Haynaldia villosa(2n=14,VV),a wild grass of the subtribe Triticeae,serves as potential gene resources for wheat genetic improvement.In this study,the proteome characterization during grain development of Yangmai 5 and...Haynaldia villosa(2n=14,VV),a wild grass of the subtribe Triticeae,serves as potential gene resources for wheat genetic improvement.In this study,the proteome characterization during grain development of Yangmai 5 and Yangmai 5-H.villosa 6VS/6AL translocation line was investigated by a comparative proteomic approach.Two-dimensional electrophoresis identified 81 differentially accumulated proteins(DAPs)during five grain developmental stages in wheat-H.villosa translocation line.These proteins were mainly involved in stress defense,storage protein,energy metabolism,protein metabolism and folding,carbon metabolism,nitrogen metabolism,and starch metabolism.In particular,6VS/6AL translocation led to significant upregulation of 36 DAPs and specific expression of 11 DAPs such as chitinase,thaumatin-like proteins,glutathione transferase,α-amylase inhibitor,heat shock proteins,and betaine aldehyde dehydrogenase.These proteins mainly involved in biotic and abiotic stress responses.Further analysis found that the upstream 1500 promoter regions of these stress-responsive DAP genes contained multiple high-frequency cis-acting elements related to stress defense such as abscisic acid response element ABRE,methyl jasmonate(MeJA)-response element TGACG-motif and CGTCA-motif involved in oxidative stress and antioxidant response element(ARE).RNA-seq and RT-qPCR analyses revealed the high expression of these stress-defensive DAP genes in the developing grains,particularly at the early-middle grain filling stages.Our results demonstrated that 6VS chromosome of H.villosa contains abundant stress-defensive proteins that have potential values for wheat genetic improvement.展开更多
Reverse_transcription Polymerase Chain Reaction (RT_PCR) was performed using cDNAs as templates from wheat_ Haynaldia villosa 6VS/6AL translocation line and 'Yangmai 5' induced with fungus Erysiphe gramin...Reverse_transcription Polymerase Chain Reaction (RT_PCR) was performed using cDNAs as templates from wheat_ Haynaldia villosa 6VS/6AL translocation line and 'Yangmai 5' induced with fungus Erysiphe graminis , and degenerate primers designed based on the conserved amino acid sequences of known plant disease_resistance genes. The cDNA sequences encoding cyclophilin_like and H +_ATPase_like genes were first isolated and characterized in wheat. The putative amino acid sequences of the two clones showed that they were highly homologous to those of cyclophilin proteins and H +_ATPases isolated from other plants. Thus they were designated as Ta_Cyp and Ta_MAH . The obvious expression differences could be observed between wheat_ H. villosa 6VS/6AL translocation line and susceptible wheat cultivar 'Yangmai 5', implying that the two genes may be related with the resistance of wheat_ H. villosa 6VS/6AL translocation line to disease. Southern blot indicated that the wheat genome contained 2-3 copies of Ta_Cyp gene and one copy of the Ta_MAH gene. Chinese Spring nulli_tetrasomic line analysis located the Ta_Cyp homologous genes on wheat chromosome 6A, 6B and 6D. Southern blot using Ta_Cyp clone as a probe showed that the polymorphic bands existed among the H. villosa , amphiploid of Triticum durum _ H. villosa , wheat_ H. villosa 6VS/6AL translocation line and 'Yangmai 5', suggesting that Ta_Cyp homologies exist in wheat genome as well as on the short arm of chromosome 6V in H. villosa .展开更多
To Investigate the mechanism of resistance to wheat （Triticum aestivum L.） powdery mildew, suppression subtractlve hybridization was conducted between an isogenic resistant line carrying Pm21 and its recurrent paren...To Investigate the mechanism of resistance to wheat （Triticum aestivum L.） powdery mildew, suppression subtractlve hybridization was conducted between an isogenic resistant line carrying Pm21 and its recurrent parent Yangmal 5 to Isolate the resistance relative genes. A cDNA fragment specifically expressed in the resistant line was obtained and its full length was cloned by in silico cloning and RT-PCR. This gene encoded a deduced protein of 219 amino acids with a leucine-rich repeat （LRR） motif, often found In plant resistance genes, and was designated as Ta-LRR2. Ta-LRR2 had an increased expression level in the resistant line after Inoculation with Erysiphe graminis DC. f. sp. tritici Marchal. PCR analysis with different cytogenetlc stocks suggested that Ta-LRR2 was specifically associated with chromosome arms 6VS and 6AS. Linkage analysis further showed that Ta-LRR2 could be used as a resistance gene analog polymorphism marker of Pm21 for marker-assisted selection in germplasm enhancement and breeding practice. Moreover, how to Isolate Pm21 based on the Information obtained for Ta-LRR2 is discussed.展开更多
基金This research was financially supported by the National Key R&D Program of China(2016YFD0100502)the National Natural Science Foundation of China(31771773)。
文摘Haynaldia villosa(2n=14,VV),a wild grass of the subtribe Triticeae,serves as potential gene resources for wheat genetic improvement.In this study,the proteome characterization during grain development of Yangmai 5 and Yangmai 5-H.villosa 6VS/6AL translocation line was investigated by a comparative proteomic approach.Two-dimensional electrophoresis identified 81 differentially accumulated proteins(DAPs)during five grain developmental stages in wheat-H.villosa translocation line.These proteins were mainly involved in stress defense,storage protein,energy metabolism,protein metabolism and folding,carbon metabolism,nitrogen metabolism,and starch metabolism.In particular,6VS/6AL translocation led to significant upregulation of 36 DAPs and specific expression of 11 DAPs such as chitinase,thaumatin-like proteins,glutathione transferase,α-amylase inhibitor,heat shock proteins,and betaine aldehyde dehydrogenase.These proteins mainly involved in biotic and abiotic stress responses.Further analysis found that the upstream 1500 promoter regions of these stress-responsive DAP genes contained multiple high-frequency cis-acting elements related to stress defense such as abscisic acid response element ABRE,methyl jasmonate(MeJA)-response element TGACG-motif and CGTCA-motif involved in oxidative stress and antioxidant response element(ARE).RNA-seq and RT-qPCR analyses revealed the high expression of these stress-defensive DAP genes in the developing grains,particularly at the early-middle grain filling stages.Our results demonstrated that 6VS chromosome of H.villosa contains abundant stress-defensive proteins that have potential values for wheat genetic improvement.
文摘Reverse_transcription Polymerase Chain Reaction (RT_PCR) was performed using cDNAs as templates from wheat_ Haynaldia villosa 6VS/6AL translocation line and 'Yangmai 5' induced with fungus Erysiphe graminis , and degenerate primers designed based on the conserved amino acid sequences of known plant disease_resistance genes. The cDNA sequences encoding cyclophilin_like and H +_ATPase_like genes were first isolated and characterized in wheat. The putative amino acid sequences of the two clones showed that they were highly homologous to those of cyclophilin proteins and H +_ATPases isolated from other plants. Thus they were designated as Ta_Cyp and Ta_MAH . The obvious expression differences could be observed between wheat_ H. villosa 6VS/6AL translocation line and susceptible wheat cultivar 'Yangmai 5', implying that the two genes may be related with the resistance of wheat_ H. villosa 6VS/6AL translocation line to disease. Southern blot indicated that the wheat genome contained 2-3 copies of Ta_Cyp gene and one copy of the Ta_MAH gene. Chinese Spring nulli_tetrasomic line analysis located the Ta_Cyp homologous genes on wheat chromosome 6A, 6B and 6D. Southern blot using Ta_Cyp clone as a probe showed that the polymorphic bands existed among the H. villosa , amphiploid of Triticum durum _ H. villosa , wheat_ H. villosa 6VS/6AL translocation line and 'Yangmai 5', suggesting that Ta_Cyp homologies exist in wheat genome as well as on the short arm of chromosome 6V in H. villosa .
基金Supported by the Hi-Tech Research and Development(863) Program of China(2001 AA222152,2003AA207100,and 2004AA222140)the National Natural Science Foundation of China and the Program for Changjiang Scholars and Innovative Research Team in University
文摘To Investigate the mechanism of resistance to wheat （Triticum aestivum L.） powdery mildew, suppression subtractlve hybridization was conducted between an isogenic resistant line carrying Pm21 and its recurrent parent Yangmal 5 to Isolate the resistance relative genes. A cDNA fragment specifically expressed in the resistant line was obtained and its full length was cloned by in silico cloning and RT-PCR. This gene encoded a deduced protein of 219 amino acids with a leucine-rich repeat （LRR） motif, often found In plant resistance genes, and was designated as Ta-LRR2. Ta-LRR2 had an increased expression level in the resistant line after Inoculation with Erysiphe graminis DC. f. sp. tritici Marchal. PCR analysis with different cytogenetlc stocks suggested that Ta-LRR2 was specifically associated with chromosome arms 6VS and 6AS. Linkage analysis further showed that Ta-LRR2 could be used as a resistance gene analog polymorphism marker of Pm21 for marker-assisted selection in germplasm enhancement and breeding practice. Moreover, how to Isolate Pm21 based on the Information obtained for Ta-LRR2 is discussed.
