灰葡萄孢Bctre1的基因组定位、蛋白质结构预测及表达分析

Genome Localization, Protein Structure Prediction and Expression Analysis of Bctre1 in Botrytis cinerea

本研究选取灰葡萄孢菌株BC05.10为试材,克隆Bctre1基因的核苷酸序列并对其进行生物信息学分析,同时采用荧光定量PCR法研究该基因在不同侵染时期、分生孢子萌发过程中以及不同碳源、海藻糖诱导下的表达情况。结果表明:该基因ID号为5428296,全长3341bp,位于scaffold_17负链的495880~499211位置,与核盘菌Sstre1的同源关系较近;BcTRE1蛋白由1033个氨基酸残基编码组成,该蛋白N端具有2个6发卡结构的超糖苷酶家族和1个C端糖基水解酶家族63。荧光定量PCR结果显示,随着侵染时间的延长Bctre1表达量显著增加(P<0.05),48h后表达量增加不显著;Bctre1在分生孢子萌发24h时表达量最高;Bctre1在海藻糖为单一碳源的培养基中表达量最高,在海藻糖诱导下12h后Bctre1表达量显著增加(P<0.05)。综上,Bctre1与侵染寄主过程中分生孢子萌发及不同碳源利用密切相关,在侵染后期对促进菌丝生长发挥重要作用。

In this study,the Botrytis cinerea strain BC 05.10 was selected as test material,and the nucleotide sequence of Bctre1 gene were cloned and analyzed by bioinformatics method.The expression of Bctre1 gene at different infection stages,in the process of conidia germination,and under different carbon sources and trehalose induction were studied by fluorescence quantitative PCR method.The results showed that the gene ID number was 5428296,and its total length was 3341 bp,which was located at 495880~499211 sites of the negative chain of scaffold17.The homologous relationship between Bctre1 and Sstre1 was close.The protein encoded by Bctre1 gene contained 1033 amino acid residues and had two 6-hairpin structure super glucosidase family and one C-terminal glycosyl hydrolase family 63.The results of fluorescence quantitative PCR showed that the expression of Bctre1 increased significantly with the increase of infection time(P<0.05),while the increase of Bctre1 expression after 48 h was not significant.The expression of Bctre1 was the highest when conidia germinated for 24 h and the trehalose was used as the single carbon source in medium,and it increased significantly after 12 h under trehalose induction(P<0.05).In summary,Bctre1 was closely related to conidia germination and different carbon source utilization during the process of infecting host,and played an important role in promoting mycelial growth at late stage of infection.