The divergence of DHN-derived melanin pathways in Metarhizium robertsii

Linan Xie, Yang Liu, Yujie Zhang, Kang Chen, Qun Yue, Chen Wang, Baoqing Dun, Yuquan Xu, Liwen Zhang
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Abstract

The important role of dihydroxynaphthalene-(DHN) melanin in enhancing fungal stress resistance and its importance in fungal development and pathogenicity are well-established. This melanin also aids biocontrol fungi in surviving in the environment and effectively infecting insects. However, the biosynthetic origin of melanin in the biocontrol agents, Metarhizium spp., has remained elusive due to the complexity resulting from the divergence of two DHN-like biosynthetic pathways. Through the heterologous expression of biosynthetic enzymes from these two pathways in baker’s yeast Saccharomyces cerevisiae, we have confirmed the presence of DHN biosynthesis in M. roberstii, and discovered a novel naphthopyrone intermediate, 8, that can produce a different type of pigment. These two pigment biosynthetic pathways differ in terms of polyketide intermediate structures and subsequent modification steps. Stress resistance studies using recombinant yeast cells have demonstrated that both DHN and its intermediates confer resistance against UV light prior to polymerization; a similar result was observed for its naphthopyrone counterpart. This study contributes to the understanding of the intricate and diverse biosynthetic mechanisms of fungal melanin and has the potential to enhance the application efficiency of biocontrol fungi such as Metarhizium spp. in agriculture.

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Metarhizium robertsii 中 DHN 衍生黑色素途径的分化
二羟基萘(DHN)黑色素在增强真菌抗逆性方面的重要作用及其在真菌发育和致病性方面的重要性已得到公认。这种黑色素还有助于生物控制真菌在环境中生存并有效感染昆虫。然而,由于两种类似于 DHN 的生物合成途径的分化所导致的复杂性,黑色素在生防真菌 Metarhizium spp.通过在面包酵母中异源表达这两条途径中的生物合成酶,我们证实了罗伯茨菌中存在 DHN 生物合成,并发现了一种新型萘醌中间体 8,它可以产生一种不同类型的色素。这两种色素的生物合成途径在多酮中间体结构和后续修饰步骤方面有所不同。利用重组酵母细胞进行的抗应激研究表明,DHN 及其中间体在聚合之前都具有抗紫外线的能力;其对应的萘并吡喃酮也有类似的结果。这项研究有助于人们了解真菌黑色素复杂多样的生物合成机制,并有可能提高 Metarhizium 等生物控制真菌在农业中的应用效率。
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