桃果实非酸/酸性状SCAR标记的转化与验证

 根据筛选到的与桃果实非酸/酸性状紧密连锁的3个AFLP标记特异片段序列信息, 设计特异引物BFPA1/A2、BFPB1/B2和BFPC1/C2。3个特异引物在‘京玉’桃、‘美味’油桃及其正反交F¹代69株群体中的PCR检测结果表明: 引物BFPB1/B2在果实性状表现为酸的后代个体中扩增出分子量大小为158 bp的特异条带, 且根据特异片段AT-CTA₁₉₉不同部位设计的引物都能稳定扩增, 只是片段大小有所差异;特异扩增检测F¹群体分离结果与原先AFLP_AT/CTA标记完全一致, 表明AFLP标记已成功转化为SCAR标记,该SCAR BFPB1/B2标记与D基因的连锁距离为2199 cM。利用SCAR BFPB1/B2标记对‘大久保’桃ב兴津’油桃的F²群体60株个体的果实非酸/酸性状进行检测, 基因型与表型性状符合率为96.7% , 并且表现为共显性标记。特异引物BFPA1/A2和BFPC1/C2的扩增多态性条带在亲本及后代中消失。

Development and Validation of SCAR Markers Linked to the Non-acid/acid Trait of Fruit in Peach

In this paper, specific primers of BFPA1/A2 , BFPB1/B2 , BFPC1/C2 were designed respectively according to the sequence of three specific fragments from AFLP markerswhich linked to non-acid/acid trait of peach. A 158 bp polymorphic band was amplified from the DNA of 69 F¹ population of the intra-specific cross between cultivars‘Jingyu’and‘Flavortop’with the primer pair BFPB1/B2. Furthermore, the steady and specific amp lification can be got with the primers designed in different location of the AFLP fragmentAT-CTA199. The SCAR marker had the same segregation pattern as the original AFLP_AT/CTA marker with a linkage distance of 2199 cM and the AFLP marker was converted into SCAR marker successfully. The SCAR_BFPB1/B2 marker was tested in 60 F² p rogeny of‘Okubo’and‘Xingjin’nectarine, and the results showed that the ac2 cordance ratio of genotype and phenotype was 96.7%. The SCAR marker was a codominant marker and could distinguish the heterozygous and homozygous. Polymorphism disappeared in parents and progeny with specific primers BFPA1 /A2 and BFPC1 /C2.