陆地棉开花相关基因GhFLP1的克隆与功能验证

发布时间：2018-11-04 17:11

【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。
[Abstract]:In order to clarify the role of flowering promoting factor gene in Upland cotton, the flowering related gene Gh FLP1, was cloned from the genomic DNA and cDNA of Zhongmianyi 36 leaves to analyze its characteristics and functions. The full length of the gene is 537 bp, which contains 339 bp open reading frame and encodes 112 amino acids. The predicted molecular weight is about 12.176 kDa, and the theoretical isoelectric point is 9.13. Tissue specific expression analysis showed that the expression of the gene was higher in early maturing varieties (Zhongmiansuo 36, Zhongmianyan 74) than in mid-maturing varieties (Zhongmiansuo 60, Lumianyan 28). Promoter sequence analysis and exogenous hormone treatment showed that the gene was regulated by salicylic acid and gibberellin. Agrobacterium tumefaciens mediated transformation of Arabidopsis thaliana showed that the gene could promote the decrease of rosette leaves and advance the flowering period. The results of fluorescence quantitative analysis showed that in transgenic Arabidopsis thaliana, the expression of endogenous flowering related gene AtFT,AtLFY,AtAP1 and AtSOC1 was up-regulated, the expression of AtFUL remained unchanged, and the expression of AtFLC was down-regulated. The results showed that the gene might be involved in the regulation of flowering time of upland cotton and lay the foundation for the creation of new transgenic early maturing cotton materials.
【作者单位】：西北农林科技大学农学院;中国农业科学院棉花研究所/棉花生物学国家重点实验室;
【基金】：国家科技支撑计划(2014BAD03B01)
【分类号】：S562

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