利用SRAP标记快速检测西瓜杂交种子纯度

Using SRAP Markers for Rapid Detection of Zaojia( 84-24) Watermelon Hybrid Seed Purity

【目的】以设施、露地兼用西瓜杂交品种早佳(84-24)及其父本、母本为试验材料,利用SRAP分子标记技术进行DNA指纹图谱构建,筛选出能够快速检测西瓜早佳(84-24)杂交种子纯度的SRAP标记。【方法】选取28对SRAP引物,对供试西瓜品种早佳(84-24)及其父、母本的基因组DNA进行扩增,筛选出扩增多态性好、条带清晰、稳定的SRAP引物作为候选标记,利用100粒杂交种子进行纯度检测准确性验证。【结果】28对供试引物中有22对引物在F1及其父、母本之间扩增出多态性条带,多态性比率为78.6%,其中有1对为父本显示,母本缺失的显性SRAP引物(Me6F/Em2R)。用显性引物(Me6F/Em2R)对100粒西瓜杂交种子进行纯度检测,经聚丙烯酰胺凝胶电泳分析,纯度为99.0%,与田间形态学鉴定结果(99.1%)基本一致。【结论】利用SRAP标记(Me6F/Em2R),结合聚丙烯酰胺凝胶电泳检测,可以对西瓜杂交种早佳(84-24)的纯度进行快速、准确鉴定。SRAP分子标记技术在西瓜杂交种子纯度室内快速检测中有很大的应用前景。

【Objective】To screen out one SRAP marker to detect seed purity,the watermelon hybrids of Zaojia（ 84-24） and its paternal and female lines were used to study the polymorphorrism of amplifiable spectral bands.【Method】Polymorphic primer pairs were selected from 28 SRAP primer pairs for Zaojia（ 84-24）hybrids and their parents in this experiment. The SRAP primers with good polymorphism,clear bands and stable primers were selected as candidate markers,and the purity of the 100 hybrid seeds was tested and verified.【Result】The results showed that 28 pairs of primers had 22 pairs of primers,and polymorphic bands were amplified between F1 and its parents. The ratio of polymorphism was 78. 6%,among which,there was only one pair of complementary type SRAP primer（ Me6 F/Em2 R） with male display and lack of female parent,The result showed the purity of hybrid seeds was 99. 0%,which was basically the same as the field morphological identification（ 99. 1%）.【Conclusion】SRAP primer（ Me6 F/Em2 R）,in combination with Polyacrylamide gel electrophoresis can be used to detect the purity of Zaojia（ 84-24） hybrid seed rapidly and accurately. There is a great prospect of application in using this method.