权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Molecular analysis of Kr1 gene responsible for crossability of common wheat with alien species

负责普通小麦与外来物种杂交的 Kr1 基因的分子分析

基本信息

批准号：
12660003
负责人：
KOBA Takato
金额：
$ 2.3万
依托单位：
CHIBA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

Common wheat is one of the most important crops as well as rice and maize. For the purpose of improvement of wheat cultivars, alien genes of the genus rye and Aegilops have been incorporated into common wheat, and many cultivars having cold resistance, desease and insect resistance, and high quality proteins have been produced. However, in the case of introduction of alien genes, hybridization between wheat and alien species have been a big problem. Crossability of common wheat with rye have been clarified to be located on the chromosomes of homoelogous group 5 of common wheat. The effects of Kr genes are expressed on the stigma and ovule, and inhibit the penetration and elongation of pollen tubes. In the present research, on the purpose of clarifying the mechanism of crossability, we tried to isolate the DNA fragments which are correspond to the Kr1 gene, and clone the fragments and identify the nucleotide sequences. Materials used are 20 recombinant inbred lines produces from the cro … More ssing between Chinese Spring wheat (kr1), which has high crossability with rye, and its 5B chromosome substitution line derived from cv.Mara (Kr1) which has low crossability. These lines were classified according to their crossability with rye. In the flowering period, pistils of wheat lines were collected and classified in to two groups and their bulked mRNAs were isolated. Through reverse transcription method, cDNAs were synthesized and polymorphism were detected by DD-RAPD and DD-AFLP method. The results from DD-RAPD has a problem on the repeatability and not reliable. In the case of DD-AFLP, after the analysis using 1723 primer pairs, 62 specific cDNA fragments for Kr1 lines were identified. Among them, 31 fragments were excised from the gel, and 24 clones were obtained. Nucleotide sequences of them showed that they were homologous to the genes for the protein of two factors control system of Arabidopsis thaliana and the precursor of cystein protenase. In the future, northern analysis will be necessary to examine the tissue specific gene expression in pistils of common wheat. Less

普通小麦是最重要的作物之一，还有水稻和玉米。为了改良小麦品种，将黑麦属和黑麦属的外源基因引入普通小麦中，培育出了许多具有抗寒、抗病、抗虫和优质蛋白质的小麦品种。然而，在引进外来基因的情况下，小麦与外来种的杂交一直是一个大问题。普通小麦与黑麦的杂交能力确定在普通小麦同系第5群的染色体上。Kr基因在柱头和胚珠上表达，抑制花粉管的穿透和伸长。在本研究中，为了阐明杂交性的机制，我们试图分离Kr1基因对应的DNA片段，克隆片段并鉴定核苷酸序列。材料为与黑麦杂交能力强的中国春小麦（kr1）及其5B染色体代换系（cv）杂交产生的20个重组自交系。杂交性低的Mara （Kr1）。这些系根据与黑麦的亲和性进行分类。在花期收集小麦品系的雌蕊，将其分为两组，并分离出雌蕊的大块mrna。通过逆转录法合成cdna，并通过DD-RAPD和DD-AFLP法检测多态性。DD-RAPD的结果存在重复性差、可靠性差的问题。在DD-AFLP中，使用1723对引物分析后，鉴定出62个Kr1系特异性cDNA片段。其中，从凝胶中切除31个片段，获得24个克隆。核苷酸序列表明，它们与拟南芥双因子控制系统蛋白基因和半胱氨酸蛋白酶前体基因同源。今后，对普通小麦雌蕊组织特异性基因表达的研究将是必要的。少