SSR与SRAP分子标记在烤后烟叶中的应用及烤烟遗传多样性分析

Application of SSR and SRAP molecular markers in cured tobacco leaves and genetic diversity analysis of flue-cured tobacco varieties

为筛选出适合烤后烟叶的分子标记并用于烤烟遗传多样性分析,以烤后烟叶为材料,利用毛细管电泳技术检测SRAP与SSR标记在烤后烟叶中的多态性和稳定性,并基于稳定性较好的SSR分子标记分析了云烟87、云烟97、云烟116、翠碧1号、红花大金元、中烟100、K326、KRK26、NC297等9个烤烟品种的遗传多样性。结果表明:与新鲜烟叶相比,烤后烟叶DNA降解较为严重,其降解主要发生在烘烤过程中干筋期的中后期。与SRAP相比,SSR标记在烤后烟叶中扩增稳定性更好,但多态性略差。此外,NTGS66290、NTGS66292、NTGS66295、PT30008、PT30169、PT30361、PT30403和PT30424等8对SSR引物能将9个烤烟品种进行有效区分,其遗传相似系数为0.250~0.900,平均值为0.622。

In order to screen out molecular markers suitable for the genetic diversity of cured tobacco leaves,the polymorphism and stability of SRAP and SSR markers in cured tobacco leaves were detected by capillary electrophoresis.The genetic diversities of 9 flue-cured tobacco cultivars,including Yunyan 87,Yunyan 97,Yunyan 116,Cuibi 1,Honghuadajinyuan,Zhongyan 100,K326,KRK26 and NC297,were analyzed based on SSR marker with good stability.The results showed that the DNA degradation of cured tobacco leaves was more serious than fresh tobacco leaves,and the degradation mainly occurred in the middle and late periods of stem-drying stage.Compared with SRAP markers,SSR markers had better amplification stability in cured tobacco leaves,while their polymorphisms were slightly worse.In addition,eight pairs of SSR primers such as NTGS66290,NTGS66292,NTGS66295,PT30008,PT30169,PT30361,PT30403 and PT30424 can effectively distinguish 9 flue-cured tobacco varieties.The genetic similarity coefficients were from 0.250 to 0.900 with the average value of 0.622.