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中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (31): 21-26.doi: 10.11924/j.issn.1000-6850.2013-1414

所属专题： 生物技术； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

巴西橡胶树一个Mlo基因克隆及其序列特征分析

覃碧 王萌 薛松 胡中昀 郑服丛 张宇   

  • 收稿日期:2013-05-20 修回日期:2013-06-24 出版日期:2013-11-05 发布日期:2013-11-05
  • 基金资助:
    农业部热作专项“橡胶树病虫害监测防控技术研究”(12RZBC-11)；海南省研究生创新科研课题“基于物联网的橡胶树病虫害监测预警及决策建议平 台构建”(Hyb2012-4)；现代农业产业技术体系建设专项资金(CARS-34-GW8)

Cloning and Sequence Characterizing a Mlo Gene in Hevea brasiliensis Mull. Arg.

Qin Bi1, Wang Meng2, Xue Song3, Hu Zhongyun3, Zheng Fucong2, Zhang Yu2   

  • Received:2013-05-20 Revised:2013-06-24 Online:2013-11-05 Published:2013-11-05

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摘要： Mlo基因是植物抗病基因中的重要成员。巴西橡胶树Mlo基因的克隆和序列分析，将为进一步通过转基因技术培育抗病橡胶树新品种奠定基础。本研究以大麦Mlo基因(GenBank登录号: Z83834)为探针对巴西橡胶树的EST和转录组数据库进行同源搜索，并将获得的同源EST序列进行拼接，最后得到巴西橡胶树Mlo基因的cDNA序列。采用RT-PCR方法成功克隆了与预期大小一致的Mlo基因CDS片段1668 bp，编码555个氨基酸，分子量为63.14 kDa，等电点是8.85。对巴西橡胶树Mlo的编码蛋白结构域进行分析发现，该蛋白具有Mlo保守结构域，包含8次跨膜结构。通过对不同物种的Mlo同源蛋白进行聚类分析发现，巴西橡胶树Mlo蛋白与木薯Mlo蛋白同源性最高，达到92.1%，其次是蓖麻(87.6%)；在拟南芥基因组中，与AtMlo8的相似性最高，达到67%，而且AtMlo8也包含有8次跨膜结构，因此，将巴西橡胶树中克隆的Mlo基因命名为HbMlo8。通过对所克隆的HbMlo8基因分析可知，该基因属于Mlo基因家族成员，基因的克隆及序列分析为进一步验证巴西橡胶树HbMlo8的功能奠定了基础。

关键词: 实践, 实践

Abstract: Mlo is an important member of plant diseases resistance genes. Cloning and sequence analysis of Mlo gene in Hevea brasiliensis will lay the foundation for breeding for disease resistant varieties through the transgenic technology. In this study, the barley Mlo gene (GenBank accession number: Z83834) was served as a query to search in the EST and transcript databases of Hevea brasiliensis, and then the homologous EST were assembled. The CDS fragment of Hevea Mlo gene with the expected size (1668 bp) has been successfully cloned by RT-PCR. The Hevea Mlo encoded 555 amino acid residues, with a predicted molecular mass of 63.14 kDa and a 8.85 isolectric point. The structure analysis revealed that Hevea Mlo protein had a conserved Mlo domain and 8 transmembrane domains. Phylogenetic analysis showed that Hevea Mlo shared the highest similarity with the Mlo protein from Manihot esculenta and Ricinus communis, with 92.1% and 87.6% sequence identities. In the genome of Arabidopsis thaliana, Hevea Mlo shared the highest similarity with AtMlo8 with 67% identical residues, which also contained 8 transmembrane domains. Therefore, the Hevea Mlo cloned in this study was designed as HbMlo8. Cloning and sequence analysis of HbMlo8 gene suggested that it belonged to Mlo gene family, these results provide a basis for further verify its function in Hevea brasiliensis.

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