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    冯雪晶, 马灵, 杨爽, 薄文浩, 陈学勋, 庞晓明. ‘京枣39’愈伤组织遗传转化体系构建[J]. 北京林业大学学报. DOI: 10.12171/j.1000-1522.20240055
    引用本文: 冯雪晶, 马灵, 杨爽, 薄文浩, 陈学勋, 庞晓明. ‘京枣39’愈伤组织遗传转化体系构建[J]. 北京林业大学学报. DOI: 10.12171/j.1000-1522.20240055
    shu
    Feng Xuejing, Ma Ling, Yang Shuang, Bo Wenhao, Chen Xuexun, Pang Xiaoming. Construction of genetic transformation system of ‘Jingzao 39’ callus[J]. Journal of Beijing Forestry University. DOI: 10.12171/j.1000-1522.20240055
    Citation: Feng Xuejing, Ma Ling, Yang Shuang, Bo Wenhao, Chen Xuexun, Pang Xiaoming. Construction of genetic transformation system of ‘Jingzao 39’ callus[J]. Journal of Beijing Forestry University. DOI: 10.12171/j.1000-1522.20240055
    shu

    ‘京枣39’愈伤组织遗传转化体系构建

    Construction of genetic transformation system of ‘Jingzao 39’ callus

      摘要
    • 摘要:
      目的 以‘京枣39’愈伤组织为外植体进行遗传转化，建立枣树愈伤组织遗传转化体系，优化阳性植株鉴定方法。
      方法 本研究以‘京枣39’愈伤组织为转化材料，利用含有报告基因 eYGFPuv 的改造载体，并通过根癌农杆菌介导方法转化，探究预培养时间、农杆菌菌液浓度、侵染时间、乙酰丁香酮浓度、共培养时间对转化效率的影响，建立愈伤组织遗传转化体系并优化阳性植株鉴定方法。
      结果 （1）卡那霉素对愈伤组织的最小致死质量浓度为30 mg/L。（2）遗传转化最佳处理条件为预培养时间为4 d，菌液OD600为0.6，侵染时间20 min，乙酰丁香酮浓度100 μmol/L，共培养时间为4 d；（3）阳性愈伤组织在365 nm紫外光下呈现明亮的荧光绿色，平均转化率达21.2%。
      结论 经对比试验，成功建立了‘京枣39’愈伤组织遗传转化体系，为加速枣遗传转化提供了新的方法。

       

      Abstract:
      Objective This paper aims to establish a genetic transformation system of jujube callus and optimize the identification method for selecting positive plants using the callus of ‘Jingzao 39’ as explants.
      Method Using ‘Jingzao 39’ callus as plant material, the modified vector containing the reporter gene eYGFPuv was transformed by agrobacterium tumefaciens mediated method, genetic transformation was performed by agrobacterium tumefaciens mediated method, the effects of pre-culture time, agrobacterium concentration, infection time, acetosyringone (AS) mass concentration and co-culture time on the genetic transformation of ‘Jingzao 39’ were investigated to establish a genetic transformation system of jujube callus and optimize the identification method for selecting positive plants.
      Result (1) The minimum lethal concentration of kanamycin on callus was 30 mg/L. (2)The optimal treatment conditions of genetic transformation included that pre-culture time was 4 d, bacterial solution concentration OD600 was 0.6, infection time was 20 min, AS mass concentration was 100 μmol/L, co-culture time was 4 d. (3) The positive callus group showed bright fluorescent green under 365 nm ultraviolet light and the transformation rate was averaged 21.2%.
      Conclusion Through comparative experiments, the ‘Jingzao 39’ callus genetic transformation system is successfully established, which provides a new method for accelerating the genetic transformation of jujube.

       

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