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
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 .
