白粉菌诱导月季对甜菜夜蛾产生抗性的关键候选基因筛选及CYP71的生物信息学分析

Candidate Genes Selection in Powdery Mildew-induced Resistance of Rose Plant to Beet Armyworms and the Bioinformatics Analysis on CYP71

【目的】通过检测白粉菌侵染月季后转录组基因的表达量变化,筛选白粉菌诱导月季产生对甜菜夜蛾抗性的关键候选基因,为月季抗虫基因的功能研究及抗虫品种的培育提供参考。【方法】以月季品种艳粉(Rosa chinensis Jacq.)为研究材料,利用二代测序技术对白粉菌侵染月季前后基因的表达量进行转录组测序。利用RSEM对基因的表达水平进行定量分析,DESeq2软件进行表达差异分析,利用KOBAS将得到的差异表达基因进行KEGG富集分析。对筛选出的可能参与三萜类物质生物合成的CYP71D差异表达基因(Differentially expressed genes,DEGs)进行生物信息学分析,并分析了CYP71D蛋白与抗虫性的关系。【结果】转录组测序共检测到差异表达基因1 646个,共643个差异表达基因被富集到81条KEGG通路中,其中31个差异表达基因与萜类合成相关。生物信息学分析结果表明了CYP71D亚家族的物种特异性。在植物中CYP71A主要参与类单萜的生物合成,CYP71D主要参与单萜类、倍半萜类、二萜类、三萜类等与抗病、虫有关的次生代谢物的生物合成。【结论】月季细胞色素P450超家族蛋白酶中CYP71簇的CYP71A、 CYP71D亚家族可能与三萜化合物Dehydro(11,12)ursolicacid lactone(DUA)的生物合成有关,表明CYP71D家族蛋白参与修饰三萜化合物的底物识别位点,本研究为探索寄主植物介导的间接的病虫互作机理的研究提供了一定的思路。

【Objective】 The transcriptomes of rose plants artificially infected by powdery mildew were obtained to identify the genes responsible for the resistance to infestation by beet armyworms. 【Method】 Transcriptomes of Yanfen rose(Rosa chinensis Jacq.) before and after being infected by powdery mildew were obtained using the second-generation sequencing technology. RSEM was used to quantitatively analyze the gene expressions, DESeq2 software for the expression differences,and KOBAS for the KEGG enrichment analysis on the differentially expressed genes(DEGs). Bioinformatics of the DEGs of CYP71D susceptible of associating with triterpenoids biosynthesis was secured to study the relationship between the CYP71D and the insect resistance. 【Result】 A total of 1 646 DEGs were detected by the transcriptome sequencing. Of which, 643were enriched into 81 KEGG pathways with 31 DEGs related to terpenoid synthesis. The bioinformatics on them showed a species specificity of CYP71D subfamily. In plants, CYP71A were mainly involved in the biosynthesis of monoterpenes, while CYP71D in that of the disease and insect resistance-related secondary metabolites, such as monoterpenes, sesquiterpenes,diterpenes, and triterpenoids. 【Conclusion】 The CYP71A and CYP71D subfamilies of the cytochrome P450 enzyme in the rose might relate to the synthesis of dehydro(11,12) ursolic acid lactone(DUA). It suggested a possible association of CYP71D with the formation of triterpenoids. The results provided information for the study on the relationship between the genes in host plants and infecting insects.