项目名称： 影响槟榔江水牛乳脂性状重要功能SNP的鉴定

项目编号： No.31460582

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 苗永旺

作者单位： 云南农业大学

项目金额： 52万元

中文摘要： 水牛奶乳脂含量高，在乳脂组成上特征明显，但水牛乳脂性状的遗传机制尚不清楚。槟榔江水牛是近年在云南发现的我国第一个本土河流型水牛，具有较高的种用价值。本项目拟以该水牛繁育核心群为研究对象，按照乳中乳脂含量高低，将其分成高乳脂和低乳脂含量组，从泌乳的重要器官--乳腺入手，针对水牛乳脂性状的分子遗传基础开展研究。先采用RNA-Seq技术对上述两组水牛3个不同泌乳时期的乳腺组织样品池进行转录组测序分析，筛查两组水牛间的差异表达基因，从中选出影响乳脂性状的功能候选基因，并通过RNAi和基因过表达技术对其功能进行体外验证，确定影响水牛乳脂性状的重要功能基因；进一步通过对功能基因SNP的筛选及SNP位点与乳脂性状的大样本关联分析、EMSA实验，鉴定出影响水牛乳脂性状的功能SNPs。最终阐明影响水牛乳脂代谢的关键基因和调控网络，从分子水平上揭示水牛高乳脂特性的遗传基础，为奶用水牛乳脂性状的选育提供依据。

中文关键词： 槟榔江水牛；乳脂性状；功能基因；转录组测序；功能SNP的鉴定

英文摘要： Water buffalo milk contains a high level of milk fat content and the characteristics of buffalo milk fat are obvious in composition. However, the genetic mechanism of buffalo milk fat trait is unclear. Binglangjiang buffalo has high value in dairy breeding, which is the first indigenous river buffalo found in Yunnan province of China in recent years. This project aims to dissect the molecular genetic basis of buffalo milk fat trait from the mammary gland which is the important organ of milk secretion by taking the purebred breeding core herds of Binglangjiang buffalo as the research object, which will be divided into two groups, namely, the higher milk fat group and the lower milk fat group according to the level of milk fat content. Firstly, we will performed a transcriptome sequencing and the corresponding data analysis on the pooled breast tissue samples from three different stages of lactation (15, 90 and 250 d relative to parturition) of the previous two groups, then screening differentially expressed genes between the two groups from which choose the important functional candidate genes related to milk fat trait. To determine the functional genes affecting milk fat yield and composition, we will further verify the function of candidate genes in vitro by RNA interference (RNAi) and gene over-expression technology. Finally, we intend to identify the functional SNPs which influence the milk fat trait of water buffalo over the SNPs screening of functional genes, EMSA experiment detection and the association analysis of large sample data between the genotypes of the SNP loci and the milk fat trait. This project will ultimately unravel the key genes and their regulatory networks related to milk fat metabolism in water buffalo, reveal its molecular genetic basis of high milk fat performance and will provide a basis for milk fat trait breeding of dairy buffalos.

英文关键词： Binglangjiang buffalo;milk fat trait;functional genes;RNA-Seq;identification of functional SNPs