疏水蛋白基因Cmhyd4对蛹虫草生长发育及基因表达的调控OA
Regulation of hydrophobin gene Cmhyd4 on the growth,development and gene expression of Cordyceps militaris
疏水蛋白为丝状真菌特有的小分子分泌蛋白,疏水蛋白基因调控生长发育的机制缺乏深入研究.前期通过基因编辑获得了蛹虫草疏水蛋白基因Cmhyd4缺失株(∆Cmhyd4cas9),本研究对该基因缺失前后蛹虫草菌丝生长和子实体发育进行比较分析,发现 Cmhyd4基因缺失促进蛹虫草气生菌丝生长,提高单瓶子实体数目和生物学转化率.以蛹虫草野生型(WT)菌株和∆Cmhyd4cas9 菌株为实验材料,取其菌丝(MY)、原基(PR)、成熟子实体(MF)阶段样品进行比较转录组学分析.结果显示,相比WT菌株,Cmhyd4的缺失影响各阶段基因转录水平,PR阶段差异基因数目最多(349个),其次为 MY阶段(179个),MF阶段数目最少(93个);疏水蛋白家族成员基因不同发育阶段转录水平为其功能提供了关键解释,Cmhyd4 的缺失改变了家族其他基因转录水平,表现出显著的基因补偿效应.GO分析结果显示,Cmhyd4缺失后的差异基因在MY阶段主要富集在细胞周期过程、有丝分裂周期等过程,PR 和 MF 阶段主要富集在非生物胁迫响应等相关过程;与 GO 富集结果一致,Cmhyd4 缺失后子实体抗 NaCl 胁迫能力显著提升,而菌丝阶段无明显变化.RT-qPCR 结果证实,Cmhyd4的缺失并未改变渗透压胁迫响应信号通路中 4 个基因在 MY 阶段的表达,而显著提高其在PR和MF阶段的表达.本研究全面展示了Cmhyd4对蛹虫草生长发育的影响及对基因表达的调控,为后续蛹虫草分子育种提供基因资源.
Hydrophobins,small secreted proteins unique to filamentous fungi,play critical roles in fungal growth and development yet their functional mechanisms are poorly understood.Previously,a hydrophobin gene Cmhyd4 knockout strain ∆Cmhyd4cas9 of Cordyceps militaris was generated via gene editing.A comparative analysis of mycelial growth and fruiting body development before and after Cmhyd4 deletion was conducted.It was revealed that the loss of Cmhyd4 promoted aerial hyphae growth,and increased the number of fruiting bodies and biological conversion efficiency per cultivation bottle.Using wild-type(WT)and ∆Cmhyd4cas9 strains as experiment materials,samples from the mycelia(MY),primordium(PR),and mature fruiting body(MF)were subjected to comparative transcriptomic analysis.The results showed that Cmhyd4 deletion altered gene transcript levels across all stages as compared with WT strain.The highest differentially expressed genes(DEGs)occurred at PR stage(349),and MY stage(179)was the next,while the fewest at MF stage(93).The transcriptional patterns of hydrophobin family genes at different developmental stages provided critical insights into their functions.The deletion of Cmhyd4 altered the transcription levels of the other family members,demonstrating significant gene compensation effects.GO enrichment analysis revealed that after Cmhyd4 gene deletion,DEGs at the MY stage were primarily associated with cell cycle and mitosis,whereas DEGs at PR and MF stages were linked to abiotic stress response.Consisted with GO analysis,the ∆Cmhyd4cas9 strain exhibited significant enhancement of NaCl stress resistance in fruiting bodies,while no notable changes were observed in the mycelial stage.RT-qPCR validation confirmed that four genes in the osmotic stress response pathway showed no differential expression at MY stage after Cmhyd4 gene deletion,but significant upregulation occurred in PR and MF stages.This study comprehensively elucidates the impacts of Cmhyd4 on the growth and development of C.militaris,along with its regulatory role in gene expression,providing genetic resources for future molecular breeding of C.militaris.
任慧慧;王佳媛;李忞阳;刘苏雅;张婕;马敖;李守勉;李肖
河北农业大学园艺学院 河北省蔬菜种质创新与利用重点实验室 河北省蔬菜产业协同创新中心,河北 保定 071001河北农业大学园艺学院 河北省蔬菜种质创新与利用重点实验室 河北省蔬菜产业协同创新中心,河北 保定 071001河北农业大学园艺学院 河北省蔬菜种质创新与利用重点实验室 河北省蔬菜产业协同创新中心,河北 保定 071001河北农业大学园艺学院 河北省蔬菜种质创新与利用重点实验室 河北省蔬菜产业协同创新中心,河北 保定 071001河北农业大学园艺学院 河北省蔬菜种质创新与利用重点实验室 河北省蔬菜产业协同创新中心,河北 保定 071001河北农业大学园艺学院 河北省蔬菜种质创新与利用重点实验室 河北省蔬菜产业协同创新中心,河北 保定 071001河北农业大学园艺学院 河北省蔬菜种质创新与利用重点实验室 河北省蔬菜产业协同创新中心,河北 保定 071001河北农业大学园艺学院 河北省蔬菜种质创新与利用重点实验室 河北省蔬菜产业协同创新中心,河北 保定 071001
蛹虫草菌丝子实体非生物胁迫基因
Cordyceps militarismyceliumfruiting bodyabiotic stressgene
《菌物学报》 2026 (6)
276-289,14
国家自然科学基金(32202569)河北省自然科学基金(C2023204076)河北农业大学创新创业训练计划项目(2026070)河北省科技计划项目重点研发计划(21326315D)河北省省属高等学校基本科研业务费研究项目(KY2023055)河北农业大学引进人才科研专项(YJ202249) This work was supported by the National Natural Science Foundation of China(32202569),the Natural Science Foundation of Hebei Province(C2023204076),the Innovation and Entrepreneurship Training Program Project of Hebei Agricultural University(2026070),the Key Research and Development Planning Project in Science and Technology of Hebei Province(21326315D),the Research Project of Basic Scientific Research Expenses of Colleges and Universities in Hebei Province(KY2023055),and the Talent Introduction Scientific Research Special Project of Hebei Agricultural University(YJ202249).
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