为了筛选抗白粉病基因Pm21介导的抗病途径中的防卫基因,采用芯片杂交技术筛选携带或不携带Pm21基因的材料之间或抗病材料经白粉菌诱导与非诱导样品之间的差异表达基因,并采用RT-PCR方法分析了被筛选出的草酸盐氧化酶类似蛋白基因(Oxalat...为了筛选抗白粉病基因Pm21介导的抗病途径中的防卫基因,采用芯片杂交技术筛选携带或不携带Pm21基因的材料之间或抗病材料经白粉菌诱导与非诱导样品之间的差异表达基因,并采用RT-PCR方法分析了被筛选出的草酸盐氧化酶类似蛋白基因(Oxalate Oxidase Like Protein,OxOLP)的表达情况,进一步利用TAIL-PCR方法分离簇毛麦基因Hv-OxOLP中的全长序列。芯片杂交结果表明,探针Contig3156(一个推导的草酸盐氧化酶类似蛋白基因)在抗病簇毛麦中受白粉菌诱导的程度最大,在抗病易位系中该探针的杂交信号比在感病扬麦5号中的杂交信号强。半定量RT-PCR结果表明,在抗、感白粉病的材料中草酸盐氧化酶类似蛋白基因都受白粉菌诱导表达,但在抗病材料中达到表达峰值的时间早。用TAIL-PCR方法分离的Hv-OxOLP有一个内含子和两个外显子,ORF包含690个核苷酸,在启动子区包含两个W-box,在终止密码子后面还有一个polyA的加尾信号。作为一个受白粉菌诱导表达的基因,Hv-OxOLP在抗白粉病反应中可能起重要的作用。展开更多
Plant WRKY transcription factors are involved in various physiological processes, including biotic and abiotic stress responses, as well as developmental processes. In this study, the expression patterns of the WRKY68...Plant WRKY transcription factors are involved in various physiological processes, including biotic and abiotic stress responses, as well as developmental processes. In this study, the expression patterns of the WRKY68 protein during interactions between rice 4021 containing the bacterial blight resistance gene Xa21 and Xanthomonas oryzae pv. oryzae（Xoo） were investigated. A possible modified form of the WRKY68 protein appeared in the Xa21-mediated disease resistance response, and its expression levels were similar in compatible and incompatible responses, but differed significantly from that of the mock control treatment, suggesting that WRKY68 may be involved in the bacterial blight response in rice. To further understand WRKY68＇s roles in the resistance signaling pathway, WRKY68 recombinant protein was expressed in Escherichia coli and a microscale thermophoresis analysis was performed to investigate the interactions between WRKY68 and cis-elements in crucial pathogenesis-related（PR） genes. The results showed that the WRKY68 protein binds to W-boxes in the PR1 b promoter region, with an apparent dissociation constant of 25 nmol L–1, while the binding between WRKY68 and PR10 a was W-box independent. The results suggested that a possible modified form of the WRKY68 protein was induced during the interaction between rice and Xoo, which then regulated the activity of the downstream PR genes by binding with the W-boxes in the PR1 b gene＇s promoter region. Moreover, the constitutive transcription of the WRKY68 gene in dozens of rice tissues and the expression of the WRKY68 protein in leaves during all growth stages suggests that WRKY68 plays important roles in rice during normal growth processes.展开更多
基金Special Financial Grant from China Postdoctral Science Foundation(2014T70373)54-class General Financial Grant from China Postdoctral Science Foundation(2013M541422)+3 种基金Department of Science and Technology of Heilongjiang Province(LC2011C30)Harbin Science and Technology Bureau(2011RFLYN019)Young Foundation Major Project of HAAS(2012ZD031)Department of Human Resource of Heilongjiang Province(2012-134)
文摘为了筛选抗白粉病基因Pm21介导的抗病途径中的防卫基因,采用芯片杂交技术筛选携带或不携带Pm21基因的材料之间或抗病材料经白粉菌诱导与非诱导样品之间的差异表达基因,并采用RT-PCR方法分析了被筛选出的草酸盐氧化酶类似蛋白基因(Oxalate Oxidase Like Protein,OxOLP)的表达情况,进一步利用TAIL-PCR方法分离簇毛麦基因Hv-OxOLP中的全长序列。芯片杂交结果表明,探针Contig3156(一个推导的草酸盐氧化酶类似蛋白基因)在抗病簇毛麦中受白粉菌诱导的程度最大,在抗病易位系中该探针的杂交信号比在感病扬麦5号中的杂交信号强。半定量RT-PCR结果表明,在抗、感白粉病的材料中草酸盐氧化酶类似蛋白基因都受白粉菌诱导表达,但在抗病材料中达到表达峰值的时间早。用TAIL-PCR方法分离的Hv-OxOLP有一个内含子和两个外显子,ORF包含690个核苷酸,在启动子区包含两个W-box,在终止密码子后面还有一个polyA的加尾信号。作为一个受白粉菌诱导表达的基因,Hv-OxOLP在抗白粉病反应中可能起重要的作用。
基金supported by the Specialized Research Fund for the Doctoral Program of Higher Education,China(20131302110006)
文摘Plant WRKY transcription factors are involved in various physiological processes, including biotic and abiotic stress responses, as well as developmental processes. In this study, the expression patterns of the WRKY68 protein during interactions between rice 4021 containing the bacterial blight resistance gene Xa21 and Xanthomonas oryzae pv. oryzae（Xoo） were investigated. A possible modified form of the WRKY68 protein appeared in the Xa21-mediated disease resistance response, and its expression levels were similar in compatible and incompatible responses, but differed significantly from that of the mock control treatment, suggesting that WRKY68 may be involved in the bacterial blight response in rice. To further understand WRKY68＇s roles in the resistance signaling pathway, WRKY68 recombinant protein was expressed in Escherichia coli and a microscale thermophoresis analysis was performed to investigate the interactions between WRKY68 and cis-elements in crucial pathogenesis-related（PR） genes. The results showed that the WRKY68 protein binds to W-boxes in the PR1 b promoter region, with an apparent dissociation constant of 25 nmol L–1, while the binding between WRKY68 and PR10 a was W-box independent. The results suggested that a possible modified form of the WRKY68 protein was induced during the interaction between rice and Xoo, which then regulated the activity of the downstream PR genes by binding with the W-boxes in the PR1 b gene＇s promoter region. Moreover, the constitutive transcription of the WRKY68 gene in dozens of rice tissues and the expression of the WRKY68 protein in leaves during all growth stages suggests that WRKY68 plays important roles in rice during normal growth processes.
