华北农学报 ›› 2005, Vol. 20 ›› Issue (2): 12-15. doi: 10.3321/j.issn:1000-7091.2005.02.004

所属专题: 油料作物

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油菜下胚轴农杆菌介导法转化影响因素探讨

王景雪1, 杜建中2, 荣二花2, 李晓灿2, 孙毅2   

  1. 1. 中山大学教育部基因工程重点实验室, 广东, 广州, 510275;
    2. 山西省农业生物技术研究中心, 山西, 太原, 030031
  • 收稿日期:2004-08-03 出版日期:2005-04-28
  • 作者简介:王景雪(1961-),女,山西新绛人,研究员,博士,主要从事植物生物技术及基因工程研究工作.
  • 基金资助:
    山西省自然科学基金项目(2001096);山西省农科院资助项目

Study on the Major Factors Affecting Transformation Efficiency by Agrobacterium-mediated in Brassica napus

WANG Jing-xue1, DU Jian-zhong2, RONG Er-hua2, LI Xiao-can2, SUN Yi2   

  1. 1. The Key Laboratory of Gene Engineering of Ministry of Education, Zhongshan University, Guangzhou 510275, China;
    2. The Agro-biotechnology Research Center of Shanxi Province, Taiyuan 030031, China
  • Received:2004-08-03 Published:2005-04-28

摘要: 利用甘蓝型油菜下胚轴为外植体, 对油菜农杆菌介导转化中的主要因素进行了研究, 并由此建立了油菜下胚轴农杆菌介导的遗传转化体系.研究发现, 在农杆菌介导的油菜下胚轴转化中, 较好的转化体系是:外植体在农杆菌侵染前进行3 d的预培养.农杆菌的浸染浓度为OD600=0.2~0.4共培养时的pH值为5.2.在转化后的分化培养基中附加30μmol/L AgNO3.

关键词: 油菜, 转基因, 农杆菌介导法

Abstract: The hypocotyls segments were transformated by Agrobacterium mediated transformation in Brassica napus. Some factors affect shoot regeneration of transformated cells were studied. Our study revealed that, for Agrobacterium -mediated transformation, the hypocotyls segments were preconditioned on preconditioning medium for 3 days prior to infect with A.tumefaciens and the addition of 30 μmol/L AgNO3 in the regeneration medium were prerequiste for obtained high transformation efficiency of explants. Explants were submerged in the A.tumefaciens suspension at the concentration of OD600=0.2-0.4 for 5 minutes and placed on the co-cultivation medium pH 5.2 could result in high transformation efficiency of explants.

Key words: Brassica napus L., Transformation, Agrobacterium-mediated transformation

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引用本文

王景雪, 杜建中, 荣二花, 李晓灿, 孙毅. 油菜下胚轴农杆菌介导法转化影响因素探讨[J]. 华北农学报, 2005, 20(2): 12-15. doi: 10.3321/j.issn:1000-7091.2005.02.004.

WANG Jing-xue, DU Jian-zhong, RONG Er-hua, LI Xiao-can, SUN Yi. Study on the Major Factors Affecting Transformation Efficiency by Agrobacterium-mediated in Brassica napus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2005, 20(2): 12-15. doi: 10.3321/j.issn:1000-7091.2005.02.004.

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