湖北农业科学 ›› 2020, Vol. 59 ›› Issue (12): 175-178.doi: 10.14088/j.cnki.issn0439-8114.2020.12.039

• 生物工程 • 上一篇    下一篇

棉花角斑病菌V8高效电转化条件的探索

王清1, 孟凡奇1, 张勇跃1, 缪卫国2, 刘志坚1   

  1. 1.河南省漯河市农业科学院,河南 漯河 462300;
    2.热带农林生物灾害绿色防控教育部重点实验室,海口 570228
  • 收稿日期:2019-07-26 发布日期:2020-08-26
  • 通讯作者: 刘志坚(1966-),男,河南漯河人,研究员,主要从事甘薯品种选育及利用工作,(电话)0395-3781568(电子信箱) liuzj66@126.com。
  • 作者简介:王 清(1988-),女,河南项城人,实习研究员,硕士,主要从事植物保护等工作,(电话)15890213237(电子信箱)1173823088@qq.com
  • 基金资助:
    国家甘薯产业技术体系项目(CARS-10-C-13); 国家重大基础研究计划项目(2011CB111612); 国家农业产业技术体系建设项目(CARS-34-GW8)

Exploring the conditions for efficient electric transformation of Xanthmonas campestris pv. malvacearum (Xcm) V8

WANG Qing1, MENG Fan-qi1, ZHANG Yong-yue1, MIAO Wei-guo2, LIU Zhi-jian1   

  1. 1. Luohe Academy of Agricultural Sciences,Luohe 462300,Henan,China;
    2. Key Laboratory of Green Preventionand Control ofTropical Plant Diseases and Pests(Hainan University), Ministry of Education,Haikou 570228,China
  • Received:2019-07-26 Published:2020-08-26

摘要: 对去甲基化氮胞苷(5-AZA)的最佳浓度以及影响棉花角斑病菌[Xanthmonas campestris pv. malvacearum (Xcm)]V8电转化效率的诸多因素进行逐一研究,以期获得高效的电转化条件。结果表明,选取100 μmol/L的5-AZA驯化处于对数中期的V8制备感受态细胞;保证电压2.1 kV、50 μg/mL质粒7 μL、4 ℃预冷复苏培养基SOC、振荡复苏培养4~5 h时,V8的电转化效率高达5×102/μg DNA。

关键词: 棉花角斑病菌[Xanthmona scampestris pv. malvacearum (Xcm)]V8, 菌株驯化, 电转化条件, 电转化效率

Abstract: In order to obtain efficient electrical conversion conditions, experimentswere conducted to study the efficiency between optimal concentration for 5-AZA and factors playing roles in the efficiency of cotton keratinae [Xanthmonas campestris pv. malvacearum (Xcm)]V8 one by one. The results showed that V8 for preparing competent cells was domestricated by 5-AZA with a concentration of 100 μmol/L at the logarithmic metaphase, Whiah was used to acclimate V8 in the medium logarithm to prepare competent cells. When the guaranteed voltage was 2.1 kV, 7 μL plasmid as well as concentration of 50 μg/mL, recovery media SOC of 4 ℃ pre-cooling, culturing with time of four to five hours by oscillating, the efficiency could reach about 5 ×102 /μg DNA.

Key words: Xanthmonas campestris pv. malvacearum (Xcm)]V8, domesticating strains, condition of electrical conversion, the efficiency of electrical conversion

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