氯化锂作为转基因大豆筛选剂的研究  

Lithium Chloride,an Alternative Selectable Marker for Soybean Transformation

作　　者：黄文佳[1] 李永光[1] 王志坤[1] 李维娜[1] 王涛[1] 王朋朋[1] 李文滨[1] 

机构地区：[1]东北农业大学大豆研究所/东北农业大学大豆生物学教育部重点实验室/农业部大豆生物学与遗传育种北方区域重点实验室,黑龙江哈尔滨150030

出　　处：《大豆科学》2012年第4期552-556,567,共6页Soybean Science

基　　金：国家转基因重大专项(2011ZX08004-002);大豆产业技术体系资助项目(CARS-04-PS04)

摘　　要：以东农50大豆子叶节为材料,以大豆子叶节丛生芽黄化率和丛生芽高度为考察指标,确定氯化锂作为转基因大豆筛选剂的最佳浓度和筛选临界观察时间。在此基础上,对转基因大豆丛生芽进行GUS染色,并对转化植株进行PCR检测。通过对受体植株进行氯化锂叶片涂抹,确定适宜的表型筛选浓度。结果表明,延迟7 d进行氯化锂筛选,筛选浓度为60 mmol.L-1为宜,最佳筛选天数为10 d。抗氯化锂丛生芽的GUS染色阳性率为80%,确定叶片涂抹法的适宜筛选浓度为700 mmol.L-1,并且经氯化锂筛选得到的转基因植株后代可以正常生长发育。The proper concentration of lithium chloride and selection time were determined by the yellowing rate and the height of cluster buds of Dongnong50,treated with different concentration of lithium chloride.The data showed the suitable selective concentration of lithium choride and treated time were 60 mmol·L-1 and 10 day,respectively.Further analysis revealed that 80% of cluster buds resistant to lithium chloride was positive after GUS dying.The transgenic plants were detected by PCR analysis.The result suggested the optimized concentration of lithium chloride was 700 mmol·L-1and the lineages of the transgenic plants could normally grow and develop.Results suggested that the selected marker of lithium chloride could be applied in rapid selection for transgenic soybean plants.

关 键 词：大豆 转基因植株 氯化锂 GUS染色 PCR 

分 类 号：S565.1[农业科学—作物学]