农杆菌介导大豆子叶节转化法外植体选择新方法  

New Method for the Selection of Explants in the Agrobacterium Mediated Cotyledon Nodes Transformation in Soybean( Glycine max L.)

作　　者：王爽[1,2] 郭兵福[2] 张丽娟[2] 郭勇[2] 金龙国[2] 杨慧[2] 邱丽娟[2] WANG Shuang GUO Bing-fu ZHANG Li-juan GUO Yong JIN Long-guo YANG Hui QIU Li-juan(College of Agriculture, Northeast Agricultural University, Harbin 150030, China The National Key Facility for Crop Gene Resources and Genetic Improvement （NFCRI）/MOA Key Lab of Soybean Biology （Beijing）, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China)

机构地区：[1]东北农业大学农学院,黑龙江哈尔滨150030 [2]中国农业科学院作物科学研究所/国家农作物基因资源与遗传改良重大科学工程,北京100081

出　　处：《大豆科学》2016年第5期723-729,共7页Soybean Science

基　　金：转基因新品种培育重大专项(2014ZX08004-001)

摘　　要：大豆遗传转化一直是植物转基因领域的难点之一,农杆菌介导的子叶节法是大豆遗传转化最常用的方法,靶细胞与农杆菌的互作是该方法的关键环节,而外植体的状态是影响靶细胞活力进而影响外植体对农杆菌敏感性的重要因素。本研究利用中黄10、Jack和水里站3个品种材料开展转化试验,根据子叶和下胚轴的颜色和形态变化将外植体划分为6种状态,并以GUS基因瞬时表达效率为指标,比较不同状态外植体靶细胞对农杆菌敏感性,旨在鉴定外植体的最佳制备时期。结果表明:随着萌发时间的增加,外植体状态变化与萌发时间和真叶长度呈正相关,其瞬时表达效率差异显著,且3个受体品种外植体最佳瞬时表达状态存在差异,Jack和中黄10品种的最佳时期为HG、HG+CG、CG这3种状态的外植体,水里站品种的最佳时期YL状态的外植体,研究结果为大豆遗传转化体系的标准化提供了理论依据。The genetic transformation of soybean is one of the difficulties in plant transgenic and Agrobacterium-mediated cotyledonary node transformation method is the most commonly used in soybean genetic transformation. Interaction between target cells and Agrobacterium is the key of this method and explant status is an important factor that influence sensitivity and regeneration ability. In this study, the explant was divided into 6 types according to the color and status of cotyledon and hypocotyl. Transient transformation was carried out with zhonghuang 10, Jack and Shuilizhan and the index of transient expression efficiency of GUS gene was used to explore the best time to prepare explants. The results showed that the explants status were positively correlated with germination time and leaf length and had effect on transient expression efficiency. The best status of different receptors was different. The best periods of Jack and Zhonghuang 10 were HG, HG ＋ CG and CG, the best periods of Shuilizhan was YL Those cnclusions could provide a reference for the standardization of soybean genetic transformation system.

关 键 词：外植体状态 瞬时表达 萌发时间 

分 类 号：S565.1[农业科学—作物学]