农杆菌介导的小麦幼胚遗传转化体系的优化

为了优化农杆菌介导的小麦幼胚转化体系,通过对4个小麦品种河农827、石新828、河农825、良星99的幼胚组织培养及再生系统的研究表明,在改良培养基MSW1和MSW2中,小麦幼胚胚性愈伤组织诱导率与MSW0培养基相比分别提高了6.0%和5.4%,并且显著提高了植株再生能力。采用混合正交试验设计,对农杆菌遗传转化过程中小麦幼胚的诱导培养基、诱导时间、农杆菌侵染浓度、侵染时间进行了优化。结果表明,在诱导培养基MSW2,诱导培养6d,OD660=1,侵染时间3h时,愈伤GUS瞬时表达率及抗性愈伤成活率最高,对河农827成活的抗性再生植株进行PCR检测,获得了候选转基因植株。

Optimization of Wheat Immature Embroys Genetic Transformation System Mediated by Agrobacterium tumefaciens

Tissue culture regeneration system of wheat varieties Henong 827,Shixin 828,Henong 825 and Liangxing 99 were studied.The results indicated that MSW1 and MSW2 medium modified from MSW0 medium,could increase the induction frequency of embryogenic callus by 6.0% and 5.4%,respectively.Moreover,the frequency of plant regeneration increased significantly.With the aid of an orthogonal design,we optimized the method for Agrobacterium-mediated transformation of immature embryos of wheat.The highest survival rate of resistant callus and transient expression frequency of GUS gene were obtained for immature embryos induced for 6 days in MSW2 medium,followed by immersing in inoculation suspension of OD660=1 in darkness at 23-25℃ for 3 h.A candidate of henong 827 transgenic plants were obtained by PCR analysis.