农杆菌介导小麦遗传转化条件的优化

本研究以普通小麦品种扬麦15和Alondra's的幼胚产生的愈伤组织为受体材料,通过检测gus基因的瞬时表达情况,研究了农杆菌介导的主要影响因子的转化效率。研究结果表明,在相同条件下两个品种农杆菌侵染的最佳条件不尽相同。扬麦15的最佳优化条件为:先在不添加AS的培养基中预培养,然后在菌液浓度为OD600=0.2、AS浓度为100μmol/L条件下侵染10min,最后在25℃共培养1d;Alondra's则在菌液浓度为OD600=0.5、共培养时间为2d时,表现出最佳的gus基因瞬时表达率。为小麦的遗传转化提供参考。

Optimization of an Agrobacterium-mediated Transformation System for Wheat

In this research,we employed the callus induced from immature embryo of wheat varieties Yangmai 15 and Alondra's to be as materials for studying the transformation efficiency on main factors of A grobacterium-mediated for by using the transient expression frequency of gus gene.The results showed that the optimized condition for the above two varieties were different under the same condition.The best condition in Agrobacrerium-mediated transformation of Yangmai 15 are:First pre-cultured without acetosyringone,then infected for 10 min with A grobacterium tumefacien,bacterial at a concentration of OD600=0.2 and acetosyringone at a concentration of 100 μmol/L,finally co-cultured for 1 day at the temperature of 25℃;and under the similar condition,the most transient expression frequency of Gus gene in Alondra's with the concentration of Agrobacterium tumefaciens bacterial to OD600=0.5 and the co-culture to 2 days.This work would provide reference for the genetic transformation in wheat.