结缕草组织培养及农杆菌介导转化的主要因子优化

Optimization of major factors for tissue culture and Agrobacterium-mediated genetic transformation of Japanese lawngrass (Zoysia japonica)

以破除休眠的结缕草种子为试材,对其愈伤组织的诱导、分化及农杆菌介导法转化的主要因子作了研究。结果表明,结缕草种子愈伤组织的诱导以含2,4 D1mg/L+NAA3mg/L、分化以含6 BA3mg/L+KT3.5mg/L、生根以含NAA0.3mg/L的MS培养基为佳。GUS瞬间表达研究表明,以继代培养2周的愈伤组织为起始材料,采用负压处理和农杆菌感染10min,尔后在光条件下共培养转化为佳。

The major factors for callus induction, callus differentiation, and Agrobacterium-mediated transformation were studied in the current paper, using dormancy-broken seeds as the experiment material. The results indicated that the optimal culture medium for callus induction is MS containing 2,4-D 1 ( mg/L) and NAA 3 ( mg/L), for callus differentiation is MS containing 6-BA 3 ( mg/L) and KT 3.5 ( mg/L), and for root formation is MS containing 0.3 ( mg/L) NAA. The GUS transient expression study indicated the optimised factors for genetic transformation were as follows:Use 2-week-recultured calluses as the original material, infect for 10 minutes in Agrobacterium solution under negative pressure treatment, then follow with mutual culture under continuous light.