| SNAC1基因作为筛选标记基因用于棉花的遗传转化 |
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| 引用本文： | 李雪林,刘冠泽,聂以春,郭小平,张献龙.SNAC1基因作为筛选标记基因用于棉花的遗传转化[J].棉花学报,2010,22(1):36-41. |
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| 作者姓名： | 李雪林 刘冠泽 聂以春 郭小平 张献龙 |
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| 作者单位： | 1. 华中农业大学作物遗传改良国家重点实验室,武汉,430070;河南科技大学农学院,洛阳,471003
2. 华中农业大学作物遗传改良国家重点实验室,武汉,430070 |
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| 基金项目： | 国家高技术研究发展计划项目(2006AA00105);;国家棉花产业创新体系 |
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| 摘 要： | 以SNAC1基因作为筛选标记基因,NaCl作为筛选剂,通过农杆菌介导法将SNAC1和GUS基因导入棉花细胞并得到胚性愈伤组织。经过PCR检测证实,外源基因已经整合到棉花基因组中,GUS染色证明GUS基因得到表达。研究了NaCl作为棉花转化细胞的筛选剂在农杆菌介导转化中的应用浓度及方法,即NaCl的筛选浓度在1.1%～1.5%(W/V)之间,在愈伤组织诱导初期适当低一点,随着愈伤组织的生长而加大筛选浓度。由于NaCl不利于胚的分化,经过2～3次继代筛选后要及时去除NaCl以促进胚的分化。
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| 关 键 词： | 棉花 转化 SNAC1基因 NaCl |
Application of SNAC1 Gene as a Selection Marker Gene for Cotton (Gossypium hirsutum L.)Transformation |
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| LI Xue-lin,LIU Guan-ze,NIE Yi-chun,GUO Xiao-ping,ZHANG Xian-long.Application of SNAC1 Gene as a Selection Marker Gene for Cotton (Gossypium hirsutum L.)Transformation[J].Cotton Science,2010,22(1):36-41. |
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| Authors: | LI Xue-lin LIU Guan-ze NIE Yi-chun GUO Xiao-ping ZHANG Xian-long |
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| Institution: | 1.National Key Lab of Crop Genetic and Improvement/a>;Huazhong Agricultural University/a>;Wuhan 430070/a>;China/a>;2.College of Agriculture/a>;Henan University of Science and Technology/a>;Luoyang/a>;Henan 471003/a>;China |
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| Abstract: | Employing SNAC1 gene as a selection marker gene and NaC1 as a selection agent, SNAC1 and GUS genes were intro-duced into cotton genome via Agrobacterium. Transgenic calli were confirmed by PCR analysis, and expression of the GUS gene was showed with GUS staining. The concentration and method for employing NaC1 as a selection agent had been studied. The reasonable selection concentration of NaC1 should be 1.1%～1.5%(W/V). The start concentration of NaC1 in callus induction medium should be lower and the concentration increased as calli proliferated. Because NaC1 is not beneficial for embryo differ-entiation and development, NaC1 should be removed from the culture medium after the calli were subcultured for 2～3 times. |
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| Keywords: | cotton transformation SNAC1 gene NaCl |
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