Cloning, structural features, and expression analysis of resistance gene analogs in Tobacco
Cloning, structural features, and expression analysis of resistance gene analogs in Tobacco
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DOI：
10.1007/s11033-009-9749-2
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发表时间：
2010-01-01
影响因子：
2.8
通讯作者：
Lu, Xiuping
中科院分区：
文献类型：
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作者：
Gao, Yulong;Xu, Zhaoli;Lu, Xiuping
利用基于保守核苷酸结合位点(NBS)和蛋白激酶结构域(PKD)的简并引物，从烟草品种Repanda中分离到100个抗性基因类似物(RGA)。在GenBank数据库中搜索BLASTx，发现27个属于NBS类，73个属于蛋白激酶(PK)类。对推导的蛋白质序列进行聚类分析和多序列比对表明，NBS类的RGA可分为两类：Toll/白细胞介素型受体(TIR)和非TIR型。这两种类型都有6个保守基序(P-loop、RNBS-A、Kinase-2、RNBS-B、RNBS-C、GLPL)。根据它们的序列相似性，将PK类的烟草RGA分为8个亚类。我们检测了它们在烟草花叶病毒(TMV)或烟草黑胫病(Phytophthora parasitica var.)侵染后的表达。尼古丁)。4个PK类RGA被TMV和1个PK类RGA显著上调，3个NBS类RGA被寄生P.parasitica var上调。尼古丁。PK类的两个RGA被寄生的P.parasitica var诱导表达。尼古丁。侵染烟草花叶病毒或寄生疫霉变种。尼古丁促进PK类RGA基因NtRGA2的表达。本研究表明烟草基因组中含有丰富的RGA，对病原菌诱导的烟草RGA的鉴定将为烟草抗病相关基因的克隆提供有价值的信息。
Using degenerate primers based on the conserved nucleotide binding site (NBS) and protein kinase domain (PKD), 100 resistance gene analogs (RGAs) were isolated from tobacco variety Nicotiana repanda. BLASTx search against the GenBank database revealed that 27 belong to the NBS class and 73 belong to the protein kinase (PK) class. Cluster analysis and multiple sequence alignment of the deduced protein sequences indicate that RGAs of the NBS class can be divided into two groups: toll/interleukin receptor (TIR) and non-TIR types. Both types possess 6 conserved motifs (P-loop, RNBS-A, Kinase-2, RNBS-B, RNBS-C, GLPL). Based on their sequence similarity, the tobacco RGAs of the PK class were assigned to 8 subclasses. We examined their expression after infection with either Tobacco mosaic virus (TMV) or the tobacco black shank pathogen (Phytophthora parasitica var. nicotianae). The expression levels of 4 RGAs of the PK class were significantly elevated by TMV and 1 RGA of the PK class and 3 RGAs of the NBS class were up-regulated by P. parasitica var. nicotianae. The expression of two RGAs of the PK class was induced by P. parasitica var. nicotianae. Infection by either TMV or P. parasitica var. nicotianae enhanced the expression of NtRGA2, a RGA of the PK class. The present study shows that RGAs are abundant in the tobacco genome and the identification of tobacco RGAs induced by pathogens should provide valuable information for cloning related resistance genes in tobacco.