| 擎天凤梨花器官全长cDNA文库的构建及EST分析 |
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| 引用本文： | 刘建新,沈福泉,田丹青,张智,葛亚英,沈晓岚,俞信英,王炜勇.擎天凤梨花器官全长cDNA文库的构建及EST分析[J].分子植物育种,2009,7(6):1137-1143. |
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| 作者姓名： | 刘建新 沈福泉 田丹青 张智 葛亚英 沈晓岚 俞信英 王炜勇 |
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| 作者单位： | 浙江省农业科学院花卉研究开发中心,杭州,311202 |
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| 基金项目： | 浙江省科技厅面上项目，浙江省重大科技专项重点项目，浙江省农业科学院博士科研启动费项目共同资助 |
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| 摘 要： | 为了通过基因工程手段来培育花色丰富、花型奇特的擎大凤梨新品种,从擎天凤梨中筛选出花色、花型相关调控基因则是十分必要的.本研究以擎天凤梨属品种Ostara为材料,采用SMART技术成功构建了擎天凤梨花器官的质粒型全长cDNA文库,初始文库滴度为3×10~6,插入片段平均长度大于1 kb;随机挑取2004个阳性克隆进行5'EST测序,获得高质量序列1 758条,经拼接获得1 365条单基因簇(unigene),其中跨叠群(contig)175个和单条序列(singlet)1 195个;经ORF寻找共获得有效ORF 1 283条;经COG分析EST序列编码的蛋白质被分为22类;经Blast分析后,获得一批花色、花器官发育、花期调控以及其它育种价值基因(包括cDNA全长或片段).该研究结果为擎天凤梨的花色、花型转基因等育种奠定了基础.
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| 关 键 词： | 擎天凤梨 cDNA文库 |
Construction of Full-Length cDNA Library from Floral Organ and EST Analysis in Guzmania |
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| Liu Jianxin,Shen Fuquan,Tian Danqing,Zhang Zhi,Ge Yaying,Shen Xiaolan,Yu Xinying,Wang Weiyong.Construction of Full-Length cDNA Library from Floral Organ and EST Analysis in Guzmania[J].Molecular Plant Breeding,2009,7(6):1137-1143. |
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| Authors: | Liu Jianxin Shen Fuquan Tian Danqing Zhang Zhi Ge Yaying Shen Xiaolan Yu Xinying Wang Weiyong |
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| Institution: | Liu Jianxin Shen Fuquan Tian Danqing Zhang Zhi Ge Yaying Shen Xiaolan Yu Xinying Wang Weiyong Research & Development Centre of Flower,Zhejiang Academy of Agricultural Sciences,Hangzhou,311202 |
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| Abstract: | To breed new species with abundant flower color and peculiar flower form by gene engineering methods,it is necessary to obtain some controlling genes, which correlated in the color and form of flower in guzmania. In the research, one kind of guzmania species named ostara was employed to be as material, and the full-length cDNA plasmid library of floral organ was constructed successfully by using SMART technology. The library had 3×10~6 original titer and more than 1 kb insert fragments. After 5'EST sequencing from 2004 positive clone chosen at random and clustering analyse, 1 758 high-quality sequences and 1 365 unigenes including 175 contigs and 1 195 singlets were obtained. These unigenes had 1 283 valid ORFs (open reading frames). COG analysis showed that those proteins coded by EST sequences were divided into 22 classes. Through blast analysis, full length or part cDNA of some genes controlling flower color, development of floral organs, florescence regulation and other breeding value genes were obtained. These results provided a fundament for guzmania flower color and form transgenic breeding. |
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| Keywords: | SMART EST COG Guzmania (Guzmania Ruiz & Pav) cDNA library SMART EST COG |
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