秦美猕猴桃高频遗传转化再生体系的建立

Study on Regeneration System of High Frequency Genetic Transformation Kiwifruit(Qinmei)

以秦美猕猴桃茎段为外植体,在原代培养成功获得无菌试管苗的基础上,进行愈伤组织诱导培养。将获得的秦美猕猴桃愈伤组织转入再分化培养基中,采用正交设计试验,添加不同种类及不同浓度的植物生长调节物质,组成9种不同的再生植株分化培养基,对秦美猕猴桃愈伤组织进行再分化培养。结果表明,诱导培养基中添加1.0 mg/L 6-BA+0.1 mg/L NAA暗培养,有利于秦美猕猴桃愈伤组织的形成,诱导率达100.0%;再分化培养基中添加3.0 mg/L 6-BA+0.1 mg/L NAA+50 g/L蔗糖,能明显提高秦美猕猴桃愈伤组织再分化率。这种方法把诱导愈伤组织与再分化分开进行,可以获得更多的愈伤组织进行再分化培养基的优化,也有利于进行秦美猕猴桃的遗传转化研究。

The stems of Actinidia deliciosa cv.Qinmei were used as explants to induce callus after successfully gained aseptic seedling in primary culture.The obtained calli were then placed on regeneration medium.Different concentration of different plant growth regulators such as 6-BA and NAA was added to compose the 9 different plant regeneration medium of L9（34） orthogonal design.The results showed that the induction medium adding 1.0 mg/L 6-BA and 0.1 mg/L NAA in dark culture condition was beneficial to callus induction,on which the inducing rate could be as high as 100%.The suitable regeneration medium was MS＋ 3.0 mg/L 6BA＋0.2 mg/L NAA＋50 g/L sucrose;it could obviously improve the frequency of regenerated shoots.The callus induction and shoot regeneration was separated by this method;and more callus could be obtained for further study;thus was advantageous to kiwifruit genetic transformation.