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

本研究的目的是利用杀配子系统将含有抗病和抗病基因的黑麦和大麦的外源小染色体片段导入普通小麦。将黑麦染色体片段导入普通小麦我们以黑麦1R卫星为目标，定位于4个不同的抗锈病和白粉病基因。在预筛选的携带异常1R染色体的普通小麦品系中，我们发现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。利用这些品系，我们测试了它们对锈病的抗性，以及种子贮藏蛋白secarin的蛋白质和DNA序列的存在。我们找到了两个具有抗性但缺乏铁蛋白的品系，因为我们需要在卫星上有更多的断裂点，我们对800多个植株进行了筛选，获得了180多个携带异常1R染色体的植株。此外，我们还开发了一套通用的PCR引物，可以扩增任何黑麦片段，以筛选普通小麦中的小黑麦片段。大麦染色体片段导入普通小麦我们用原位杂交的方法对大麦2H、4H和7H染色体的异常进行了大规模的筛选。我们通过高效液相色谱对2H突变系的研究发现，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。我们用不同的DNA标记对异常的4H和7H染色体进行了分析，特别是对7H染色体，我们用大约80个7H异常系和不同的DNA标记(SSR、AFLP、-EST)进行了一种辐射杂交定位。我们还开发了一套大麦染色质通用的聚合酶链式反应引物。

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.