葡萄无核基因的SCAR标记及Southern blot分析

SCAR marker linked to seedless genes in grapes and southern blot analysis

根据无核基因特异RAPD标记的序列进行分析,合成1对特异引物P1+P3(序列分别是P1:5′CTCATCTTCTTGATGGTGAT3′;P3:5′AAGTGAAGAACATCATTAAGAAC3′),对30个葡萄材料进行PCR扩增,成功地将RAPD标记转换成SCAR标记。并对含有该标记序列的重组质粒进行Hind 和Spe 双酶切,获得的310bp片段回收后制成探测葡萄无核基因存在与否的杂交探针,成功地对7个葡萄品种进行Southernblot分析,实现了分子标记检测葡萄无核基因的目的。

This paper reported a rapid detection of genetic marker linked to seedless genes in 30 grape varities using SCAR marker and Southern blot analysis.One pair of specific primers were designed based on the sequence analysis of the RAPD marker linked to seedless genes in grapes,and synthesized to perform PCR.The RAPD marker was transformed into SCAR marker successfully.Two restriction enzymes were selected to digest recombinant plasmids and the genome DNA of seven seeded and seedless grape cultivars.The 310 bp specific fragment obtained from recombinant plasmids was made into a probe after DIG labling to detect the seedless genes in grapes.The digested genome DNA showed the existence of the seedless genes in seedless cultivars using southern blot,whereas the seeded ones were not.