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利用根癌农杆菌将野芝麻叶片转化为转基因植株。

Transgenic plant production from leaf discs of Moricandia arvensis using Agrobacterium tumefaciens.

机构信息

Laboratory of Plant Breeding, Faculty of Agriculture, Tohoku University, 981, Sendai, Japan.

出版信息

Plant Cell Rep. 1996 Aug;15(11):799-803. doi: 10.1007/BF00233143.

Abstract

A high frequency shoot regeneration (80%) was developed from callus of leaf discs and stem internodes of Moricandia arvensis. Leaf discs were shown to be a preferable starting material for transformation experiments. Agrobacterium tumefaciens strain GV3101/pMP90 used in this study contained a binary vector with genes for kanamycin resistance, hygromycin resistance and β-glucuronidase (GUS). Maximum transformation efficiency (10.3%) was achieved by using kanamycin at the rate of 200 mg/l as a selection agent. Presence of tobacco suspension culture during co-cultivation and a pre-selection period of seven days after co-cultivation was essential for successful transformation. Transgenic plants grew to maturity and exhibited flowering in a glasshouse. GUS activity was evident in all parts of leaf and the presence of GUS gene in plant gemone was confirmed by PCR analysis.

摘要

高频 shoot 再生(80%)是从 Moricandia arvensis 的叶盘和茎节间的愈伤组织中开发的。叶盘被证明是转化实验的首选起始材料。本研究中使用的农杆菌菌株 GV3101/pMP90 含有一个带有卡那霉素抗性、潮霉素抗性和β-葡萄糖醛酸酶(GUS)基因的二元载体。使用 200mg/L 的卡那霉素作为选择剂,可实现最高转化效率(10.3%)。在共培养过程中存在烟草悬浮培养物和共培养后七天的预选择期对于成功转化至关重要。转基因植物生长成熟并在温室中开花。GUS 活性在叶片的所有部位都很明显,并且通过 PCR 分析证实了 GUS 基因在植物基因组中的存在。

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