为了建立草莓品种指纹图谱数据库,从而对不同草莓品种进行分子鉴定,以早光、因都卡、达赛莱克特、森加拉等38个欧美草莓栽培品种为试材,利用8%非变性聚丙烯酰胺凝胶电泳进行扩增产物检测。通过对各种影响因素不同浓度梯度的比较,得出优化的SSR扩增反应体系,25μL PCR的反应体系中各成分为：1×Buffer;2.5 mmol.L-1MgCl2;0.4μmol.L-1引物;1.5 U Taq酶;0.2 mmol.L-1dNTP;60 ng DNA。从44对引物中筛选出带型清晰、多态性丰富的10条SSR引物。最终确定了4对SSR引物作为核心引物对38个草莓品种进行了分子图谱的构建,29个草莓品种可以完全与其他品种区分开。
Establishment of SSR fingerprinting database for 38 cultivars of straw- berry （Fragaria ananassa） native to Europe and America
The total DNA of 38 strawberry cultivars originating in Europe and America , such as Earliglow, Induka, Darselect ,Senga, Litessa, were amplified by 44 pairs of SSR primers. The PCR products were separated in 8% polyacrylamide sequencing gels. Some factors which affected amplified products, such as dNTPs, Taq DNA polymerase,primer and so on were also studied. Optimum condition was as follows：25 μL PCR reaction system consisted of 1 xPCR Buffer, 2.5 mmol ·L^-1 MgCl2, 0.4 μmol primer, 0.2 mmol dNTP, 1.5 U Taq polymerase and 60 ng DNA. The amplification conditions were 94 ℃ for 1 rain, 35 cy- cles of 94 ℃ for 30 s,annealing temperature for 30 s ,72 ℃ for 30 s and a final extension step at 72 ℃ for 7 rain. Ten pairs of primers which could amplify stable and distinct band were selected. The electrophoresis results of the 38 strawberry DNA samples were evaluated and analyzed. Eventually, 29 cuhivars could be distinguished by the fingerprint map constructed by only 4 pairs of SSR primers.
国家科技期刊平台
NSTL
万方数据
维普
