论文部分内容阅读
小麦近缘种簇毛麦携带许多尚未克隆的抗病(R)基因。NBS-LRR类型的R基因占已克隆植物R基因的绝大多数,因此,本研究根据NBS-LRR类型R基因的保守序列设计引物,从簇毛麦基因组DNA和cDNA中扩增获得23条相关序列。基于其中5条抗病基因同源序列(RGAs)H-56/d6、H-66/b2和CDS40设计引物,对小麦、簇毛麦、硬-簇双二倍体及其杂种以及已知携带个别簇毛麦染色体或染色体臂的小麦材料进一步进行PCR扩增,结果表明:3对引物均可对簇毛麦、硬-簇双二倍体进行特异扩增;同时,源于序列H-66/b2的引物可对1VL和6VL染色体臂进行特异扩增;源于序列CDS40的引物可在同时携带1VL和2VS或同时携带2VS和4V的小麦材料以及具有6VL的小麦材料中特异扩增,而H-56/d6的引物在携带1VL、2VS、4V和6V染色体臂或染色体的小麦背景中都不能获得目的片段的扩增。这些结果不仅为外源染色体臂在小麦背景中的追踪与鉴定提供了新的分子标记,而且这些标记还与外源染色体或染色体臂上的抗病基因或抗病基因同源物紧密连锁或共分离。
The wheat-related species Rafflesieae host many un-cloned disease resistance (R) genes. NBS-LRR type R gene accounted for the vast majority of cloned plant R genes, therefore, in this study, based on the NBS-LRR type R gene conserved sequence design primers, genomic DNA and cDNA amplified from Hamster 23 related sequence. Based on the primers designed for five resistance gene homologous sequences (RGAs) H-56 / d6, H-66 / b2 and CDS40, wheat, Hamster, hard-cluster diploid and their hybrids, The results showed that three pairs of primers could specifically amplify the diploid of hardwood-cluster and double-diploid of hardwood-cluster, respectively. At the same time, the wheat from the sequence H-66 / b2 primers specifically amplify the 1VL and 6VL chromosomal arms; primers derived from the sequence CDS40 can be specifically amplified in 1VL and 2VS carrying either 2VS and 4V of wheat material and 6VL of wheat material carrying both Primers for H-56 / d6 did not achieve amplification of the target fragment in wheat backgrounds carrying 1VL, 2VS, 4V and 6V chromosomal arms or chromosomes. These results not only provide new molecular markers for tracing and identification of exogenous chromosome arms in the wheat background but also are closely linked to or contiguous with disease-resistance genes or disease-resistance gene homologs on exogenous chromosomes or chromosomal arms Separated.