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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。
Soybean genetic transformation has been one of the difficulties in the field of plant gene transfer. Agrobacterium-mediated cotyledonary node method is the most commonly used method for soybean genetic transformation. The interaction of target cells and Agrobacterium is the key step of this method. Status is an important factor affecting the viability of target cells and thus the explant’s sensitivity to Agrobacterium. In this study, three varieties of Zhonghuang 10, Jack and Shuili Station were used for transformation experiments. According to the color and morphological changes of cotyledon and hypocotyl, the explants were divided into six states, and the efficiency of GUS transient expression was taken as the index The sensitivity of Agrobacterium tumefaciens to different target explants was compared in order to identify the optimal explant preparation period. The results showed that with the increase of germination time, there was a positive correlation between explant status and germination time and true leaf length, and the transient expression efficiency was significantly different. The best transient expression status of explants of three recipient varieties was different And Zhonghuang10 were the explants of HG, HG + CG and CG, and the explants of YL state at the best stage of Shuizhan variety. The results of this study are the standardization of soybean genetic transformation system Provided a theoretical basis.