Fe^+离子注入诱变莲花突变体的SRAP分析

SRAP ANALYSIS OF M_3 LOTUS MUTANTS INDUCED BY Fe^+ ION IMPLANTATION

为了在分子水平上对离子注入诱变莲花突变体进行鉴定,本研究对同一剂量Fe+离子注入诱变的白洋淀红莲(Baiyangdian red lotus)突变体及其对照的基因组进行SRAP扩增,并利用非变性聚丙烯酰胺凝胶电泳对SRAP-PCR扩增产物进行分析。在优化好的SRAP体系基础上,从121对引物组合中筛选出了10对可以稳定扩增出显著特异条带的引物,引物多态性为8.26%;将这10对引物的扩增产物用聚丙烯酰胺凝胶电泳检测,共扩增出了215条带,多态性条带为83条,多态率为38.6%;突变体1扩增出141条带,和对照相比,特异条带共22条,条带变异率为15.6%,而突变体2～6的条带的变异率分别为16.4%、17.1%、16.9%、18.2%和20.5%。表明Fe+离子注入莲花种子可导致其基因组DNA发生变异,同一剂量Fe+离子注入诱变具有随机性。

To examine and determine the lotus mutants induced by the same Fe^＋ ion implantation at the molecular level,the SRAP technique was used,and the non denatured polyacrylamide gel electrophoresis was undertaken to analyze the PCR products. At the optimized SRAP reaction condition,of the 121 primer pairs tested,10 primer pairs could amplify stable and remarkable specific bands,with primer polymorphism of 8.26%. This 10 primer pairs amplified a total of 215 bands,83 of which were polymorphic,and the percent of polymorphic bands was 38.6%. A total of 141 bands were amplified for the mutant 1,22 of which were different from the control with the variation ratio of 15.6%. However,the variation ratios of mutant 2 to 6 were 16.4%,17.1%,16.9%,18.2% and 20.5% respectively. The results indicated that Fe^＋ ion implantation into the seeds of Baiyangdian red lotus could induce random genetic DNA variations.