McCormick S P, Harris L J, Alexander N J, Ouellet T, Saparno A, Allard S, Desjardins A E
Mycotoxin Research Unit, National Center for Agricultural Utilization Research, Agricultural Research Service, U.S. Department of Agriculture, Peoria, Illinois 61604, USA.
Appl Environ Microbiol. 2004 Apr;70(4):2044-51. doi: 10.1128/AEM.70.4.2044-2051.2004.
Gibberella zeae (asexual state Fusarium graminearum) is a major causal agent of wheat head blight and maize ear rot in North America and is responsible for contamination of grain with deoxynivalenol and related trichothecene mycotoxins. To identify additional trichothecene biosynthetic genes, cDNA libraries were prepared from fungal cultures under trichothecene-inducing conditions in culture and in planta. A gene designated LH1 that was highly expressed under these conditions exhibited only moderate (59%) similarity to known trichothecene biosynthetic cytochrome P450s. To determine the function of LH1, gene disruptants were produced and assessed for trichothecene production. Gene disruptants no longer produced 15-acetyldeoxynivalenol, which is oxygenated at carbon 7 (C-7) and C-8, but rather accumulated calonectrin and 3-deacetylcalonectrin, which are not oxygenated at either C-7 or C-8. These results indicate that gene LH1 encodes a cytochrome P450 responsible for oxygenation at one or both of these positions. Despite the relatively low level of DNA and amino acid sequence similarity between the two genes, LH1 from G. zeae is the probable homologue of Tri1, which encodes a cytochrome P450 required for C-8 oxygenation in F. sporotrichioides.
禾谷镰刀菌(无性态为禾谷镰孢菌)是北美洲小麦赤霉病和玉米穗腐病的主要致病菌,会导致谷物被脱氧雪腐镰刀菌烯醇及相关单端孢霉烯族毒素污染。为了鉴定更多单端孢霉烯族毒素生物合成基因,在单端孢霉烯族毒素诱导条件下,从培养物中和植物体内的真菌培养物中制备了cDNA文库。在这些条件下高表达的一个名为LH1的基因与已知的单端孢霉烯族毒素生物合成细胞色素P450的相似性仅为中等水平(59%)。为了确定LH1的功能,构建了基因敲除体并评估其单端孢霉烯族毒素的产生情况。基因敲除体不再产生在碳7(C-7)和碳8(C-8)处被氧化的15-乙酰脱氧雪腐镰刀菌烯醇,而是积累了在C-7和C-8处均未被氧化的卡氏镰刀菌素和3-脱乙酰卡氏镰刀菌素。这些结果表明,基因LH1编码一种细胞色素P450,负责在这两个位置中的一个或两个位置进行氧化。尽管这两个基因之间的DNA和氨基酸序列相似性相对较低,但禾谷镰刀菌的LH1可能是Tri1的同源物,Tri1编码在拟分枝孢镰刀菌中C-8氧化所需的一种细胞色素P450。