离子注入介导麻黄和甘草总DNA转化苜蓿的初步研究

Ion Implantation Mediated Ephedra and Glycyrrhiza Genome DNA Transformation into Medicago sativa

目的:探索离子注入介导麻黄和甘草总DNA转化苜蓿的方法与效果。方法:氩离子(Ar+)注入介导麻黄和甘草总DNA在2个品种的苜蓿中转化,对种子发育正常的T1代苜蓿单株进行生物碱类、黄酮类和皂苷类物质进行定性检识。结果:苜蓿种子经离子注入介导转基因处理后,T1代结荚率很低,复合DNA处理的183和283的结荚率分别为6.40%和8.33%,正常发育的种子分别为2.40%和5.83%。获得了种子发育良好的苜蓿T1代单株50株,其中6个单株根或茎的天然产物定性检识呈阳性。结论:Ar+注入介导外源DNA大分子的转化对苜蓿种子的发育具有很大影响,获得的转基因苜蓿T1代种子,为进一步开展相关化学证据和分子证据的研究提供了宝贵的材料。

Objective： In order to investigate the methods and effects that the total DNA from Ephedra and Glycyrrhiza were transformed into alfalfa by ion implantation. Methods： Ephedra and Glycyrrhiza total DNA were transfonned into two kinds of alfalfa by Ar^＋ implantation. The alkaloids, flavonoids, saponins were tested qualitatively to the well- developed seeds of T1 generation alfalfa. Results： The seed- setting rate of T1 generation alfalfa were 6.40% and 8.33% with complex DNA treatment 183 and 283, respectively, and normal seed development was only 2.40% and 5.83%. 50 strains of T1 generation alfalfa with well- developed seeds were obtained, among which the natural products of 6 strains＇ roots or stalks were showed positive reaction by qualitative analysis. Conclusion： The great influence on the normal development of alfalfa seeds were induced by Ar^＋ impLantation mediated the exogenous DNA transformation. The obtained seeds of T1 generation alfalfa were provided the valuable materials for the further research on the chemical and molecular evidences.