Characterization of the Podospora anserina (Rabenh.) Niessl Peroxidase Gene Family

R. Ferrari, Valérie Gautier, N. Tangthirasunun, Laetitia Chan Ho Tong, Philippe Silar
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Abstract

Major role of peroxidases in plant biomass degradation is well-established in the white rot basidiomycetes. On the contrary, peroxidases are not used for this purpose by brown rot basidiomycetes, which use instead a non-enzymatic mechanism. In the case of the ascomycetes, not much is known although these fungi have peroxidase genes. Here, we identify and characterize the peroxidase genes of Podospora anserina (Rabenh.) Niessl, an ascomycete used to study development and lignocellulose degradation. We show that this fungus has one class II peroxidase, one hybrid B peroxidase, one haloperoxidase, four functional aromatic peroxygenases, one glutathione peroxidase, one cytochrome C peroxidase and one alkyl peroxidase, but lacks a dye peroxidase. We show that potentially secreted peroxidases (i.e., the class II, hybrid B, haloperoxidase and aromatic peroxygenase peroxidases) present a patchy phylogenetic distribution compatible with an accessory role in finely adapting the different fungal species to their ecological niche, rather than being involved in fundamental roles in fungal biology. Accordingly, targeted gene deletions of the different P. anserina peroxidase genes identified only one phenotype, seemingly an alteration of the timing of ascospore maturation at intermediate concentration of vanillic acid. However, direct measure of peroxidase activity did not show drastic loss of activity in the tested mutants, suggesting compensation between the enzymes. Hence, in P. anserina , peroxidases appear to have a minor role in biomass degradation, unlike what has been described in white rot fungi, and in this regard P. anserina appears to be similar to brown rot fungi.
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Podospora anserina (Rabenh.) Niessl 过氧化物酶基因家族的特征描述
过氧化物酶在植物生物量降解过程中的主要作用已在白腐基枝菌中得到证实。相反,褐腐基枝菌不使用过氧化物酶,而是使用一种非酶机制。至于子囊菌,虽然这些真菌都有过氧化物酶基因,但人们对它们的了解并不多。在这里,我们鉴定并描述了 Podospora anserina (Rabenh.) Niessl 的过氧化物酶基因,这是一种用于研究发育和木质纤维素降解的子囊菌。我们发现该真菌有一个 II 类过氧化物酶、一个杂交 B 过氧化物酶、一个卤过氧化物酶、四个功能性芳香族过氧化物酶、一个谷胱甘肽过氧化物酶、一个细胞色素 C 过氧化物酶和一个烷基过氧化物酶,但缺乏染料过氧化物酶。我们的研究表明,潜在的分泌型过氧化物酶(即 II 类、杂交 B、卤过氧化物酶和芳香族过氧化物酶过氧化物酶)在系统发育中呈斑块状分布,这与它们在使不同真菌物种精细地适应其生态位方面所起的辅助作用相吻合,而不是参与真菌生物学的基本作用。因此,对不同 P. anserina 过氧化物酶基因的定向基因缺失只发现了一种表型,似乎是在香草酸中间浓度下改变了腹孢子成熟的时间。然而,对过氧化物酶活性的直接测量并未显示出测试突变体中活性的急剧下降,这表明酶之间存在补偿作用。因此,在鹅膏菌中,过氧化物酶似乎在生物质降解过程中作用不大,这与白腐真菌中的情况不同,在这方面鹅膏菌似乎与褐腐真菌相似。
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