Introduction of the clusters of genes controlling pest and disease resistance from related crops to common wheat by means of the genetic genome rearrangement

通过基因组重排将控制相关作物抗病虫害的基因簇引入到普通小麦中

基本信息

批准号：
12306001
负责人：
ENDO Takashi
金额：
$ 29.96万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2003
项目状态：
已结题

项目摘要

The purpose of this study was to introduce small alien chromosome segments of rye and barley on which pest and disease resistance genes are located into common wheat by using the gametocidal system.1. Introduction of rye chromosomal segments into common wheatWe targeted rye 1R satellite on which are located four different resistance genes against rust and powdery mildew. We found five common wheat lines carrying the 1R satellite segments translocated onto common wheat chromosomes among the prescreened common wheat lines carrying aberrant 1R chromosomes. Using those lines, we tested the resistance against the rusts and the presence of the protein and DNA sequence of a seed storage protein secarin. We found two lines which possessed the resistance but lacked secarin Since we needed more breakpoints in the satellite, we screened more than 800 plants to obtain over 180 plants carrying aberrant 1R chromosome. Furthermore, we developed a universal PCR primer set which amplified any rye segments in order to screen for small rye segments in common wheat.2. Introduction of barley chromosomal segments into common wheatWe conducted a large scale screening for aberrations in barley chromosome 2H, 4H, and 7H by in situ hybridization. We studied the 2H aberration lines by HPLC (high performance liquid chromatography) to find that genes controlling the biosynthesis of an anti-fungal substance hordatine are located on the distal region of the short arm of chromosome 2H. We analyzed aberrant 4H and 7H chromosomes with various DNA markers ; especially, for chromosome 7H we conducted a kind of radiation hybrid mapping using ca. 80 7H aberration lines and different DNA markers (SSRs, AFLPs, -ESTs). We also developed a universal PCR primer set for barley chromatin.

本研究的目的是利用杀配子系统将带有抗病虫基因的黑麦和大麦外来染色体小片段引入普通小麦中。 1．将黑麦染色体片段引入普通小麦我们以黑麦1R卫星为目标，该卫星上有四种不同的锈病和白粉病抗性基因。我们在预筛选的携带异常 1R 染色体的普通小麦品系中发现了 5 个携带 1R 卫星片段的普通小麦品系易位到普通小麦染色体上。使用这些品系，我们测试了对锈病的抵抗力以及种子储存蛋白叶卡素的蛋白质和 DNA 序列的存在。我们发现了两个具有抗性但缺乏secarin的品系。由于卫星上需要更多的断点，我们筛选了800多株植物，获得了180多株携带异常1R染色体的植物。此外，我们还开发了一套通用PCR引物，可以扩增任何黑麦片段，以便筛选普通小麦中的小黑麦片段。2．将大麦染色体片段引入普通小麦我们通过原位杂交对大麦染色体2H、4H和7H畸变进行了大规模筛选。我们通过HPLC（高效液相色谱）研究了2H畸变线，发现控制抗真菌物质hordatine生物合成的基因位于2H染色体短臂的远端区域。我们用各种 DNA 标记分析了异常的 4H 和 7H 染色体；特别是，对于 7H 染色体，我们使用 ca 进行了一种辐射杂交作图。 80 个 7H 畸变线和不同的 DNA 标记（SSR、AFLP、-EST）。我们还开发了大麦染色质通用 PCR 引物组。