桑黄Zn(Ⅱ)2Cys6锌簇蛋白转录因子及其在不同碳氮源条件下的表达  

Zn(Ⅱ)2Cys6-type transcription factors in Sanghuangporus sanghuang grown under different carbon and nitrogen sources

作　　者：张雪蕊 张子蕴 王毅[2] 原晓龙[2] 杨焱[3] ZHANG Xue-Rui;ZHANG Zi-Yun;WANG Yi;YUAN Xiao-Long;YANG Yan(School of Life Sciences,Ludong University,Yantai,Shandong 264025,China;Laboratory of Forest Plant Cultivation and Utilization,Yunnan Academy of Forestry and Grassland and The Key Laboratory of Rare and Endangered Forest Plants of State Forestry Administration,Kunming,Yunnan 650201,China;Istitute of Edible Fungi,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China)

机构地区：[1]鲁东大学生命科学学院,山东烟台264025 [2]云南省林业与草原科学院云南省森林植物培育与开发利用重点实验室,国家林业局云南珍稀濒特森林植物保护和繁育重点实验室,云南昆明650201 [3]上海市农业科学院食用菌研究所,上海201106

出　　处：《菌物学报》2021年第7期1676-1687,共12页Mycosystema

基　　金：国家自然科学基金(31860177);分子生物学研究团队(LKYTD-2020-001);云南省重点研发计划-农业领域“云南松林分质量精准提升关键技术研发应用”(2018BB006)。

摘　　要：Zn(Ⅱ)2Cys6锌簇蛋白转录因子(C6zinc)在真菌次生代谢产物合成中发挥重要作用。本研究首先利用本地BLAST,从桑黄Sanghuangporussanghuang全基因组中获得Zn(Ⅱ)2Cys6锌簇蛋白转录因子编码基因,并利用生物信息学手段分析编码蛋白保守结构域、一级结构及二级结构,构建蛋白系统发育树,最后利用半定量PCR对它们在不同碳源和氮源培养条件下的表达情况进行检测。结果显示:从桑黄基因组中分析获得的11个Zn(Ⅱ)2Cys6锌簇蛋白均具有Cy6型锌指基序,属于GAL4型锌簇蛋白转录因子;它们均为不稳定亲水性蛋白,具磷酸化修饰位点,糖基化修饰位点较少或没有,定位于细胞核中;其二级结构主要以无规卷曲和α螺旋为主;系统发育树分析结果显示,桑黄Zn(Ⅱ)2Cys6锌簇蛋白分为2个大分支,其中Ⅰ类分支转录因子的保守结构域分布于蛋白N-端,Ⅱ类分支转录因子的保守结构域则分布于蛋白C-端;11个桑黄Zn(Ⅱ)2Cys6锌簇蛋白转录因子在不同培养基培养的菌丝体中呈现差异化表达,其中,肌醇培养基和乳糖培养基能够有效促进大部分锌簇蛋白转录因子的表达;另外,基因簇分析显示桑黄锌簇蛋白SHCZ4可能是NRPS-PKS杂合基因簇体系的途经特异性转录因子。该结果将为桑黄次生代谢产物合成调控相关转录因子的研究以及潜在次生代谢相关基因簇的挖掘提供参考依据。Zn(Ⅱ)2Cys6 transcription factor(C6 zinc)plays a significant role in the synthesis of secondary metabolites in fungi.In this study eleven genes that encode C6 zinc from the whole Sanghuangporus sanghuang genome were identified by using local BLAST.Bioinformatics analysis method were utilized to analyse the conserved domains as well as the primary and secondary structures of protein products.Protein phylogenetic tree was constructed to classify the C6 zinc in S.sanghuang.Besides,their gene expressions under different carbon source and nitrogen source culture conditions were analyzed by semi-quantitative PCR.The result shows that the eleven S.sanghuang C6 zinc proteins are unstable and hydrophilic,with phosphorylated modification sites,but without glycosylation modification sites or signal peptides,and all of them locate in the nucleus.The secondary structure is mainly composed of random coil and alpha helix.These S.sanghuang C6 zinc proteins were classified into two main branches in phylogenetic trees.Among them,the conserved domains of the group Ⅰ proteins are distributed at the N-terminal,and the others in branch Ⅱ are distributed at the C-terminal.Eleven S.sanghuang C6 zinc transcription factors were differentially expressed in mycelia cultured on different media,and inositol medium as well as lactose medium could effectively promote the expression of most of them.The preliminary analysis of the gene cluster suggests that SHCZ4 might be a pathway-specific transcription factor of NRPS-PKS hybrid metabolites.These results will provide a reference for the study of transcription factors related to the synthesis of secondary metabolites and for the mining of potential secondary metabolic-related gene clusters in S.sanghuang.

关 键 词：桑黄 Zn(Ⅱ)2Cys6锌簇蛋白转录因子 次生代谢产物 基因簇 

分 类 号：S567.3[农业科学—中草药栽培]