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多个外壳蛋白突变使转基因番茄中的马铃薯 rx 抗性基因无法识别 Pepino mosaic potexvirus(PepMV)。

Multiple coat protein mutations abolish recognition of Pepino mosaic potexvirus (PepMV) by the potato rx resistance gene in transgenic tomatoes.

机构信息

Equipe de Virologie, UMR GD2P, IBVM, INRA and Université Victor Ségalen Bordeaux2, BP81, Villenave d'Ornon Cedex, France.

出版信息

Mol Plant Microbe Interact. 2010 Apr;23(4):376-83. doi: 10.1094/MPMI-23-4-0376.

Abstract

Despite the fact that Pepino mosaic virus (PepMV) and Potato virus X (PVX) share less than 40% identity in their coat proteins (CP), the known PVX elicitor of Rx, transgenic tomato (cv. Microtom) plants expressing a functional potato Rx resistance gene showed resistance toward PepMV. However, in a low percentage of plants, PepMV accumulation was observed and back inoculation experiments demonstrated that these plants contained resistance-breaking PepMV variants. Sequencing of the CP gene of these variants showed the accumulation of mutations in the amino acid 41 to 125 region the CP, whereas no mutations were observed in the nonevolved isolates. Agroinfiltration-mediated transient expression of the mutant CP demonstrated that they had a greatly attenuated or abolished ability to induce a hypersensitive reaction in Rx-expressing Nicotiana benthamiana leaves. The transient expression of truncated forms of the PepMV CP allowed the identification of a minimal elicitor domain (amino acids 30 to 136). These results demonstrate that the Rx-based sensing system is able to recognize the PepMV CP but, contrary to the situation with PVX, for which only two closely spaced resistance-breaking mutations are known, many mutations over a significant stretch of the PepMV CP allow escape from recognition by Rx.

摘要

尽管 Pepino mosaic 病毒(PepMV)和马铃薯 X 病毒(PVX)的外壳蛋白(CP)仅有不到 40%的同源性,但已知的 PVX 诱导物 Rx,表达功能性马铃薯 Rx 抗性基因的转基因番茄(cv. Microtom)植株对 PepMV 表现出抗性。然而,在一小部分植株中观察到 PepMV 的积累,回接实验表明这些植株含有抗性丧失的 PepMV 变体。对这些变体的 CP 基因进行测序表明,CP 的氨基酸 41 到 125 区域积累了突变,而未进化的分离株则没有观察到突变。通过农杆菌介导的瞬时表达突变 CP 表明,它们诱导 Rx 表达的烟草原生质体叶片产生过敏反应的能力大大减弱或丧失。PepMV CP 的截断形式的瞬时表达允许鉴定出一个最小的诱导子结构域(氨基酸 30 到 136)。这些结果表明,基于 Rx 的感应系统能够识别 PepMV CP,但与 PVX 不同,对于 PVX,仅已知两个紧密间隔的抗性丧失突变,PepMV CP 上的许多突变允许逃避 Rx 的识别。

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