目的 基于灵芝Ganoderma lucidum的基因组和转录组数据，挖掘灵芝C₂H₂型锌指转录因子（C₂H₂-ZFPs）基因家族成员，并分析其菌丝在茉莉酸甲酯（methyl jasmonate，MeJA）诱导后的表达模式，以期发现在灵芝的生长发育、胁迫应对及次生代谢等方面起作用的C₂H₂-ZFPs。方法 通过隐马尔可夫模型搜索和保守结构域分析，对灵芝中的C₂H₂转录因子进行全基因组鉴定，利用Peptides软件包分析C₂H₂-ZFPs的理化性质，利用IQtree构建C₂H₂锌指蛋白系统发育树，利用MEME和GSDS 2.0在线工具分析保守基序和基因结构，利用qRT-PCR测定MeJA诱导后菌丝中C₂H₂基因的相对表达量。结果 灵芝中含有20个C₂H₂型基因家族成员（GlC₂H₂.01～GlC₂H₂.20），可分为5个亚群，分布在灵芝的10条染色体上。系统发育和保守基序显示灵芝中C₂H₂转录因子具有进化上的保守性，且至少含有1个zf-C₂H₂，启动子区预测分析发现了多类激素及逆境响应顺式作用元件。转录组分析结果表明C₂H₂基因在灵芝原基期表达最高，在孢子粉中几乎不表达。MeJA诱导9 h后，菌丝中GlC₂H₂.11和GlC₂H₂.20表达显著下降，其他GlC₂H₂在处理后3～9 h呈下降趋势，12 h后恢复至正常表达水平。结论 从灵芝全基因组中系统鉴定了C₂H₂转录因子家族基因，为进一步研究C₂H₂转录因子在调控灵芝发育及次生代谢方面的生物学功能提供理论参考。

Objective Based on the genome and transcriptome data of Ganoderma lucidum, this study aims to explore the members of the C₂H₂ zinc finger proteins (C₂H₂-ZFPs) gene family in G.lucidum, and analyze their expression patterns in mycelium induced by methyl jasmonate (MeJA), in order to discover C₂H₂-ZFPs that play a role in the growth and development, stress response, and secondary metabolism of G.lucidum. Methods Genome-wide identification of C₂H₂ transcription factors in G.lucidum was carried out by Hidden Markov Model search and conserved structural domain analysis, physicochemical properties of C₂H₂-ZFPs were analyzed by Peptides software package, phylogenetic tree of C₂H₂ zinc finger proteins was constructed by using IQtree, and the conserved motifs and gene structures were analyzed by using MEME and GSDS2.0 online tools, and the relative expression of C₂H₂ gene in mycelium after MeJA induction was determined by qRT-PCR. Results G.lucidum contains 20 C₂H₂-type gene family members (GlC₂H₂.01—GlC₂H₂.20), which can be categorized into five subgroups and distributed on 10 chromosomes of G.lucidum. Phylogenetic and conserved motifs revealed that C₂H₂ transcription factors in G.lucidum are evolutionarily conserved and contain at least one zf-C₂H₂, and promoter region prediction analysis identified multiple classes of hormones and adversity-responsive cis-acting elements. Transcriptome analysis results showed that C₂H₂ gene was most highly expressed in the primary base stage of G.lucidum, and was hardly expressed in sporopollen. The expression of GlC₂H₂.11 and GlC₂H₂.20 in the mycelium was significantly decreased after 9 h of MeJA induction, and the other GlC₂H₂ showed a decreasing trend from 3 to 9 h of treatment, and then recovered to the normal expression level after 12 h. Conclusion The systematic identification of C₂H₂ transcription factor gene family from the whole genome of G. lucidum provides a theoretical reference for further research on the biological functions of C₂H₂ transcription factors in regulating the development and secondary metabolism of G. lucidum.

R286.12

浙江省自然科学基金/杭州市联合基金/重点项目（LHZSZ24H280003）;浙江省农业新品种选育重大科技专项（2021C02074，2021C02073);“十四五”国家重点研发计划项目（2023YFD1201604-1）