木霉菌T23胶毒素合成基因的生物信息学分析与克隆

胶毒素是生防木霉菌重要的次生代谢产物之一。本研究以生防木霉菌T23为供试材料,旨在通过生物信息学技术及表达分析,挖掘木霉菌T23中胶毒素合成候选基因,探索木霉菌胶毒素合成的分子调控机制,可为新型生物农药的开发及应用提供理论依据。研究表明,木霉菌T23中胶毒素合成候选基因簇全长28 kb,簇内包含了8个基因,分别与烟曲霉胶毒素合成基因簇内的gliP、gliC、gliN、gliK、gliI、gliG、gliF、gliM高度同源。提取培养2 d、3 d、4 d、5 d的木霉菌T23菌丝的RNA,通过半定量RT-PCR技术探索各候选基因在木霉菌T23不同生长时期的表达情况,显示各基因在不同生长时期均有表达,属于组成型表达基因。成功克隆得到木霉菌T23中的gliP-T23基因并完成基因结构分析,该基因全长6 339 bp,由4个外显子和3个内含子组成,为后续的基因功能验证提供基础。

Bioinformatic Analysis and Cloning of Gliotoxin Biosynthetic Gene Cluster of Trichoderma T23

Gliotoxin is one of the important secondary metabolites of bio-control Trichoderma spp.In this paper,using bio-control Trichoderma T23 as tested material,gliotoxin biosynthetic candidate gene cluster in Trichoderma T23 was identified and the molecular regulation mechansim of Trichoderma gliotoxin biosynthesis was explored through bioinformatics technique and expression analysis,which could provide theoretical basis for the development and application of new biopesticides.The results showed that the full length of gliotoxin biosynthesis candidate gene cluster in Trichoderma T23 was 28 kb,including 8 genes,which were highly homologous to gliP,gliC,gliN,gliK,gliI,gliG,gliF,gliM located in gliotoxin biosynthesis gene cluster of Aspergillus fumigatus,respectively.The RNAs of Trichoderma T23 mycelium after 2 d,3 d,4 d and 5 d cultivation were extracted and the expression of each candidate gene in the different growth stages of Trichoderma T23 was explored through semi-quantitative RT-PCR.The result showed that each gene expressed in different growth stages,belonging to constitutive expression genes.gliP-T23 in Trichoderma T23 was cloned and obtained,and gene structure analysis was completed.The full length of gliP-T23 was 6 339 bp,including 4 exons and 3 introns,which could provide the basis for subsequent genetic function verification.