| 绿色荧光蛋白基因转化柑橘胚性愈伤组织与高效再生 |
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| 引用本文： | 刘歆,段艳欣,范净,邓秀新,郭文武.绿色荧光蛋白基因转化柑橘胚性愈伤组织与高效再生[J].果树学报,2006,23(6):801-804,F0003. |
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| 作者姓名： | 刘歆 段艳欣 范净 邓秀新 郭文武 |
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| 作者单位： | 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： | 国家自然科学基金
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教育部科学技术研究项目
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霍英东教育基金
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国际科学基金(IFS) |
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| 摘 要： | 采用根癌农杆菌介导法对13份柑橘胚性愈伤组织进行绿色荧光蛋白基因(gfp)的转化,短时间内获得了温州蜜柑、橘橙杂种GWZ等11份柑橘材料的82个转化细胞系,其中佛罗斯特脐橙和日辉橘最易转化,分别获得28个和25个转化细胞系。选取3份材料的不同转化细胞系进行gfp的PCR分析,均获得了预期的gfp片段。研究表明,GFP标记可以在转化早期快速检测并分离转化子,及时剔除逃逸体,获得纯合的转化系;高水平的gfp表达不影响转化细胞增殖、生长和分化。这些带有GFP标记的转化细胞系为柑橘的有关基础研究提供了良好的试材,目前正用于柑橘体细胞杂交和不同倍性愈伤组织的社会化控制现象与机理等研究。
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| 关 键 词： | 柑橘 胚性愈伤组织 根癌农杆菌介导法 绿色荧光蛋白(GFP) |
| 文章编号： | 1009-9980（2006）06-801-04 |
| 收稿时间： | 2006-04-11 |
| 修稿时间： | 2006-04-112006-09-04 |
Transfer of green fluorescent protein gene into citrus embryogenic calluses and high efficient regeneration |
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| LIU Xin,DUAN Yan-xin,FAN Jing,DENG Xiu-xin,GUO Wen-wu.Transfer of green fluorescent protein gene into citrus embryogenic calluses and high efficient regeneration[J].Journal of Fruit Science,2006,23(6):801-804,F0003. |
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| Authors: | LIU Xin DUAN Yan-xin FAN Jing DENG Xiu-xin GUO Wen-wu |
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| Institution: | National Key Laboratory of Crop Genetic Improvement, Hnazhong A gricultural University, Wuhan, Hubei 430070 China |
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| Abstract: | Transformation of green fluorescent protein gene (gfp) into citrus embryogenic calluses of 13 cultivars mediated by Agrobacterium tumefaciens was conducted. Totally 82 transgenic lines from 11 cultivars were produced, among which 28 and 25 transgenic lines were obtained from Frost navel orange and Sunburst tangerine respectively. Further PCR analysis of 3 selected transformed cultivars amplified the expected GFP fragment. This study proved that GFP as a vital marker could localize the transgene sites at an early stage, be helpful for discriminating escapes and chimeras, and physically separating in vitro putative transformants. High level of gfp expression did not affect transgenic cell division, somatic embryogenesis and further differentiation. These transgenic cell lines are valuable materials in citrus basic research and currently being used in studies on citrus somatic fusion and social control mechanism between calluses of different ploidy levels. |
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| Keywords: | Citrus Embryogenic callus Agrobacterium tumefaciens Green fluorescence protein (GFP) |
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