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    张非亚, 杜运鹏, 刘瑞峰, 袁晓娜, 张冬梅, 贾桂霞. 月季NBS-LRR-RGAs的分离与鉴定[J]. 北京林业大学学报, 2016, 38(11): 89-96. DOI: 10.13332/j.1000-1522.20160092
    引用本文: 张非亚, 杜运鹏, 刘瑞峰, 袁晓娜, 张冬梅, 贾桂霞. 月季NBS-LRR-RGAs的分离与鉴定[J]. 北京林业大学学报, 2016, 38(11): 89-96. DOI: 10.13332/j.1000-1522.20160092
    ZHANG Fei-ya, DU Yun-peng, LIU Rui-feng, YUAN Xiao-na, ZHANG Dong-mei, JIA Gui-xia. Isolation and characterization of NBS-LRR resistance gene candidates in rose.[J]. Journal of Beijing Forestry University, 2016, 38(11): 89-96. DOI: 10.13332/j.1000-1522.20160092
    Citation: ZHANG Fei-ya, DU Yun-peng, LIU Rui-feng, YUAN Xiao-na, ZHANG Dong-mei, JIA Gui-xia. Isolation and characterization of NBS-LRR resistance gene candidates in rose.[J]. Journal of Beijing Forestry University, 2016, 38(11): 89-96. DOI: 10.13332/j.1000-1522.20160092

    月季NBS-LRR-RGAs的分离与鉴定

    Isolation and characterization of NBS-LRR resistance gene candidates in rose.

    • 摘要: RGAs已经在很多植物中得到分离与克隆,它往往是R基因或者是R基因的一部分。为了分离与鉴定与月季抗黑斑病相关的RGAs,在转录组数据基础上,设计简并引物6对,从现代月季、疏花蔷薇、‘白玫瑰’上分离克隆了 NBS-LRR-RGAs,挑取750个克隆进行序列分析。从系统发育树可以看出:所克隆的RGAs与已克隆的其他植物的R基因具有较近的亲缘关系,分为non-TIR-NBS-RGAs和TIR-NBS-RGAs 2个大类,10个进化枝,其中所克隆得到的non-TIR-NBS-RGAs的比例远大于TIR-NBS-RGAs的比例;氨基酸序列比对发现所克隆的RGAs均含有完整的NBS结构域P-Loop、kinase-2、kinase-3a和GLPL;通过进化树对不同家族的6个 RGAs 的表达分析可看出,它们都不同程度、不同时间地参与了抗病反应。

       

      Abstract: RGAs have been isolated and characterized from many different plant species, and they are often located in genomic regions containing R genes. For the isolation and identification of RGAs, which were related to rose resistance to black pot, six pairs of degenerate primers were designed based on the transcriptome sequencing, and the NBS-LRR-RGAs were isolated and cloned from modern rose, Rosa laxa and Rosa rugosa ‘White’. 750 clones were selected and sequenced. Phylogenetic studies revealed that the sequenced rose RGAs clones showed a close relationship with R genes and these sequences were classified into ten clades, with non-TIR-NBS-RGAs and TIR-NBS-RGAs subclasses. The proportion of cloned non-TIR-NBS-RGAs was far greater than that of TIR-NBS-RGAs. The deduced amino acid sequences of RGAs all contained the whole conserved domains: P-Loop,kinase-2,kinase-3a and GLPL. The analysis of quantitative real-time PCR shows that the six RGAs from different clade participate in the reaction in different transcript levels and different time.

       

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