| 大豆农杆菌介导转化系统的优化研究 |
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| 引用本文： | 周思军,李希臣,刘昭军,刘丽艳,杨庆凯.大豆农杆菌介导转化系统的优化研究[J].东北农业大学学报,2001,32(4):325-331. |
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| 作者姓名： | 周思军 李希臣 刘昭军 刘丽艳 杨庆凯 |
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| 作者单位： | 1. 黑龙江省农科院生物技术研究中心,
2. 东北农业大学大豆所, |
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| 摘 要： | 对大豆子叶节再生系统及其农杆菌介导转化的适宜筛选剂浓度进行了优化 ;同时以发芽 5天的大豆无菌苗的下胚轴切段为外植体 ,以抗性愈伤为指标 ,对影响农杆菌介导的大豆转化频率的因子进行了研究。结果表明 :不同大豆品种对选择剂的敏感性存在一定差异。 4种选择剂比较 ,PPT抑制效果最好 ,品种之间比较稳定 ;潮霉素次之 ;卡那霉素的有效浓度最高。 4种选择剂的适宜浓度分别为 :卡那霉素 10 0～ 2 0 0 mg· L-1;G4 185 0～ 75 mg· L-1;潮霉素 2 0～ 30 mg·L-1;PPT5 mg· L-1。农杆菌感染时间和共培养时间均对大豆的转化有明显的影响。感染时间以 8分钟左右为宜 ;共培养时间以 2～ 4天为宜。在农杆菌感染和共培养阶段的乙酰丁香酮浓度和 p H值对大豆转化频率有明显影响。p H值以 5 .4 0～ 5 .6 0为宜 ;乙酰丁香酮的适宜浓度为 10 0 μmol· L-1左右。在感染农杆菌之前 ,让外植体在高渗培养基上预培养一段时间能提高大豆下胚轴切段的转化频率。最适预培养天数为 1d。
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| 关 键 词： | 大豆 农杆菌介导 转化 系统优化 |
| 文章编号： | 1005-9369(2001)04-0325-07 |
| 修稿时间： | 2001年4月9日 |
Optimization of the Agrobacterium-mediated transformation systems of soybean |
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| ZHOU Si jun ,LI Xi chen ,LIU Zhao jun ,LIU Li yan and YANG Qing kai.Optimization of the Agrobacterium-mediated transformation systems of soybean[J].Journal of Northeast Agricultural University,2001,32(4):325-331. |
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| Authors: | ZHOU Si jun LI Xi chen LIU Zhao jun LIU Li yan and YANG Qing kai |
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| Institution: | ZHOU Si jun 1,LI Xi chen 1,LIU Zhao jun 1,LIU Li yan 1 and YANG Qing kai 2 |
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| Abstract: | The regeneration system of soybean cotyledonary node and the suitable concentrations of selective agents in soybean cotyledonary node transformation were optimized.At the same time the factors influencing Agrobacterium-mediated soybean transformation were studied with the sections of soybean hypocoty1 from 5-day-old seedlings as explants and the resistant callus as target.The results show that:1)There is certain difference between the sensitivity of soybean cultivars to selective agents.Among the 4 tested agents,the inhibiting effect of PPT is the best and stable between cultivars;and hygromycin is the second.The effective concentration of kanamycin is the highest.The suitable concentrations of the 4 agents are kanamycin 100～200 mgl -1 ,G418 50～75mgl -1 ,hygromycin 20～30 mgl -1 and PPT 5 mgl -1 .2)The duration of infection and co-culture with Agrobacterium influences soybean transformation evidently.The suitable infection duration is about 8 minutes;and the appropriate co-culture duration is 2～4 days.3)Both the concentration of acetosyringone and pH value at stages of infection and co-culture influence the transformation frequency of soybean.The suitable pH value is between 5 40 and 5 60.The optimum concentration of acetosyringone is 100 μM.4)Preculture of explants,on hypertonic medium before infection of Agrobacterium can raise the transformation frequency of the sections of soybean hypocotyl.The optimun duration of preculture is one day.$$$$ |
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| Keywords: | soybean cryIA transformation transgenic plant |
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