RAPD分析氮离子注入甜菊种子后的幼苗基因组DNA变异

RAPD ANALYSIS OF SEEDLING GENOMIC DNA VARIATIONINDUCED BY N^+ IONS IMPLANTATION IN STEVIA SEEDS

应用ＲＡＰＤ 技术检测经低能氮离子注入甜菊纯系种子引起的幼苗基因组ＤＮＡ 变异。筛选出ＯＰＪ系列中的１５ 种引物对实验及对照基因组ＤＮＡ 进行了ＰＣＲ 扩增，共获扩增片段１０３ 条，分子量在０．３ － ３ｋｂ 之间，其中５ 种引物ＯＰＪ－ １ ，７ ，９，１１ ，１２ 扩增出差异片段１２ 条。结果表明，低能氮离子注入甜菊种子可引起体内基因组ＤＮＡ 发生突变；ＲＡＰＤ 技术是检测基因组ＤＮＡ 发生诱变的一种简便、有效方法。本文同时探讨了离子强度和Ｔａｇ ＤＮＡ 聚合酶用量对甜菊ＲＡＰＤ 分析结果的影响，以及氮离子注入诱变效应的可能机制。

The paper reports the genomic DNA variation induced by N+ ion implantation in Stevia seeds using RAPD analysis. The total of 103 bands has been amplified from Stevia genomic DNA with fifteen screened primers. The molecular weight of every amplified fragment is between 0.3kb and 3kb. There are twelve polymorphic bands have been amplified by four 10bp primers (OPJ-1, 7,9,11,12). The result show that implantation of N+ ion beam can induce genomic DNA variation. RAPD analysis is a simple and efficient method in detecting genomic DNA variation. The effect of different ion intensity and the quantity of Tag DNA polymerase on the result of RAPD analysis as well as the possible mechanism of biological effects induced by N+ ions implantation is also discussed.