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来自番茄(Lycopersicon esculentum)的持久抗番茄花叶病毒基因Tm-2(2)的克隆与特性分析

Cloning and characterization of the durable tomato mosaic virus resistance gene Tm-2(2) from Lycopersicon esculentum.

作者信息

Lanfermeijer Frank C, Dijkhuis Jos, Sturre Marcel J G, de Haan Peter, Hille Jacques

机构信息

Department Molecular Biology of Plants, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands.

出版信息

Plant Mol Biol. 2003 Jul;52(5):1037-49. doi: 10.1023/a:1025434519282.

Abstract

In tomato, infections by tomato mosaic virus are controlled by durable Tm-2(2) resistance. In order to gain insight into the processes underlying disease resistance and its durability, we cloned and analysed the Tm-2(2) resistance gene and the susceptible allele, tm-2. The Tm-2(20 gene was isolated by transposon tagging using a screen in which plants with a destroyed Tm-2(2) gene survive. The Tm-2(2) locus consists of a single gene that encodes an 861 amino acid polypeptide, which belongs to the CC-NBS-LRR class of resistance proteins. The putative tm-2 allele was cloned from susceptible tomato lines via PCR with primers based on the Tm-2(2) sequence. Interestingly, the tm-2 gene has an open reading frame that is comparable to the Tm-2(2) allele. Between the tm-2 and the Tm-2(2) polypeptide 38 amino acid differences are present of which 26 are located in the second half of the LRR-domain. Susceptible tomato plants, which were transformed with the Tm-2(2) gene, displayed resistance against ToMV infection. In addition, virus specificity, displayed by the Tm-2(2) resistance was conserved in these transgenic lines. To explain the durability of this resistance, it is proposed that the Tm-2(2)-encoded resistance is aimed at the Achilles' heel of the virus.

摘要

在番茄中,番茄花叶病毒的感染由持久的Tm-2(2)抗性控制。为了深入了解抗病性及其持久性的潜在过程,我们克隆并分析了Tm-2(2)抗性基因和易感等位基因tm-2。通过转座子标签法,利用一种筛选方法分离出Tm-2(2)基因,在该筛选中,具有被破坏的Tm-2(2)基因的植物能够存活。Tm-2(2)位点由一个单一基因组成,该基因编码一个861个氨基酸的多肽,属于抗性蛋白的CC-NBS-LRR类。假定的tm-2等位基因是通过基于Tm-2(2)序列的引物进行PCR从易感番茄品系中克隆出来的。有趣的是,tm-2基因有一个与Tm-2(2)等位基因相当的开放阅读框。在tm-2和Tm-2(2)多肽之间存在38个氨基酸差异,其中26个位于LRR结构域的后半部分。用Tm-2(2)基因转化的易感番茄植株对番茄花叶病毒感染表现出抗性。此外,这些转基因品系中保留了Tm-2(2)抗性所表现出的病毒特异性。为了解释这种抗性的持久性,有人提出Tm-2(2)编码的抗性针对的是病毒的致命弱点。

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