摘要
[Objective]The aimed to optimize transformation system for Agrobacterium-mediated green cotton shoot tip and provide reference for genetic engineering-assisted breeding of color cotton. [Method]With shoot tip of natural green cotton as receptor and by using Agrobacterium-mediated transformation method,the optimum conditions of regeneration system were obtained. [Result]Combining with vacuum infiltration,3-old-day shoot tips were infected for 10 minutes in Abarobacterium suspension with OD600 value about 0.5 followed by co-culture for 24 h on the medium containing MS＋l mg/L KT,pH value of 5.8. The regeneration culture was conducted in MS medium containing l mg/L KT and increasingly 30,50 and 70 mg/L Kan. The highest transformation frequency was 34.6%,and the positive resistant plantlets of green cotton 9804 were obtained. [Conclusion]Green cotton is more beneficial than white cotton to conduct genetic transformation.
[目的]对农杆菌法介导的绿色棉花茎尖转化体系进行筛选优化,为基因工程辅助彩棉育种提供参考。[方法]以天然绿色棉花的茎尖作为受体,采用农杆菌介导的转化方法,筛选绿棉茎尖再生体系的最佳条件,包括茎尖外植体最佳苗龄的筛选,激动素(KT)最适浓度的筛选,卡那霉素(Kan)敏感浓度的筛选,农杆菌菌液最佳侵染浓度的筛选以及真空渗透条件的优化筛选。[结果]3日龄的绿棉茎尖为最佳转化外植体;在OD600值为0.5的农杆菌菌液中侵染10min,同时辅助真空渗透的侵染效果最佳;在茎尖最适培养基MS+lmg/LKT,pH值5.8上共培养24h,转至Kan浓度梯度为30、50、70mg/L的筛选培养基上选择培养,茎尖分化苗阳性率达34.6%,并获得了绿棉9804的Kan阳性苗。[结论]绿色棉花比白色棉花更有利于进行遗传转化。
