Isolation, characterization and phylogenetic analysis of the resistance gene analogues (RGAs) in banana (Musa spp.)
Isolation, characterization and phylogenetic analysis of the resistance gene analogues (RGAs) in banana (Musa spp.)
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DOI：
10.1016/j.plantsci.2007.02.019
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发表时间：
2007-06
期刊：
影响因子：
5.2
通讯作者：
X. Pei;Sheng-Hui Li;Ying Jiang;Yongqiang Zhang;Zhi-Xing Wang;S. Jia
中科院分区：
文献类型：
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作者：
X. Pei;Sheng-Hui Li;Ying Jiang;Yongqiang Zhang;Zhi-Xing Wang;S. Jia
根据香蕉抗病基因(R基因)核苷酸结合位点的保守区和富亮氨酸重复序列(NBS-LRR)，采用简并引物聚合酶链式反应(PCR)技术，分别从香蕉(Musa spp.)野生种工椒(AA)和新一叶椒(BB)，栽培种中山大椒(ABB)、粉椒(AAB)和威廉姆斯(AAA)中分离出R基因类似物。结果表明，在测序的208个克隆中，共有91个具有不间断开放阅读框架(ORF)的序列是典型的RGA，其中33个氨基酸序列的同源性低于97%。基于MEGA软件的系统发育分析，33个序列可以划分为12个不同的Musa RGA。它们都属于非TIR-NBS类型。用TIR-型特异性引物进行扩增，未检测到代表的TIR-NBS-R基因，说明在香蕉进化过程中，TIR-NBS-型R基因发生了选择性缺失。通过与其他植物中已知的R基因的比较和系统发育分析，揭示了它们之间的进化关系。尽管在香蕉中发现了高度多样性的RGA，但除了两个Musa RGA外，在所有供试香蕉中都检测到了12个Musa RGA：Musa RGA-I仅在中山大角(ABB)和拱角(AA)中存在和表达，而Musa RGA-K仅在中山大角中出现和表达，这表明野生和栽培香蕉之间总体上存在轻微的分化。据我们所知，到目前为止还没有关于香蕉中RGA的分离、鉴定和系统发育分析的报道。
According to the conservative regions of the nucleotide-binding site and the leucine-rich repeat (NBS-LRR) in resistance genes (R genes), the polymerase chain reaction (PCR) with degenerate primers was employed to isolate R gene analogues (RGAs) from five species of banana (Musa spp.), i.e. wild species of Gongjiao (AA) and Xinyiyejiao (BB), cultivated species of Zhongshandajiao (ABB), Fenjiao (AAB) and Williams (AAA), respectively. As a result, a total of 91 sequences with uninterrupted open reading frame (ORFs) were typical of RGAs out of 208 clones sequenced, of which 33 sequences with identity of deduced amino acid sequence below 97% were further analyzed. Based on the phylogenetic analysis by using MEGA software, the 33 sequences could be divided into 12 distinct Musa RGAs. All of them belong to the non-TIR-NBS type. No representatives of the TIR-type RGAs were detected by PCR amplification using TIR-type specific primers, suggesting the TIR-NBS-type of R genes were selectively lost in banana during evolution process. Comparison and phylogenetic analysis of the Musa RGAs with the known R genes from other plant species revealed their evolutionary relationship. Despite the high diversity of the RGAs found in banana, the 12 Musa RGAs were detected in all banana species tested, with the exception of two Musa RGAs: Musa RGA-I only presented and expressed in Zhongshandajiao (ABB) and Gongjiao (AA), while Musa RGA-K only appeared and expressed in Zhongshandajiao, suggesting there was minor differentiation between wild and cultivated forms in general. To the best of our knowledge, the isolation, characterization and phylogenetic analysis of RGAs in banana has not been reported in the literature to date.