A WD40 Protein Encoding Gene Fvcpc2 Positively Regulates Mushroom Development and Yield in Flammulina velutipes
A WD40 Protein Encoding Gene Fvcpc2 Positively Regulates Mushroom Development and Yield in Flammulina velutipes
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DOI：
10.3389/fmicb.2020.00498
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发表时间：
2020-03
影响因子：
5.2
通讯作者：
Taju Wu;Zhen-ying Zhang;Chengcheng Hu;Long Zhang;Shenglong Wei;Shaojie Li
中科院分区：
文献类型：
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作者：
Taju Wu;Zhen-ying Zhang;Chengcheng Hu;Long Zhang;Shenglong Wei;Shaojie Li
子囊菌门和担子菌门是两个密切相关的门，两个门的真菌在子实体形成过程中具有一些共同的形态发育过程。在粗糙脉孢菌中，先前报道了具有七个 WD40 重复结构域的 Gβ 样蛋白 CPC-2。通过将来自子囊菌门和担子菌门的7种不同真菌物种的CPC-2直系同源编码基因转化到粗糙脉孢菌的cpc-2缺失突变体中，我们证明所有测试的CPC-2直系同源基因都能够弥补cpc-2缺失突变体在性发育中的缺陷，表明来自子囊菌门和担子菌门的CPC-2蛋白具有相似的细胞功能。使用金针菇作为蘑菇物种的模型系统，对 CPC-2 直系同源物 FvCPC2 进行了表征。 Fvcpc2 在子实体发育过程中增加转录。 RNAi 敲低 Fvcpc2 完全损害了子实体的形成。在三个 Fvcpc2 敲低突变体中，编码腺苷酸环化酶和蛋白激酶 A 催化亚基的基因的转录水平显着降低，并且集落生长变得比野生型慢。在培养基中添加 cAMP 或 PKA 激活剂 8-Bromo-cAMP 可将 Fvcpc2 敲低突变体恢复至野生型集落生长表型，表明 cAMP 的产生参与 FvCPC2 的调节机制。 Fvcpc2的敲除还削弱了一些与子实体发育相关的基因对性发育诱导的转录反应，包括4个jacalin相关的凝集素编码基因、4个疏水蛋白编码基因和3个功能未知基因，表明这些基因参与了FvCPC2调节子实体发育的机制。与野生型相比，所有三个 Fvcpc2 过表达菌株均表现出蘑菇产量增加和培养时间缩短，这表明 Fvcpc2 可以成为冬蘑菇育种的有前途的参考基因。由于FvCPC2的直系同源基因在不同食用菌的子实体发育过程中高度保守且特异表达，因此其他蘑菇物种中编码FvCPC2直系同源基因的基因在育种中也具有潜在的应用。
Ascomycota and Basidiomycota are two closely related phyla and fungi in two phyla share some common morphological developmental process during fruiting body formation. In Neurospora crassa, the Gβ-like protein CPC-2 with a seven-WD40 repeat domain was previously reported. By transforming CPC-2 ortholog encoding genes, from 7 different fungal species across Ascomycota and Basidiomycota, into the cpc-2 deletion mutant of N. crassa, we demonstrate that all tested CPC-2 ortholog genes were able to complement the defects of the cpc-2 deletion mutant in sexual development, indicating that CPC-2 proteins from Ascomycota and Basidiomycota have the similar cellular function. Using Flammulina velutipes as a model system for mushroom species, the CPC-2 ortholog FvCPC2 was characterized. Fvcpc2 increased transcription during fruiting body development. Knockdown of Fvcpc2 by RNAi completely impaired fruiting body formation. In three Fvcpc2 knockdown mutants, transcriptional levels of genes encoding adenylate cyclase and protein kinase A catalytic subunit were significantly lower and colony growth became slower than wild type. The addition of cAMP or the PKA-activator 8-Bromo-cAMP into the medium restored the Fvcpc2 knockdown mutants to the wild-type colony growth phenotype, suggesting that the involvement of cAMP production in the regulatory mechanisms of FvCPC2. Knockdown of Fvcpc2 also weakened transcriptional responses to sexual development induction by some genes related to fruiting body development, including 4 jacalin-related lectin encoding genes, 4 hydrophobin encoding genes, and 3 functionally-unknown genes, suggesting the participation of these genes in the mechanisms by which FvCPC2 regulates fruiting body development. All three Fvcpc2 overexpression strains displayed increased mushroom yield and shortened cultivation time compared to wild type, suggesting that Fvcpc2 can be a promising reference gene for Winter Mushroom breeding. Since the orthologs of FvCPC2 were highly conserved and specifically expressed during fruiting body development in different edible mushrooms, genes encoding FvCPC2 orthologs in other mushroom species also have potential application in breeding.