蕙兰Flowering locus T基因的克隆及其对开花的影响.pdfVIP

中国农业科学 2013,46(7):1419-1425 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2013.07.012 蕙兰Flowering locus T基因的克隆及其对开花的影响 孙崇波，向 林，李小白，秦德辉，李伯钧，郭方其，吴 超 （浙江省农业科学院园艺研究所，杭州310021 ） 摘要：【目的】探讨CfFT 基因在蕙兰成花中的作用。【方法】采用RT-PCR 结合RACE 技术从蕙兰(Cymbidium faberi)中克隆Flowering locus T （FT）同源基因CfFT。采用实时定量RT-PCR 对不同组织及不同花发育时期CfFT 进行表达分析。将该基因克隆到PBI121 载体上导入烟草中，并对不同转基因烟草株系中的CfFT、NFL、NtFUL 和NAP1 基因进行实时定量RT-PCR 分析。【结果】对蕙兰花芽分化不同时期的CfFT 的表达分析表明，CfFT 在花芽分化初期的 表达量最高，之后随着花芽的成熟表达量逐渐降低。将CfFT 导入烟草进行异源表达，转基因株系表现出明显的早花 表型。对开花时间不一的转基因株系中的CfFT 表达分析表明，其表达量与转基因烟草开花时间早晚成正比。进一步 对这些株系内源的NFL、NAP1 和NtFUL 表达分析表明，NFL、NAP1 和NtFUL 基因的表达量与CfFT 表达成正比，说明 NFL、NAP1 和NtFUL 的表达受FT 基因的上游调控。【结论】在烟草中异源表达蕙兰中的CfFT 基因能促进烟草提前开花。 关键词：蕙兰；FT 基因；成花诱导；烟草；实时荧光定量表达 Isolation of Flowering Locus T Ortholog and the Effects on Blooming of Cymbidium faberi SUN Chong-bo, XIANG Lin, LI Xiao-bai, QIN De-hui, LI Bo-jun, GUO Fang-qi, WU Chao (Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021) Abstract: 【Objective 】The objective of this study was to analyze the function of Flowering locus T(FT) on blooming in Cymbidium faberi. 【Method 】 The full-length cDNA sequence of CfFT was obtained by RT-PCR and RACE cloning technology. The expression patterns of CfFT in different tissues and flower buds of different developmental stages were analyzed by real -time quantitative RT-PCR. To investigate the function of CfFT, an expression vector was constructed for transformation into tobacco by using agrobacterium-mediated method. 【Result 】In flowers, CfFT was expressed more in young flower buds than in mature flowers and was predominantly expressed in young ovary. Ectopic expression of CfFT in transgenic tobacco plants showed novel