北京林业大学学报2024,Vol.46Issue(10) :74-80.DOI:10.12171/j.1000-1522.20240055

'京枣39'愈伤组织遗传转化体系构建

Construction of genetic transformation system of'Jingzao 39'callus

冯雪晶 马灵 杨爽 薄文浩 陈学勋 庞晓明
北京林业大学学报2024,Vol.46Issue(10) :74-80.DOI:10.12171/j.1000-1522.20240055

'京枣39'愈伤组织遗传转化体系构建

Construction of genetic transformation system of'Jingzao 39'callus

冯雪晶 1马灵 1杨爽 1薄文浩 1陈学勋 2庞晓明1
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作者信息

  • 1. 林木遗传育种全国重点实验室,林木育种与生态修复国家工程研究中心,北京林业大学生物科学与技术学院,北京 100083
  • 2. 内蒙古敖汉旗双井林场,内蒙古赤峰 024308
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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 4d,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 4d;(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 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.

关键词

'京枣/39'/愈伤组织/遗传转化/eYGFPuv

Key words

Ziziphus jujuba'Jingzao 39'/callus/genetic transformation/eYGFPuv

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基金项目

国家重点研发计划(2022YFD2200404)

中央引导地方科技发展专项()

出版年

2024
北京林业大学学报
北京林业大学

北京林业大学学报

CSTPCD北大核心
影响因子:1.237
ISSN:1000-1522
参考文献量14
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