张丽君 刘龙龙 张建珍 孙毅 康国帅 周建平 崔林

摘要

[目的] 優化农杆菌介导创伤胚转化的方法，提高转化率，培育具有特定功能的转基因品种，改善作物的生存能力和生态环境适应力。[方法]参照农杆菌划胚介导植物萌发种子基因转化方法，以燕麦、玉米和高粱的萌发种子为试验材料，农杆菌菌株LBA4404（含质粒P3301ubiAc）侵染转化燕麦、玉米和高粱的萌发种子，优化转化条件，探索燕麦、玉米和高粱遗传转化体系，探讨菌液浓度、种子萌发时间、超声波功率、乙酰丁香酮和草丁膦浓度对3种作物的影响并进行比较分析。[结果]农杆菌菌液浓度为0.6>OD600≥0.4 时转化效果最佳;第1次超声波功率为900 W，处理时间为10 min;第2次超声波功率为200 W，处理时间为4 min效果最佳。转化效率最高;添加乙酰丁香酮对燕麦、高粱和玉米的发芽有明显促进作用，但转化效果有明显区别;燕麦、玉米和高粱的转化植株对草丁膦的耐受性差异不大，与非转化植株相比，0.8 mg/L 除草剂胁迫下燕麦、玉米和高粱种子萌发受到一定程度的抑制。用1 mg/L 除草剂溶液对T₀和T₁代转化植株幼苗进行筛选，转化植株的存活率高于对照。[结论]经除草剂筛选和PCR对T₀和T₁代幼苗检测，初步证明获得了转化植株。

关键词 植物转化;燕麦;玉米;高粱

中图分类号 S188  文献标识码 A  文章编号 0517-6611（2015）03-025-04

Trauma Embryo of Genetic Transformation System with Agrobacterium Mediated Method

ZHANG Lijun1，2， LIU Longlong¹，ZHANG Jianzhen²， CUI Lin1* et al

（1. Crop germplasm resources research institute， Shanxi Academy of Agricultural Sciences; Key Laboratory of Crop Gene Resources and Germplasm Enhancement on Loess Plateau， Ministry of Agriculture， P. R. China， Taiyuan， Shanxi  030031;2. Research Institute of Applied Biology，Shanxi University，Taiyuan，Shanxi  030001）

Key words [Objective]The purpose was to optimize agrobacteriummediated plant germination seed transformation method，increase conversion rate， breed GM varieties with specific functions， and improve viability and adaptability of crops. [Method]This paper consulted agrobacteriummediated plant germination seed transformation method and selected germinating seeds of oat， maize and sorghum as test materials. Germinating seeds of oat， maize and sorghum were infected and transformed by agrobacterium strain， which included plasmid P3301ubiAc. Transformation conditions were optimized. Genetic transformation systems of oat， maize and sorghum were studied. Concentration of bacterium， seed germinating time， ultrasonic power， and influence of concentration of acetosyringone and glufosinate on the above three crops were compared and analyzed. [Result]The experiment showed that when 0.6>OD600≥0.4， transformation achieved the best effect; Ultrasonic power was 900 W at the first time and treatment time was 10 minutes; Ultrasonic power was 200 W at the second time and transformation achieved the best effect when treatment time was 4 minutes; Acetosyringone could affect germination of oat， sorghum and maize significantly. It could increase transformation rate of maize and lower that of sorghum， but for oat， its effect was not obvious; in terms of tolerance to glufosinate， the difference was not significant among transformed plants of oat， maize and sorghum. Compared with nontransformed plants， seed germination of oat， maize and sorghum were restrained to some degree under the stress of 0.8 mg/L herbicide solution. With 1 mg/L herbicide solution to select seedlings of T₀ and T₁ transformed plants， the result showed survival rates of transformed plants is higher than those of control group. [Conclusion ]After herbicide selection and PCR test on T₀ and T₁ seedlings， it was proved preliminarily that transformed plants were got.

Key words Plant transformation;Oat;Maize;Sorghum

基金项目 国家燕麦荞麦产业技术体系（CARS08A1）;山西省农科院博士基金项目（YBSJJ209）。

作者简介

张丽君（1981-），女，河北行唐人，助理研究员，博士，从事燕麦遗传育种研究。*通讯作者 。

收稿日期 20141203

随着生物技术的快速发展，植物基因工程在农作物品种改良中的应用越来越受到人们的重视，利用植物基因工程改良植物品质、增强抗性等是一种行之有效的方法，以弥补常规育种技术的不足，缩短育种周期。因此……