Systemic virus-induced gene silencing allows functional characterization of maize genes during biotrophic interaction with Ustilago maydis
Systemic virus-induced gene silencing allows functional characterization of maize genes during biotrophic interaction with Ustilago maydis
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DOI：
10.1111/j.1469-8137.2010.03474.x
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发表时间：
2011-01-01
期刊：
影响因子：
9.4
通讯作者：
Doehlemann, Gunther
中科院分区：
文献类型：
--
作者：
van der Linde, Karina;Kastner, Christine;Doehlemann, Gunther
玉米（Zea mays）植株受玉米黑穗病菌（Ustilago maydis）侵染后，茎、叶和花序上形成较大的肿瘤。在这种生物营养相互作用中，植物防御反应受到病原体的积极抑制，先前对受感染玉米植株的转录组分析显示，在疾病进展过程中，寄主基因表达发生了大规模的阶段性变化。为了鉴定与玉米叶枯病相互作用有关的玉米基因，我们采用了一种基于玉米雀麦花叶病毒（BMV）的病毒诱导基因沉默（VIGS）系统。建立了bmv预侵染玉米植株的条件。这种设置可以使用基于实时荧光定量PCR （qRT-PCR）的读数来定量VIGS及其对美狄氏菌感染的影响。在原理验证实验中，一种美国麦冬菌诱导的萜烯合成酶被证明对疾病的发展具有负性调节作用，而一种参与细胞死亡抑制的蛋白质是美国麦冬菌完全毒力所必需的。结果表明，该系统是一种快速鉴定玉米基因的通用工具，可以确定玉米与玉米叶螟的亲和性。
P>Infection of maize (Zea mays) plants with the corn smut fungus Ustilago maydis leads to the formation of large tumors on the stem, leaves and inflorescences. In this biotrophic interaction, plant defense responses are actively suppressed by the pathogen, and previous transcriptome analyses of infected maize plants showed massive and stage-specific changes in host gene expression during disease progression.To identify maize genes that are functionally involved in the interaction with U. maydis, we adapted a virus-induced gene silencing (VIGS) system based on the brome mosaic virus (BMV) for maize. Conditions were established that allowed successful U. maydis infection of BMV-preinfected maize plants. This set-up enabled quantification of VIGS and its impact on U. maydis infection using a quantitative real-time PCR (qRT-PCR)-based readout.In proof-of-principle experiments, an U. maydis-induced terpene synthase was shown to negatively regulate disease development while a protein involved in cell death inhibition was required for full virulence of U. maydis.The results suggest that this system is a versatile tool for the rapid identification of maize genes that determine compatibility with U. maydis.