山楂属植物中苹果褪绿叶斑病毒全基因组解析及其致病机制研究

项目名称： 山楂属植物中苹果褪绿叶斑病毒全基因组解析及其致病机制研究

项目编号： No.31470678

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 代红艳

作者单位： 沈阳农业大学

项目金额： 88万元

中文摘要： 苹果褪绿叶斑病毒（Apple chlorotic leaf spot virus, ACLSV）是一种广泛侵染蔷薇科植物的RNA病毒，对经济性状具有严重影响，但其致病机制并不明确。山楂原产我国，是蔷薇科中重要的经济林植物和良好的园林观赏植物。前期研究结果表明，我国的山楂属植物中宿有ACLSV。本项目拟以国家果树种质沈阳山楂圃保存的山楂种质资源为试材，首先利用高通量测序技术解析山楂属植物中ACLSV的全基因组序列；然后对感染ACLSV的山楂植株和健康植株进行转录组、小RNA组、降解组分析，明确ACLSV对山楂基因表达的影响，揭示源自ACLSV的病毒小干扰RNA（vsiRNA）组成和特性，预测山楂基因组中vsiRNA的潜在靶基因；最后通过转基因验证vsiRNA对山楂靶基因的转录后沉默作用及靶基因的生物学功能。本项目旨在从vsiRNA角度揭示ACLSV的致病机制，为ACLSV的防治奠定基础。

中文关键词： 山楂；病毒基因组；小RNA；基因表达

英文摘要： Apple chlorotic leaf spot virus (ACLSV) is one kind of RNA virus impairing the plants of Rosaceae family. ACLSV is one of the most economically important viral pathogens, but its pathogenic mechanism is unknown. Hawthorn (Crataegus spp.), originated in China, is not only one important economic forest tree, but also a good ornamental plant in Rosaceae family . In our previous study, we found that the hawthorn trees were infected by ACLSV. In this project, de novo assembly of the whole genome of ACLSV in hawthorn trees will be obtained by high-throughput sequencing. The effects of ACLSV on gene expression levels in hawthorn trees will be elucidated by transcriptome analysis. The structure and character of vsiRNAs originated from ACLSV will be carried out by high-throughput sequencing of small RNAs form the hawthorn plant infected with ACLSV. And then the target genes of vsiRNAs will be predicted by analysis the expression of genes of hawthorn with the complementary sequence of vsiRNA. The splitting site on predicted target gene will be validated by degradome sequencing. The effect of vsiRNA on target genes in hawthorn and functions of target genes will be validated by transferring the target gene and vsiRNA into plants. The aim of this project is to elucidate the pathogenic mechanism of ACLSV with vsiRNA. The results of this project will lay a foundation for ACLSV prevention.

英文关键词： Hawthorn;Virus genome;Small RNA;Gene expression