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首页> 外文期刊>Mycologia >Multiplex PCR analysis of fumonisin biosynthetic genes in fumonisin-nonproducing Aspergillus niger and A. awamori strains
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Multiplex PCR analysis of fumonisin biosynthetic genes in fumonisin-nonproducing Aspergillus niger and A. awamori strains

机译：不产伏马毒素的黑曲霉和泡盛曲霉菌株中伏马毒素生物合成基因的多重PCR分析

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摘要

To determine the genetic basis for loss of fumonisin B-2 (FB2) biosynthesis in FB2-nonproducing Aspergillus niger and A. awamori strains, we developed multiplex PCR primer sets to amplify fragments of eight fumonisin biosynthetic pathway (fum) genes. Fragments of all eight fum genes were amplified from FB2-producing A. niger and A. awamori strains; from FB2-nonproducing strains four amplification patterns arose in which one or more fum gene fragments were absent. Southern hybridization analysis of strains yielding patterns 2 and 3 confirmed that loss of FB2 production in A. awamori is associated with gene deletions within the fumonisin biosynthetic gene cluster. In addition, we observed a fifth multiplex amplification pattern in which all eight fum gene fragments appeared. Reverse transcription-PCR analysis of strains yielding pattern 5 showed that the expression of at least one fum gene was reduced relative to expression in FB2-producing A. niger. This suggests that in these strains loss of FB2 production is a result of structural or regulatory mutations that alter gene expression or function. These results demonstrate a diversity of genotypes within FB2-nonproducing A. niger and A. awamori populations and provide tools useful for identifying certain non-toxigenic strains for industrial or ecological applications.
机译：为了确定不产生FB2的黑曲霉和泡盛曲霉菌株中伏马菌素B-2（FB2）生物合成损失的遗传基础，我们开发了多重PCR引物组以扩增8个伏马菌素生物合成途径（fum）基因的片段。从产生FB2的黑曲霉和泡盛曲霉菌株中扩增出所有八个烟气基因的片段。从不产FB2的菌株产生了四个扩增模式，其中没有一个或多个烟熏基因片段。对产生模式2和3的菌株的Southern杂交分析证实，泡盛曲霉中FB2产生的损失与伏马菌素生物合成基因簇内的基因缺失有关。此外，我们观察到第五种多重扩增模式，其中出现了所有八个fum基因片段。产生模式5的菌株的逆转录-PCR分析表明，相对于产生FB2的黑曲霉，至少一种烟熏基因的表达降低了。这表明在这些菌株中，FB2产生的损失是改变基因表达或功能的结构或调节突变的结果。这些结果证明了在不产生FB2的黑曲霉和泡盛曲霉种群中基因型的多样性，并提供了可用于鉴定某些非毒性菌株以用于工业或生态应用的工具。

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