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首页> 外文OA文献 >Insertional Mutagenesis of a Fungal Biocontrol Agent Led to Discovery of a Rare Cellobiose Lipid with Antifungal Activity
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Insertional Mutagenesis of a Fungal Biocontrol Agent Led to Discovery of a Rare Cellobiose Lipid with Antifungal Activity

机译：真菌生防剂的插入诱变导致发现具有抗真菌活性的罕见纤维二糖脂质

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Insertional mutagenesis was applied for the first time to a fungal biocontrol agent, Pseudozyma flocculosa, in an attempt to obtain mutants with altered antagonistic properties. Transformants were obtained via DNA-mediated transformation. Molecular analyses of the transformants revealed that multiple copies of the plasmid were integrated in tandem at one to many chromosomal loci. The transformants were screened for their biocontrol properties using standard bioassays, and the 160 tested transformants were classified into four groups: group I mutants (22 transformants) showed a stronger antagonistic effect than the wild type (WT) while those of group II (107 transformants) had a comparable antagonistic effect; group III mutants (17 transformants) had a decreased antagonistic effect relative to WT and group IV mutants (14 transformants) had lost their biocontrol properties. Culture extracts of the mutants (group IV) and WT were analyzed and compared for the presence of active metabolites which were then separated by solid-phase extraction and purified using conventional methods. Nuclear magnetic resonance experiments and analytical studies on a metabolite specifically produced by the WT revealed the presence of 2-(2′,4′-diacetoxy-5′-carboxy-pentanoyl) octadecyl cellobioside (flocculosin), a novel glycolipid with strong antifungal properties; the production of this compound would account for the biocontrol activity of P. flocculosa.
机译：首次将插入诱变应用于真菌生物防治剂Fseudozyma flocculosa，以期获得具有改变的拮抗特性的突变体。通过DNA介导的转化获得转化体。转化子的分子分析表明，质粒的多个拷贝在一个至多个染色体基因座上串联整合。使用标准生物测定法筛选转化体的生物防治特性，将测试的160个转化体分为四组：第一组突变体（22个转化体）显示出比野生型（WT）更强的拮抗作用，而第二组突变体（107个转化体）显示出更强的拮抗作用。 ）具有可比的拮抗作用；与WT相比，III组突变体（17个转化体）具有降低的拮抗作用，而IV组突变体（14个转化体）失去了其生物防治特性。分析了突变体（IV组）和WT的培养物提取物，比较了活性代谢产物的存在，然后通过固相萃取将其分离，并使用常规方法进行纯化。 WT专门产生的代谢产物的核磁共振实验和分析研究表明，存在2-（2'，4'-二乙酰氧基-5'-羧基-戊酰基）十八烷基纤维二糖苷（絮凝酶），这是一种新型的具有强抗真菌特性的糖脂;该化合物的产生将说明絮状疟原虫的生物防治活性。

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