茜素红诱导下偏肿革裥菌SSH-cDNA文库构建与差异表达基因筛选  

Construction of SSH-cDNA Library and Screening of Differentially Expressed Genes of Lenzites gibbosa under Alizarin Decoloration

作　　者：韩树英[1] 池玉杰[1] 张健[1] 冯连荣[1,2] HAN Shuying;CHI Yujie;ZHANG Jian;FENG Lianrong(School of Forestry,Northeast Forestry University,Harbin 150040,China;Poplar Institute of Liaoning Province,Gaizhou 115213,China)

机构地区：[1]东北林业大学林学院,哈尔滨150040 [2]辽宁省杨树研究所,盖州115213

出　　处：《吉林农业大学学报》2020年第4期386-394,共9页Journal of Jilin Agricultural University

基　　金：黑龙江省自然科学基金项目(C2016006);中央高校基本科研业务费科技平台持续发展专项(2572018CP05)。

摘　　要：为了探索白腐菌偏肿革裥菌降解染料茜素红过程中的生理机制和差异表达基因,在LNAS培养基上培养菌株,以添加茜素红为处理组,加入无菌水为对照组,采用抑制性消减杂交方法构建了在染料降解过程中的差异基因cDNA文库。经过2次差减杂交后所得的阳性克隆,除去重复序列后得到75个独立基因的ESTs序列,即为差异基因的SSH-cDNA文库。文库中显著表达的差异基因主要有编码谷氨酰胺合成酶、热激蛋白70、ɑ,β水解酶、脯氨酰氨肽酶2、MFS普通基质转运蛋白、钙网结构域蛋白、20S蛋白酶体亚基α型3、3脱氧7磷酸庚酮糖合酶、醛酮还原酶、脂肪酸2羟化酶、辅酶Ⅰ黄素氧化还原酶、细胞色素P450等基因,这些差异基因及其产物参加了茜素红的氧化降解反应、糖类和蛋白质的降解作用、应激反应和信号转导途径。此研究结果可为进一步筛选偏肿革裥菌中与染料降解性状相关的基因奠定基础。In order to explore the physiological mechanism and differentially expressed genes(DEGs)in the degradation process of anthraquinone dye alizarin red,white rot fungus Lenzites gibbosa was cultured on low nitrogen asparaginesuccinate medium(LNAS)for 7 d,alizarin red was added as the treatment group,and sterile water was added as the control group.The cDNA library of DEGs was constructed by suppression subtractive hybridization(SSH).The 75 EST sequences of independent genes,i.e.,the SSH-cDNA library of the DEGs,had been gained after the repetitive sequences were removed from the obtained positive clones by two SSHs.The significantly expressed DEGs in the library are the genes encoding glutamine synthetase,heat shock protein 70,ɑ,βhydrolase,prolyl aminopeptidase2,MFS general substrate transporter,calreticulindomaincontaining protein,20S proteasome subunit alpha type 3,3deoxy7phosphoheptulonate synthase,aldo/keto reductase,fatty acid2 hydroxylase,NADH:flavin oxidoreductase,cytochrome P450,and so on.These DEGs and their products take part in the oxidative degradation reaction of the dye alizarin red,carbohydrate and protein degradation,stress reaction and signal transduction pathways.The results of this study lay an important foundation for further screening genes related to dyes,aromatic compounds,and lignin degradation from L.gibbosa.

关 键 词：偏肿革裥菌 抑制性差减杂交 差异表达基因 氧化还原酶 热激蛋白 

分 类 号：S718.81􀆰[农业科学—林学] Q78[生物学—分子生物学]