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用260个10bp随机引物对中国春（CS）、ph1b突变体及其F2分离群体基因组DNA进行RAPD分析，筛选出两个特异的RAPD标记OPR7936和OPR17524。经Southern分析，并结合减数分裂MI染色体配对分析，认为这两个RAPD标记位于5BL染色体ph1b基因缺失区中，可以作为ph1b基因的分子标记。经过连锁分析，测得OPR7936与Ph1基因之间的距离为11．4cM，OPR17524与Ph1基因之间的距离为5．0cM，均位于Ph1基因的远着丝粒端。测序后，经同源性比较发现这两个DNA序列可能是Ph1b基因区中的两个不同位点上的新序列，并有可能成为Ph1基因区图谱新的探针。根据这两个PAPD克隆片段的两端序列，分别设计出一对24bp的SCAR引物，可特异扩增Ph1基因的一条带（SCR7823和SCR17403），而ph1b基因缺少各自的特征带，其扩增产物无需电泳分离，只需在反应管中加入溴化乙锭，紫外灯下直接根据荧光反应即可鉴定出ph1b基因型，为今后ph1b基因的转育进行标记辅助选择又提供了两个易于操作、有效的分子标记。
Two hundred and twenty random primers were used for RAPD analysis of genomic DNA of Chinese spring (CS), ph1b mutant and F2 segregating population. Two specific RAPD markers, OPR7936 and OPR17524, were screened out. Southern analysis combined with MI chromosome pairing analysis suggested that these two RAPD markers were located in the deletion region of ph1b gene on chromosome 5BL and could be used as molecular marker for ph1b gene. After linkage analysis, we found that the distance between OPR7936 and Ph1 was 11.4cM and the distance between OPR17524 and Ph1 was 5.0cM, all located at the distal centromere of Ph1 gene. After sequencing, it was found by comparison of homology that these two DNA sequences might be new sequences at two different sites in Ph1b gene region and might become new probe of Ph1 gene region map. According to the two end sequences of these two PAPD clones, a pair of 24 bp SCAR primers were designed respectively to amplify one band of Ph1 gene (SCR7823 and SCR17403), while ph1b gene lacked its own band, and its amplification product Without electrophoresis separation, just add ethidium bromide in the reaction tube, and the ph1b genotype can be identified directly by the fluorescence reaction under the UV light, which provides two convenient operations for the future selection and selection of the ph1b gene transfer, Effective molecular markers.