用于研究镰刀菌属物种在体外和植物体内Tri5基因表达的逆转录-聚合酶链反应分析方法的开发与应用。
Development and use of a reverse transcription-PCR assay to study expression of Tri5 by Fusarium species in vitro and in planta.
作者信息
Doohan F M, Weston G, Rezanoor H N, Parry D W, Nicholson P
机构信息
Cereals Department, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, United Kingdom.
出版信息
Appl Environ Microbiol. 1999 Sep;65(9):3850-4. doi: 10.1128/AEM.65.9.3850-3854.1999.
Abstract
The Tri5 gene encodes trichodiene synthase, which catalyzes the first reaction in the trichothecene biosynthetic pathway. In vitro, a direct relationship was observed between Tri5 expression and the increase in deoxynivalenol production over time. We developed a reverse transcription (RT)-PCR assay to quantify Tri5 gene expression in trichothecene-producing strains of Fusarium species. We observed an increase in Tri5 expression following treatment of Fusarium culmorum with fungicides, and we also observed an inverse relationship between Tri5 expression and biomass, as measured by beta-D-glucuronidase activity, during colonization of wheat (cv. Avalon) seedlings by F. culmorum. RT-PCR analysis also showed that for ears of wheat cv. Avalon inoculated with F. culmorum, there were different levels of Tri5 expression in grain and chaff at later growth stages. We used the Tri5-specific primers to develop a PCR assay to detect trichothecene-producing Fusarium species in infected plant material.
摘要
Tri5基因编码单端孢霉烯合酶，该酶催化单端孢霉烯生物合成途径中的第一步反应。在体外实验中，观察到Tri5的表达与脱氧雪腐镰刀菌烯醇产量随时间增加之间存在直接关系。我们开发了一种逆转录（RT）-PCR检测方法，用于定量镰刀菌属产单端孢霉烯菌株中Tri5基因的表达。在用杀菌剂处理禾谷镰刀菌后，我们观察到Tri5的表达增加，并且在禾谷镰刀菌定殖小麦（品种Avalon）幼苗期间，通过β-D-葡萄糖醛酸酶活性测量，我们还观察到Tri5表达与生物量之间呈负相关。RT-PCR分析还表明，对于接种了禾谷镰刀菌的小麦品种Avalon的麦穗，在后期生长阶段，籽粒和颖壳中的Tri5表达水平不同。我们使用Tri5特异性引物开发了一种PCR检测方法，以检测受感染植物材料中产单端孢霉烯的镰刀菌属物种。
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