陆地棉开花相关基因GhFLP1的克隆与功能验证  

Cloning and Functional Analysis of the Flowering-related Gene GhFLP1 from Upland Cotton(Gossypium hirsutum L.)

作　　者：张盼[1,2] 范术丽[2] 宋美珍[2] 庞朝友[2] 魏恒玲[2] 喻树迅[1,2] 

机构地区：[1]西北农林科技大学农学院,陕西杨陵712100 [2]中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南安阳455000

出　　处：《棉花学报》2016年第3期199-207,共9页Cotton Science

基　　金：国家科技支撑计划(2014BAD03B01)

摘　　要：为了明确陆地棉开花促进因子基因的作用,以中棉所36叶片基因组DNA和cDNA为材料克隆得到了开花相关基因Gh FLP1,分析了其特征及功能。该基因全长为537 bp,包含339 bp开放阅读框,编码112个氨基酸。预测分子量约为12.176 kDa,理论等电点为9.13。组织特异性表达分析表明,该基因在花蕾中优势表达,且在早熟品种(中棉所36、中棉所74)中表达量高于中熟品种(中棉所60、鲁棉研28)。启动子序列分析和外源激素处理实验表明该基因受水杨酸和赤霉素调控。农杆菌介导转化拟南芥发现,该基因能促使拟南芥莲座叶减少,开花期提前。荧光定量结果表明,在转基因拟南芥中,内源开花相关基因AtFT、AtLFY、AtAP1和AtSOC1表达量不同程度上调,AtFUL表达量基本不变,AtFLC表达量下调。研究结果显示该基因可能参与陆地棉开花时间的调控,为创制转基因早熟棉花新材料打下基础。To identify the function of flowering promoting factor genes, we used GhFLP1, a flowering related gene cloned from an upland cotton variety CCRI 36（Gossypium hirsutum L.）, for sequencing and function analysis. The full-length of the c DNA was 537 bp, and it includes a 339-bp open reading frame encoding a protein of 112 amino acids. Molecular mass was predicted to be 12.176 k Da, and the basic isoelectric point 9.13. Expression analysis in different tissues revealed the gene was strongly expressed in the flower bud, and higher expression levels were detected in premature varieties. A phytohormone treatment experiment revealed salicylic acid and Gibberellin positively regulated GhFLP1 in a 24-h time-course. Transgenic Arabidopsis plants carrying GhFLP1 exhibited significantly fewer rosette leaves and flowered earlier compared with the wild-type plants.q RT-PCRs suggested expression of AtFT, AtLFY, and AtAP1 was elevated, AtSOC1 and AtFUL expression was unchanged,and AtFLC expression was suppressed in transgenic Arabidopsis compared with the wild-type. The results preliminarily suggest an involvement of GhFLP1 in the regulation of flowering time, and provide further genetic resources for the breeding of short season cotton.

关 键 词：GhFLP1 GOSSYPIUM hirsutum 开花 基因克隆 基因表达 荧光定量检测 

分 类 号：S562[农业科学—作物学]