项目名称： 移虫时间对西方蜜蜂蜂王生殖发育的影响及分子机理的研究

项目编号： No.31502020

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 畜牧学与草地科学

项目作者： 殷玲

作者单位： 江苏农牧科技职业学院

项目金额： 20万元

中文摘要： 繁殖性能是蜂王育种工作中关注的重点，移虫时间对蜂王生殖发育影响显著。本研究对孵化后0h、24h、48h、72h移虫至王台内发育的4日龄幼虫卵巢形态、保幼激素及蜕皮激素进行检测，并对各组出房蜂王进行繁殖性能测定。利用RNA-Seq技术对各组4日龄幼虫卵巢组织进行转录组差异表达分析，对差异表达基因进行GO功能及Pathway显著富集分析，探究影响蜂王幼虫早期生殖发育的功能基因及生物学通路。运用Bisulfite-seq 技术对各组4日龄幼虫卵巢组织全基因组5mC信息扫描和差异性甲基化区域分析，探明影响蜂王幼虫早期生殖发育的表观遗传学机制。整合卵巢组织转录组及全基因组甲基化数据，结合各组蜂王繁殖性能分析，运用RT-PCR及western blot 技术验证，筛选调控蜂王生殖发育的关键基因及调控网络。本研究为指导蜜蜂育种工作提供理论基础，同时为丰富和完善蜜蜂级型分化研究提供新的数据和理论依据。

中文关键词： 西方蜜蜂；蜂王；生殖发育；转录组；DNA甲基化

英文摘要： Improving reproductive performance is an important goal of honeybee breeding, and reproductive development of the queen is greatly affected by the age of transferring larvae. In this study, ovaries morphology, JH and Ecd of 4th instars transferring in 0h, 24h, 48h, 72h are detected, and the reproductive performance of queens developed from different transferring age are monitored. The transcriptomes of 4 days old larvaes with different transferring age are analyzed using RNA-Seq method, and effective genes and regulatory pathways effected reproductive development of queen are screened through GO and Pathway enrichment analysis. The whole genome DNA methylation level of 4 days old larvaes with different transferring age are analyzed using bisulfite sequencing method, the enigenetic mechanism for reproductive development of queen are surveyed through differential methylation regional analysis. The data of transcriptome and genome-wide methylation are integrated, and the reproductive performance analysis of 4th instars and queens developed from different transferring age is taken into account to screened for effective genes and regulatory pathways. RT-PCR and western blot technology is used to validate the the genes effected reproductive development of queen and the regulatory network. This study provide theoretical basis for guiding the bee breeding work, and provide new research data and theoretical basis to enrich the study on honey bee caste determination.

英文关键词： Apis mellifera;queen;reproductive development;transcriptome;DNA methylation