RAPD及SRAP 2种标记对苦瓜‘如玉5号’杂交种纯度的鉴定

Purity Identification of Hybrid Bitter Gourd ‘Ruyu No.5’ Using RAPD and SRAP Markers

为获得理想的方法鉴定‘如玉5号’苦瓜杂交种子的纯度,以苦瓜杂交一代品种‘如玉5号’及其父母本为材料,利用RAPD(随机扩增多态性DNA)及SRAP(相关序列扩增多态性)2种技术进行其基因组总DNA指纹分析,以获得F1代杂交种与其亲本差异目的基因条带。经过对3种苦瓜新鲜幼嫩叶片总DNA提取,RAPD-PCR和SRAP-PCR扩增以及扩增产物的琼脂糖凝胶电泳分析,在供试的52条RAPD引物及144对SRAP引物中,共筛选出2对SRAP引物(Me2-Em9、Me3-Em8)能区分杂交种与其母本种子,1条RAPD引物(R10)能区分杂交种与其父本种子,通过SRAP(引物Me3-Em8)及RAPD(引物R10)2种分子标记技术对来自于河西走廊3地‘如玉5号’苦瓜种子进行检测,结合田间观察,结果表明,该方法成本低,操作简单,能很好地对‘如玉5号’苦瓜杂交种子的纯度鉴定。

Using RAPD（random amplified polymorphic DNA） and SRAP（sequence- related amplifiedpolymorphism） molecular markers technology, the DNA fingerprints of‘Ruyu No.5＇bitter gourd and itsparents were analyzed to obtain the different genes differentially expressed in order to identify the purity ofhybrid seeds eventually. After repeated experiments, only 1 RAPD primer（R10） screened out from 52 couldamplify the characteristic polymorphic bands of F1 generation with its male parent and 2 SRAP primers（Me2-Em9, Me3-Em8） screened out from 144 could amplify the characteristic polymorphic bands of F1 generationwith its female parent. With the two markers from RADP（R10） and SRAP（Me2-Em9） possessing high stabilityand repeatability, the seed of‘Ruyu No.5＇from Hexi Corridor in Gansu Province were identified, and theresult was roughly according to the identification from field. The result showed that this method for low cost andeasy operation can be perfect to identify the purity of‘Ruyu No.5＇bitter gourd seeds.