利用农杆菌侵染花柱头法进行海岛棉遗传转化

目的]探索一种更为高效、简便的海岛棉遗传转化技术,为海岛棉转基因育种提供技术支持。方法]采用农杆菌侵染花柱头法,在不同时间侵染对4个海岛棉品种进行遗传转化,分析其对海岛棉的成铃率,田间卡那霉素抗性率及PCR阳性株率的影响。结果]农杆菌侵染花柱头法的平均成铃率为12.60%,田间卡那霉素检测的平均抗性率为9.87%,PCR鉴定的平均阳性株率为1.72%。花后12 h进行转化,成铃率、抗性率和阳性株率均最高,分别为20.25%、11.68%和2.59%。结论]在花后12 h利用农杆菌侵染花柱头法进行海岛棉遗传转化,转化效率最高。

Agrobacterium Infecting Flower Stigma for Sea-island Cotton(Gossypium barbademe L.) Genetic Transformation

Objective]The purpose of this project aimed to provide technological support for the Seaisland cotton transgenic breeding by exploring a more efficient and convenient genetic transformation technology.Method]Four Sea-island cotton varieties were used for genetic transformation with the method of agrobacterium infecting flower stigmas at different points of infection time.Then their cotton boll rate,field kanamycin resistance rate were counted,and the positive strains rate were analyzed by PCR.Result]The corresponding average rate were 12.60%,9.87%and 1.72%.Of four points of the infection time,12 hours after anthesis showed that the highest cotton boll rate,field kanamycin resistance rate and positive strain rate were 20.25%,11.68%and 2.59%respectively.Conclusion]The results showed that the best time of flower stigmata being infected with agrobacteria for Sea - island cotton genetic transformation was 12 hours after anthesis.