耐盐芦苇高频遗传转化体系的建立

Establishment of High Frequency Regeneration System of Phragmites australis

以耐盐芦苇(Phragmites australis)种子愈伤组织为材料,以根瘤农杆菌EHA105为媒介建立高效的芦苇遗传转化体系。结果表明:在潮霉素浓度为50 mg/L、6-BA浓度为0.8 mg/L时不定芽分化率达到最高。将沉默载体pFGC1008-GRPi通过根瘤农杆菌EHA105在芦苇愈伤组织中进行遗传转化,共获得8个抗性株系,转化率为3.9%。实时荧光定量PCR检测显示6个株系实现了基因沉默,表明该转化体系高效可用。

An efficient genetic transformation system of P.australis is established by using the callus of salt tolerant reed(P.australis)seeds as materials and Agrobacterium tumefaciens EHA105 as vector.The results show that the differentiation rate of adventitious buds reaches the highest with 50 mg/L Hyp and 0.8 mg/L 6-BA.The silencing vector pFGC1008-GRPi is transformed into reed callus by EHA105.A total of 8 positive transformation lines are ob-tained,with a transformation rate of 3.9%.Real time fluorescent quantitative PCR detection shows that 6 strains a-chieved gene silencing,indicating that the transformation system is convenient and available.