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作 者:卢翠华[1] 李晶[1] 石瑛[1] 陈伊里[1] 王凤义[1] 吕文河[1] 秦昕[1] 苏俊峰[1]
出 处:《中国马铃薯》2003年第5期277-279,共3页Chinese Potato Journal
基 金:黑龙江省科技厅"十五"科技资助项目 ;编号 :GA0 1B10 1-0 4
摘 要:以马铃薯极早熟品种东农 30 3的试管薯为外植体 ,通过农杆菌介导法成功地将几丁质酶(PBch)基因导入马铃薯中。薯块培养基MS +1mg LNAA +2mg LZT ,共培养 3d后 ,转到含卡那霉素 5 0mg L ,头孢噻肟钠 2 0 0mg L的相同的培养基 ,待抗性芽长到 1~ 2cm时 ,转入含卡那霉素75mg L的生根培养基进行生根筛选。对获得的 4株转基因植株进行PCR检测及PCR Southern杂交检测 ,有 3株呈阳性 ,转化率为 3 2 %。Microtubers of NEA 303, an extra early variety, were used as explants,and chitinase gene was introduced into the variety by means of agrobacterium mediated transformation.Firstly,microtubers as receptors and plasmid pBch as donor were co cultured for 3 days in MS + NAA 1mg/+ZT 2mg/L, then were transferred to the same medium plus kanamycin 50 mg/L and cefotaxime 200 mg/L.When shoots were 1~2 cm in length, they were transferred to radication medium containing kanamycin 75 mg/L for rhizogenesis.Tests of PCR and PCR Southern hybridization were made on 4 transgenic plants and 3 of those were positive reaction.Transformation rate was 3 2% in this study.
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