项目名称: 普通小麦籽粒过氧化物酶活性基因克隆、表达机理分析及功能标记开发
项目编号: No.31260327
项目类型: 地区科学基金项目
立项/批准年度: 2013
项目学科: 农业科学
项目作者: 耿洪伟
作者单位: 新疆农业大学
项目金额: 53万元
中文摘要: 面粉颜色是决定小麦品质的重要指标,对面条、馒头等面制品品质有重要影响。籽粒过氧化物酶(Peroxidase,POD)活性是影响面粉和面制品颜色的重要因素。本研究拟通过选用Sunstate/周麦18 F6重组自交系群体和250份中国冬小麦主栽品种,进行POD活性的QTL分析;应用同源克隆结合PCR验证的方法,克隆与颜色性状紧密相关的POD基因并分析其在不同品种中的等位变异,根据各等位基因之间的序列差异开发基因的功能标记,并分析基因型与表型之间的关系;利用实时定量PCR和Western Boltting技术分析籽粒不同发育时期POD位点不同等位基因的表达模式,利用TILLING技术验证小麦过氧化物酶活性基因的功能。研究结果将为分子标记辅助选择快速改良过氧化物酶活性提供有效的功能标记,揭示POD基因的分子机理,最终为开展小麦过氧化物酶活性基因分子生物学研究和面粉颜色改良提供科学依据。
中文关键词: 小麦;过氧化物酶;基因克隆;全基因组关联分析;功能标记
英文摘要: Flour color is an important trait in the assessment of flour quality and exerts significant influence on the qualities of Chinese noodles, steamed bread and other products. Peroxidase (POD) activity in grain is an important factor determining the color of ?our and end-use products of wheat. In the present study, an F6 recombinant inbred line (RIL) population derived from the Sunstate/Zhoumai 18 cross will be used to map QTLs for POD activity. The POD genes will be cloned by the method of in silico cloning in combination with PCR amplification, and the functional markers will be developed for the genes according to their allelic varants among different wheat cultivars, which will be validated with the RIL population and 250 Chinese winter wheat cultivars. The expressions of POD genes during grain development will be analyzed by real-time PCR (RT-PCR) and Western Blotting, and the gene function will be studied by TILLING. This study will provide useful information about the molecular mechanism of POD gene expression and its association with wheat quality, and functional markers for the improvement of POD activities in wheat breeding.
英文关键词: Wheat;Peioxidase;Gene cloning;GWAS;Functional marker