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Identification and functional characterization of cryptic secondary metabolite gene clusters and comparative genomics in Fusarium fujikuroi

藤黑镰刀菌隐性次生代谢基因簇的鉴定和功能表征以及比较基因组学

基本信息

批准号：
175227329
负责人：
Dr. Ulrich Güldener
金额：
--
依托单位：
Deutsches Herzzentrum München (DHM)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2010
资助国家：
德国
起止时间：
2009-12-31 至 2016-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/175227329?language=en
关键词：
Identification functional characterization cryptic secondary

项目摘要

Species of the genus Fusarium are well known as serious plant pathogens and producers of several harmful mycotoxins, such as fumonisins, moniliformin, zearalenone, fusarin C, fusaric acid, bikaverin, equisetin, and beauvericin. With the complete genome sequence of the maize pathogen F. verticillioides at hand, and the intended genome sequencing of the biotechnologically important (gibberellin production) fungus F. fujikuroi, we will be able to identify the genes (clusters) for biosynthesis of two harmful polyketide mycotoxins: moniliformin and fusarin C. To date, none of the moniliformin biosynthetic genes is known from any Fusarium species, and only one fusarin C biosynthetic gene, the hybridpolyketide synthase/ non ribosomal peptidsynthase (PKS/NRPS)-encoding gene, has been identified in Fusarium moniliformis. In order to reduce the health risk of these mycotoxins in food, feed and biotechnologically produced gibberellin preparations, we are planning to identify the yet unknown genes and pathways for biosynthesis of fusarin C and moniliformin in F. verticillioides and F. fujikuroi. To identify the moniliformin-specific PKS gene, we will generate knock-out mutants for candidate PKS genes in a moniliformin-producing F. fujikuroi strain and identify a moniliformin-minus mutant. For fusarin C, we will delete more putative biosynthetic genes adjacent to the already known PKS/NRPS gene and identify the intermediates in all knock-outs. Transcriptional regulation of pathway genes will be studied under different conditions (nutrient starvations, ambient pH), and the impact of global regulators on gene expression will be examined.

镰刀菌属的物种是众所周知的严重植物病原体和几种有害真菌毒素的生产者，如伏马菌素、念珠菌素、玉米赤霉烯酮、镰刀菌素C、镰刀菌酸、比卡维林、马黄素和beauvericin。有了玉米病原菌F. verticillioides的全基因组序列，以及生物技术上重要的（生产赤霉素）真菌F. fujikuroi的预期基因组序列，我们将能够确定两种有害聚酮真菌毒素生物合成的基因（簇）：到目前为止，还没有从任何镰刀菌物种中发现moniliformin生物合成基因，只有一个fusarin C生物合成基因，即杂交聚酮合成酶/非核糖体肽合成酶（PKS/NRPS）编码基因，在镰刀菌中被鉴定出来。为了减少食品、饲料和生物技术生产的赤霉素制剂中这些真菌毒素的健康风险，我们正计划确定黄螺旋菌和fujikuroi螺旋菌中fusarin C和moniliformin生物合成的未知基因和途径。为了鉴定单毛虫素特异性PKS基因，我们将在产生单毛虫素的F. fujikuroi菌株中产生候选PKS基因的敲除突变体，并鉴定一种单毛虫素-突变体。对于fusarin C，我们将删除更多已知PKS/NRPS基因附近的推定生物合成基因，并在所有敲除中鉴定中间产物。途径基因的转录调控将在不同条件下（营养饥饿，环境pH）进行研究，并将研究全球调控对基因表达的影响。