| 利用农杆菌介导法将PVA-CP基因导入马铃薯品种东农303的研究 |
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| 引用本文： | 于凤丽,卢翠华,李文斌,石瑛,邬震坤,梁彦涛.利用农杆菌介导法将PVA-CP基因导入马铃薯品种东农303的研究[J].中国马铃薯,2005,19(2):77-80. |
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| 作者姓名： | 于凤丽 卢翠华 李文斌 石瑛 邬震坤 梁彦涛 |
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| 作者单位： | 东北农业大学,黑龙江,哈尔滨,150030 |
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| 摘 要： | 以马铃薯早熟品种东农303的微型薯为受体材料,对其再生和农杆菌介导的遗传转化体系进行了研究,成功地将PVA-CP基因导入马铃薯中。结果表明:在加入5mg·L-1ZT和1mg·L-1IAA的培养基上微型薯再生频率最高达95%;农杆菌侵染薯块时,用浓度为OD600=0.5的菌液,侵染5min,共培养2d转化频率最高;在诱导芽阶段加入75mg·L-1的卡那霉素,采用延迟筛选的方法。得到的抗性芽有22株在含100mg·L-1卡那霉素的培养基上生根。经PCR和PCR-Southern杂交检测,15株为阳性植株。初步证明了PVA-CP基因已导入并整合到马铃薯品种东农303的基因组中。
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| 关 键 词： | 农杆菌 PVA-CP基因 马铃薯 遗传转化 |
| 文章编号： | 1672-3635(2005)02-0077-04 |
| 修稿时间： | 2005年1月5日 |
Introduction of PVA-CP Gene into Potato Cultivar Dongnong 303 by Agrobacterium-mediated Transformation |
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| YU Feng-li,LU Cui-hua,LI Wen-bin,SHI Ying,WU Zhen-kun,LIANG Yan-tao.Introduction of PVA-CP Gene into Potato Cultivar Dongnong 303 by Agrobacterium-mediated Transformation[J].Chinese Potato,2005,19(2):77-80. |
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| Authors: | YU Feng-li LU Cui-hua LI Wen-bin SHI Ying WU Zhen-kun LIANG Yan-tao |
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| Abstract: | Various factors influencing Agrobacterium-mediatad genetic-transformation and regeneration were investigated. The PVA-CP gene was introduced into microtuber explants of potato cultivar Dongnong 303. The highest frequency of regeneration was obtained on the medium with 5 mg·L-1 ZT and 1 mg·L-1 IAA. The optimum genetic-transformation system was as following: the Agrobacterium concentration was OD600 = 0.5; infection time, 5 min; co-cultivated time, 2 days; and the concentration of kanamycin used in resistant buds selection, 75 mg·L-1. Twenty two resistant buds rooted on the medium containing 100 mg·L-1 kanamycin. Results of PCR and PCR-Southern hybridization indicated that 15 regenerated plants were positive. These evidences suggested that the PVA-CP gene had been introduced and integrated into the genome of Dongnong 303. |
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| Keywords: | Agrobacterium tumefaciens PVA-CP gene potato genetic-transformation |
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