灵芝中C_(2)H_(2)锌指蛋白家族全基因组鉴定及表达分析

Genome-wide identification and expression analysis of C_(2)H_(2)zinc finger protein family in Ganoderma lucidum

目的基于灵芝Ganoderma lucidum的基因组和转录组数据,挖掘灵芝C_(2)H_(2)型锌指转录因子(C_(2)H_(2)-ZFPs)基因家族成员,并分析其菌丝在茉莉酸甲酯(methyl jasmonate,Me JA)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C_(2)H_(2)-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C_(2)H_(2)转录因子进行全基因组鉴定,利用Peptides软件包分析C_(2)H_(2)-ZFPs的理化性质,利用IQtree构建C_(2)H_(2)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q RT-PCR测定Me JA诱导后菌丝中C_(2)H_(2)基因的相对表达量。结果灵芝中含有20个C_(2)H_(2)型基因家族成员(Gl C_(2)H_(2).01~Gl C_(2)H_(2).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C_(2)H_(2)转录因子具有进化上的保守性,且至少含有1个zf-C_(2)H_(2),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C_(2)H_(2)基因在灵芝原基期表达最高,在孢子粉中几乎不表达。Me JA诱导9 h后,菌丝中Gl C_(2)H_(2).11和Gl C_(2)H_(2).20表达显著下降,其他Gl C_(2)H_(2)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C_(2)H_(2)转录因子家族基因,为进一步研究C_(2)H_(2)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 C_(2)H_(2)zinc finger proteins(C_(2)H_(2)-ZFPs)gene family in G.lucidum,and analyze their expression patterns in mycelium induced by methyl jasmonate(MeJA),in order to discover C_(2)H_(2)-ZFPs that play a role in the growth and development,stress response,and secondary metabolism of G.lucidum.Methods Genome-wide identification of C_(2)H_(2)transcription factors in G.lucidum was carried out by Hidden Markov Model search and conserved structural domain analysis,physicochemical properties of C_(2)H_(2)-ZFPs were analyzed by Peptides software package,phylogenetic tree of C_(2)H_(2)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 C_(2)H_(2)gene in mycelium after MeJA induction was determined by qRT-PCR.Results G.lucidum contains 20 C_(2)H_(2)-type gene family members(GlC_(2)H_(2).01-GlC_(2)H_(2).20),which can be categorized into five subgroups and distributed on 10 chromosomes of G.lucidum.Phylogenetic and conserved motifs revealed that C_(2)H_(2)transcription factors in G.lucidum are evolutionarily conserved and contain at least one zf-C_(2)H_(2),and promoter region prediction analysis identified multiple classes of hormones and adversity-responsive cis-acting elements.Transcriptome analysis results showed that C_(2)H_(2)gene was most highly expressed in the primary base stage of G.lucidum,and was hardly expressed in sporopollen.The expression of GlC_(2)H_(2).11 and GlC_(2)H_(2).20 in the mycelium was significantly decreased after 9 h of MeJA induction,and the other GlC_(2)H_(2)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 C_(2)H_(2)transcription factor gene family from the whole genome of G.lucidum provides a theoretical reference for further research on the biological functions of C_(2)H_(2)transcription factors in regulating the development and secondary metabolism of G.lucidum.