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伏马菌素B1介导的细胞死亡模拟及抗伏马菌素B1(fbr)拟南芥突变体的筛选

Simulation of fungal-mediated cell death by fumonisin B1 and selection of fumonisin B1-resistant (fbr) Arabidopsis mutants.

作者信息

Stone J M, Heard J E, Asai T, Ausubel F M

机构信息

Department of Molecular Biology, Massachusetts General Hospital, and Department of Genetics, Harvard Medical School, Boston, Massachusetts 02114, USA.

出版信息

Plant Cell. 2000 Oct;12(10):1811-22. doi: 10.1105/tpc.12.10.1811.

Abstract

Fumonisin B1 (FB1), a programmed cell death-eliciting toxin produced by the necrotrophic fungal plant pathogen Fusarium moniliforme, was used to simulate pathogen infection in Arabidopsis. Plants infiltrated with 10 microM FB1 and seedlings transferred to agar media containing 1 microM FB1 develop lesions reminiscent of the hypersensitive response, including generation of reactive oxygen intermediates, deposition of phenolic compounds and callose, accumulation of phytoalexin, and expression of pathogenesis-related (PR) genes. Arabidopsis FB1-resistant (fbr) mutants were selected directly by sowing seeds on agar containing 1 microM FB1, on which wild-type seedlings fail to develop. Two mutants chosen for further analyses, fbr1 and fbr2, had altered PR gene expression in response to FB1. fbr1 and fbr2 do not exhibit differential resistance to the avirulent bacterial pathogen Pseudomonas syringae pv maculicola (ES4326) expressing the avirulence gene avrRpt2 but do display enhanced resistance to a virulent isogenic strain that lacks the avirulence gene. Our results demonstrate the utility of FB1 for high-throughput isolation of Arabidopsis defense-related mutants and suggest that pathogen-elicited programmed cell death of host cells may be an important feature of compatible plant-pathogen interactions.

摘要

伏马菌素B1(FB1)是由坏死性真菌植物病原菌串珠镰刀菌产生的一种引发程序性细胞死亡的毒素,被用于模拟拟南芥中的病原体感染。用10微摩尔FB1浸润的植株以及转移至含有1微摩尔FB1的琼脂培养基上的幼苗会形成类似于过敏反应的损伤,包括活性氧中间体的产生、酚类化合物和胼胝质的沉积、植保素的积累以及病程相关(PR)基因的表达。通过将种子播种在含有1微摩尔FB1的琼脂上直接筛选出拟南芥FB1抗性(fbr)突变体,野生型幼苗在这种琼脂上无法生长。选择用于进一步分析的两个突变体fbr1和fbr2,其PR基因表达在对FB1的反应中发生了改变。fbr1和fbr2对表达无毒基因avrRpt2的无毒细菌病原体丁香假单胞菌番茄致病变种(ES4326)没有表现出差异抗性,但对缺乏无毒基因的同源致病菌株表现出增强的抗性。我们的结果证明了FB1在高通量分离拟南芥防御相关突变体中的实用性,并表明病原体引发的宿主细胞程序性死亡可能是植物与病原体亲和性相互作用的一个重要特征。

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