石榴叶片SRAP体系优化及其在白花芽变鉴定中的应用

Optimization of SRAP System of Pomegranate(Punica granatum L.)and Its Application on Identification of a White Flower Mutation

采用优化的SRAP-PCR反应体系,对红花石榴母株上的白花变异枝进行鉴定和分析.结果表明,适宜石榴的SRAP-PCR反应体系中含dNTP 0.2 mmol/L、Mg2+2 mmol/L、引物0.4μmol/L、DNA模板0.8 mg/L、Taq聚合酶50 000 U/L.600对SRAP随机引物组合中有580对引物组合有较好的扩增效果,其中Me30/Em7引物组合在母株和变异枝条间扩增出1条长度为78 bp的稳定差异条带.根据该差异片段序列设计的特异引物在母株中有68 bp扩增条带,在变异枝条中没有扩增条带.表明白花变异枝条的产生可能与母株DNA片段缺失有关.

Molecular identification and analysis were conducted on the white flower variation of the red flower stock plant of pomegranate using an optimized SRAP-PCR system.The results showed that the optimum system contained dNTPs 0.2 mmol/L,Mg2＋ 2 mmol/L,primer 0.4 μmol/L,template DNA 0.8 mg/L and Taq polymerase 50 000 U/L.Five hundred and eighty out of six hundred primer combinations had more amplified bands,in which a stable 78 bp differential band amplified by Me30/Em7 combination was found between the stock plant and the variation.A 68 bp specific band was amplified in the red flower genotype but not in the variation using one specific primer pair designed according to the sequence.The evidence suggested that the variation was a sport,which might be caused from the DNA fragment deletion of the stock plant.