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利用已知植物抗病基因编码氨基酸保守区域NBS-LRR(核苷酸结合位点-富亮氨酸区域)设计了42个简并引物组合,运用抗病基因类似物多态性(resistance gene analog polymorphism,RGAP)分子标记技术,对中国春、中国春-长穗偃麦草双二倍体及其附加系和代换系基因组DNA进行PCR扩增。结果表明,共有38对引物组合获得扩增产物,其中35对在普通小麦中国春、中国春-长穗偃麦草双二倍体中能扩增出多态性,平均每个引物组合扩增出38.5个片段。在普通小麦背景下,共获得275条长穗偃麦草E基因组多态性谱带,占扩增总谱带数的17.44%,揭示出在普通小麦背景下E基因组和普通小麦A、B、D基因组间的高丰度遗传变异。另外,利用RGAP分子标记技术,构建了一套完整的长穗偃麦草1E~7E染色体的特异RGAP标记。为小麦背景中长穗偃麦草外源遗传物质的快速检测提供了新途径。
A total of 42 degenerate primer combinations were designed based on the amino acid conserved region NBS-LRR (nucleotide binding site-leucine-rich region) of known plant disease resistance genes. Resistance gene analogs Polymorphism and RGAP molecular markers were used to amplify the genomic DNA of the diploid and its ancillary lines and spring lines of Chinese Spring and Chinese Spring - The results showed that a total of 38 pairs of primers were used to amplify the amplified products. Among them, 35 pairs of primers amplified polymorphic DNA in the diploids of Chinese Spring and Chinese Spring - 38.5 clips. In the background of common wheat, a total of 275 E-genome polymorphism bands were obtained, accounting for 17.44% of the total number of bands amplified, revealing that the E genome and common wheat A, B, D High abundance genetic variation among genomes. In addition, a complete RGAP marker of chromosome 1E ~ 7E was constructed using RGAP molecular marker technology. This provided a new way for the rapid detection of foreign genetic material in Erigeroni elongatum in wheat background.