technique, the regeneration frequencies of the mature embryos of CB037, Lunxuan 987, Hart 6172, Yangmai 6, Bobwhite, Zheng 9023, Shi 4 185, and Jimai 20 became 85.6, 60,1, 46.0, 42.1,42.0, 34.0, 33.0, and 32.0%, respectively, which were about 5-8 times higher than that obtained from the conventional culture mediums and techniques. This novel regeneration system could be helpful in wheat transformation.
基金financially supported by the Key Transgenic Breeding Program of the Ministry of Agriculture of China(No.2014ZX0800205B-003)the National Natural Science Foundation of China(No.30771332)
文摘The purpose of this study was to characterize Ta14 S homoeologs and assess their functions in wheat seed development.The genomic and c DNA sequences of three Ta14 S homoeologous genes encoding 14-3-3 proteins were isolated.Sequence analysis revealed that the three homoeologs consisted of five exons and four introns and were very highly conserved in the coding regions and in exon/intron structure,whereas the c DNA sequences were variable in the 5′ and 3′-UTR.The three genes,designated as Ta14S-2A,Ta14S-2B and Ta14S-2D,were located in homoeologous group 2 chromosomes.The polypeptide chains of the three Ta14 S genes were highly similar.These genes were most homologous to Hv14 A from barley.Real-time quantitative PCR indicated that the three Ta14 S genes were differentially expressed in different organs at different developmental stages and all exhibited greater expression in primary roots of 1-day-old germlings than in other tissues.Comparison of the expression patterns of the three homoeologous genes at different times after pollination also revealed that their expression was developmentally regulated.The transcription of Ta14S-2B was clearly higher during seed germination,whereas expressions of Ta14S-2A and Ta14S-2D were up-regulated at the beginning of seed imbibition(0–12 h),but declined thereafter.The results suggested that the three Ta14 S homoeologous genes have regulatory roles in seed development and germination.
基金supported by the National High-Tech R&D Program of China (863 Program,2011AA100501)the China Agricultural Research System (CARS-3-2-47)
文摘Using isobaric tags for relative and absolute quantification （iTRAQ） and associated analytic technologies, we have cataloged and compared 7 069 unique wheat proteins expressed during four substages of the filling stage. Among them, 859 are differentially expressed, showing at least a 2-fold difference in concentration across substages. Differentially expressed proteins （DEPs） includind high-molecular weight giutenin subunit （W5AIU1）, low-molecular weight glutenin subunit （QSW3V4）, gliadin/avenin-like seed protein （D2KFG9）, and avenin-like protein （W5DVL2）, all of which have previously been identified as important for nutritional quality and bread-making properties, and all of which were found to increase at the latter stages of development. We have applied statistical techniques to group the proteins into hierarchical clusters, and have consulted databases to infer functional and other relationships among the identified proteins.
基金supported by grants from the Significant Special Found of "13115" S&T Innovation Project of Shaanxi Province,China(2007 ZDKG-01)"13115" Technology Innovation Engineering and Engineering Technology Research Center of Shaanxi Province,China(2008 ZDGC-02)the Special Capital for the Construction of Modern Agriculture Technical System of Shaanxi Province,China (NYCYTX-001)
文摘In the present study, one unique low-molecular-weight glutenin subunit （LMW-GS） gene. LMWXY22-2 （GenBank no. FJ028810）, was isolated from wheat cultivar Xiaoyan 22 （Triticum aestivum L.） by a pair of genomic specific PCR primers for 1B chromosome. Sequence analysis revealed that LMWXY22-2 was composed of 1 364 bp nucleotides, including a 317 bp promotion region and a 1 047 bp coding region which could be translated into a mature protein of 349 amino acids. In spite of a few minor mutations, the sequence of 5＇ untranslated region （UTR）, the coding region, the deduced N- and Cterminus comparisons indicated that LMWXY22-2 belonged to the reported subunits of LMW-m type and type lII group 5, respectively. Inner gene markers for 1D chromosome together with the phylogenetic analysis revealed that this gene was classified into Glu-D3, which was not in agreement with the I B locus-specific primers for LMW genes completely.
基金Supported by National Key Research and Development Program of China(2016YFD0101802)Modern Agricultural Science and Technology Innovation Engineering Project of Hebei Province(2017038997)National Natural Science Foundation of Hebei Province(C2014106075)
文摘In order to study the roles of members of HKT gene fanfily in wheat, TaHKT2 ; 2 was isolated by using homologous cloning strategy and screening genomic BAC library. TaHKT2; 2 genes were mapped on chromosomes 7A, 7B and 7D, named as TaHKT2; 2-7A, TaHKT2; 2-7B, and TaHKT2; 2-7D, respectively. TaHKT2 ; 2 and TaHKT2 ; 1 had the same genetic structure, composed of three exons and two introns, and formed a cluster with TaHKT2 ; 1 on the phylogenetic tree of plant HKT transporters. The coding sequences of TaHKT2 ; 2-7A, TaHKT2 ; 2-7B, and TaHKT2 ; 2-7D were 1 602, 1 602 and 1 596 bp long, respectively, but TaHKT2 ;2-7D cDNA sequence was not isolated by RT-PCR in eight wheat varieties. The natural diversity of TaHKT2 ;2 genes was analyzed by cloning and sequencing from 12 wheat varieties. The results showed that TaHKT2;2-7A was found to be more diverse than TaHKT2; 2-7B and TaHKT2; 2-7D. Only a few bases changed in the alleles of TaHKT2 ; 2 genes in wheat. No amino-acid natural variation lay in the P-loops of deduced protein sequences of all alleles of TaHKT2 ; 2 in 12 wheat varieties. The identity of coding sequences was nmch higher than that of 5＇ flanking regions of TaHKT2 ; 2 genes. TaHKT2 ; 2 nfight be selected over the comse of wheat domestication and belonged to domestication gene.
文摘采用节水栽培并减少氮肥用量是实现豫北冬小麦生产的高产、高效和环境友好发展的必然选择,探明限水减氮对冬小麦产量和植株各层次器官干物质运转的影响,可为该地区冬小麦节水栽培和合理施用氮肥提供科学依据。2009-2010和2010-2011年连续2年在河南浚县钜桥进行小麦田间裂区试验,主区设置2个灌溉水平[拔节水(W1)和拔节水+开花水(W2)],副区设置5个氮肥水平[330 kg hm^-2(N4,豫北地区小麦生产中常规施氮量)、270 kg hm^-2(N3)、210 kg hm^-2(N2)、120 kg hm^-2(N1)、0 kg hm^-2(N0)],测定了籽粒产量和植株各层次器官干物质运转量、运转率和对籽粒贡献率。减量施氮与N4相比,各营养器官向籽粒运转的干物质量均有增加,其中,穗轴+颖壳的干物质运转量增加了323.2%,增幅远高于茎节的24.5%和叶片的4.6%,且穗轴+颖壳的干物质运转率和对籽粒贡献率增幅也远高于茎节和叶片。减量施氮处理的叶片干物质运转量的增加主要源于倒三叶和倒四叶,分别增加28.7%和201.1%,而茎节干物质运转量的增加主要源于除穗位节外的其他茎节,分别增加21.7%(倒二节)、71.8%(倒三节)、44.5%(倒四节)和31.1%(余节)。与W2相比,W1干物质运转量无显著差异,但干物质运转率略高(24.6%vs.23.8%),对籽粒贡献率较高(35.1%vs.30.0%),籽粒产量降低11.2%,水分供应量减少750 m^3 hm^-2。可见,减量施氮促进了营养器官,尤其是穗轴+颖壳和下层器官(倒三叶、倒四叶、倒三节、倒四节和余节)的干物质向籽粒的运转,提高了对籽粒贡献率,有利于提高籽粒产量。
文摘利用42个优质小麦(Triticum aestivum)品种籽粒发芽指数对6个小麦穗发芽抗性相关基因[Tamyb10、TaDFR(Dihydroflavone reductase)、TaVp-1(Viviparous‑1)、TaSdr(Seed dormancy)、TaPM19-A1(Plasma membrane 19‑A1)、TaMFT(Mother of FT and TFL1)]的分子标记的有效性进行验证,以期为抗穗发芽优质小麦品种的筛选及选育奠定基础。结果表明,小麦穗发芽抗性相关基因Tamyb10、TaDFR、TaVp-1、TaSdr、TaPM19-A1、TaMFT在42个优质小麦品种中均检测到2种等位变异,优异等位基因所占比例存在明显差异,介于4.8%~78.6%,未发现TaVp-1的TaVp-1 Bb基因型。等位基因类型与籽粒发芽指数相关性分析表明,标记myb10D、MFT-3A和MFT-A2与优质小麦穗发芽抗性极显著相关,而标记DFR-B、Vp1B3、Sdr2A、Sdr2B和PM19-A1与优质小麦穗发芽抗性相关性不显著。STS标记myb10D可用于红粒优质小麦的穗发芽抗性筛选,CAPS标记MFT-3A和STS标记MFT-A2可用于白粒优质小麦的穗发芽抗性筛选,TaMFT可能在优质小麦抗穗发芽机制中发挥着重要作用。
