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Functional Analysis of Developmentally Regulated Genes chs7 and sec22 in the Ascomycete Sordaria macrospora

机译:子囊夜蛾的发育调控基因chs7和sec22的功能分析

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摘要

During sexual development, filamentous ascomycetes form complex, three-dimensional fruiting bodies for the generation and dispersal of spores. In previous studies, we identified genes with evolutionary conserved expression patterns during fruiting body formation in several fungal species. Here, we present the functional analysis of two developmentally up-regulated genes, chs7 and sec22, in the ascomycete Sordaria macrospora. The genes encode a class VII (division III) chitin synthase and a soluble N-ethylmaleimide-sensitive-factor attachment protein receptor (SNARE) protein, respectively. Deletion mutants of chs7 had normal vegetative growth and were fully fertile but showed sensitivity toward cell wall stress. Deletion of sec22 resulted in a reduced number of ascospores and in defects in ascospore pigmentation and germination, whereas vegetative growth was normal in the mutant. A SEC22-EGFP fusion construct under control of the native sec22 promoter and terminator regions was expressed during different stages of sexual development. Expression of several development-related genes was deregulated in the sec22 mutant, including three genes involved in melanin biosynthesis. Our data indicate that chs7 is dispensable for fruiting body formation in S. macrospora, whereas sec22 is required for ascospore maturation and germination and thus involved in late stages of sexual development.
机译:在性发育过程中,丝状子囊菌形成了复杂的三维子实体,用于孢子的产生和散布。在以前的研究中,我们鉴定了几种真菌物种在子实体形成过程中具有进化保守表达模式的基因。在这里,我们介绍了在子囊拟南芥中的两个发育上调的基因chs7和sec22的功能分析。这些基因分别编码VII类(III类)几丁质合酶和可溶性N-乙基马来酰亚胺敏感性因子附着蛋白受体(SNARE)蛋白。 chs7的缺失突变体具有正常的营养生长,并且完全可育,但对细胞壁胁迫表现出敏感性。 sec22的删除导致子囊孢子数量的减少以及子囊孢子色素沉着和发芽的缺陷,而该突变体的营养生长正常。在性发育的不同阶段表达了受天然sec22启动子和终止子区域控制的SEC22-EGFP融合构建体。在sec22突变体中,一些发育相关基因的表达被下调,包括涉及黑色素生物合成的三个基因。我们的数据表明chs7对于大孢链霉菌子实体的形成是必不可少的,而sec22是子囊孢子成熟和萌发所必需的,因此参与性发育的后期。

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