Development and use of a reverse transcription-PCR assay to study expression of <i>Tri5</i> by <i>Fusarium</i> species in vitro and in planta

作     者：Doohan, FM Weston, G Rezanoor, HN Parry, DW Nicholson, P 

作者机构：John Innes Ctr Plant Sci Res Cereals Dept Norwich NR4 7UH Norfolk England Hort Res Int Maidstone ME19 6BJ Kent England 

出 版 物：《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 (应用与环境微生物学)

年 卷 期：1999年第65卷第9期

页      面：3850-3854页

核心收录：

学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 07[理学] 0836[工学-生物工程] 071005[理学-微生物学] 10[医学] 

主　　题：抗真菌药/药理学 碳碳裂合酶类/生物合成 碳碳裂合酶类/遗传学 DNA 真菌/分析 镰刀菌属/药物作用 镰刀菌属/酶学 镰刀菌属/遗传学 镰刀菌属/分离和提纯 基因表达 植物疾病/微生物学 逆转录聚合酶链反应/方法 三唑类/药理学 单端孢霉烯类/代谢 小麦属/微生物学 

摘      要：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 deoxunivalenol 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.