Jiangsu Key Laboratory for Microbes and Microbial Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, College of Life Science, Nanjing Normal Universitygrid.260474.3, Nanjing, People's Republic of China.
College of Life Science and Chemistry, Jiangsu Second Normal University, Nanjing, People's Republic of China.
mBio. 2021 Feb 22;13(1):e0362621. doi: 10.1128/mbio.03626-21. Epub 2022 Feb 1.
Coprinopsis cinerea has seven homologs of the Aspergillus nidulans transcription factor NsdD. Of these, CcNsdD1 and CcNsdD2 from show the best identities of 62 and 50% to A. nidulans NsdD, respectively. After 4 days of constant darkness cultivation, , but not , was upregulated on the first day of light/dark cultivation to induce fruiting bodies, and overexpression of , but not , produced more fruiting bodies under a light/dark rhythm. Although single knockdown of did not affect fruiting body production due to upregulation of its homolog , the double-knockdown CcNsdD1/NsdD2-RNAi transformant showed defects in fruiting body formation under a light/dark rhythm. Knockdown of / led to the differentiation of primary hyphal knots into sclerotia rather than secondary hyphal knots under a light/dark rhythm, similar to the differentiation of primary hyphal knots into sclerotia of the wild-type strain under darkness. The CcNsdD2-overexpressing transformant produced more primary hyphal knots, secondary hyphal knots, and fruiting bodies under a light/dark rhythm but only more primary hyphal knots and sclerotia under darkness. RNA-seq revealed that some genes reported previously to be involved in formation of hyphal knots and primordia, cyclopropane-fatty-acyl-phospholipid synthases , galectins , and hydrophobins were downregulated in the CcNsdD1/NsdD2-RNAi transformant compared to the mock transformant. ChIP-seq and electrophoretic mobility shift assay demonstrated that CcNsdD2 bound to promoter regulatory sequences containing a GATC motif in , , , and . A molecular mechanism by which CcNsdD2 regulates the developmental fate of under dark or light conditions is proposed. The model mushroom Coprinopsis cinerea exhibits remarkable photomorphogenesis during fruiting body development. This study reports that the transcription factor CcNsdD2 promotes primary hyphal knot formation by upregulating , , , and . Although the induction of is not under direct control of light and photoreceptors, the CcNsdD2-mediated developmental fates of the primary hyphal knots depend on the following light/dark rhythm cultivation or dark cultivation after full growth of mycelia in the constant dark cultivation. This study provides new insight into the molecular mechanism by which CcNsdD2 regulates the developmental fate of under dark or light conditions. In addition, the result that overexpression of induced more secondary hyphal knots, primordia, and fruiting bodies under light/dark rhythm cultivation conditions has potential applied value in the edible mushroom industry.
灰盖鬼伞有七个与构巢曲霉转录因子 NsdD 同源的蛋白。其中，来自 的 CcNsdD1 和 CcNsdD2 与构巢曲霉 NsdD 的同一性最好，分别为 62%和 50%。在连续黑暗培养 4 天后， 但不是 ，在光照/黑暗培养的第一天被上调以诱导产生子实体，并且在光照/黑暗节律下过表达 会产生更多的子实体。尽管由于同源物的上调，单敲低 不会影响子实体的产生 ，但 CcNsdD1/NsdD2-RNAi 敲低转化体在光照/黑暗节律下表现出子实体形成缺陷。敲低 / 导致初级菌丝结分化为菌核而不是次级菌丝结，类似于黑暗条件下野生型菌株初级菌丝结分化为菌核。CcNsdD2 过表达转化体在光照/黑暗节律下产生更多的初级菌丝结、次级菌丝结和子实体，但在黑暗条件下仅产生更多的初级菌丝结和菌核。RNA-seq 显示，一些先前报道的与菌丝结和原基形成有关的基因，如环丙烷脂肪酸磷脂合成酶、半乳凝集素和疏水性蛋白 在 CcNsdD1/NsdD2-RNAi 转化体中与对照转化体相比下调。ChIP-seq 和电泳迁移率变动分析显示，CcNsdD2 结合到含有 GATC 基序的启动子调节序列中， 、 、 和 。提出了 CcNsdD2 在黑暗或光照条件下调节 发育命运的分子机制。模式蘑菇灰盖鬼伞在子实体发育过程中表现出显著的光形态发生。本研究报告，转录因子 CcNsdD2 通过上调 、 、 和 来促进初级菌丝结的形成。尽管 的诱导不受光和光受体的直接控制，但 CcNsdD2 介导的初级菌丝结发育命运取决于以下光照/黑暗节律培养或在恒定黑暗培养中菌丝完全生长后的黑暗培养。本研究为 CcNsdD2 在黑暗或光照条件下调节 发育命运的分子机制提供了新的见解。此外，过表达 在光照/黑暗节律培养条件下诱导产生更多的次级菌丝结、原基和子实体的结果在食用菌工业中具有潜在的应用价值。
