荔枝GA信号途径基因LcPIF4的克隆及其功能分析Cloning and function analysis of GA signal pathway gene LcPIF4 in litchi
王弋;董晨;魏永赞;郑雪文;李伟才;
摘要(Abstract):
【目的】克隆荔枝GA信号途径的LcPIF4基因,并对其序列特征、表达特点、基因功能、亚细胞定位及互作蛋白进行研究。【方法】利用花穗RNA-seq数据对LcPIF4基因进行生物信息预测及分析,通过qRT-PCR对LcPIF4基因在不同组织的表达进行研究,并在表达水平上分析其对烯效唑的响应。通过拟南芥转基因株系的构建对其基因功能及亚细胞定位进行分析,同时利用酵母双杂分析其与Lc DELLA-1蛋白的互作。【结果】克隆的荔枝LcPIF4基因ORF长1 617 bp,编码539个氨基酸,且具有经典的APB结构域和HLH结构域。qRT-PCR结果表明,LcPIF4基因在花穗、叶片、果皮等器官表达水平较高,且烯效唑处理后其表达水平显著下降。转基因试验表明,在拟南芥中过表达LcPIF4基因可促进下胚轴生长,且过表达LcPIF4-YFP的转基因材料可在细胞核位置检测到荧光信号。酵母双杂交结果表明,LcPIF4与赤霉素途径阻遏蛋白LcDELLA-1存在直接的蛋白互作。【结论】荔枝LcPIF4蛋白具有拟南芥PIF4蛋白相同的结构域;LcPIF4在花穗中高表达,且其表达被烯效唑所抑制。LcPIF4可促进拟南芥下胚轴生长,其编码蛋白定位于细胞核。LcPIF4蛋白与Lc DELLA-1在酵母中存在互作。
关键词(KeyWords): 荔枝;LcPIF4;烯效唑;花穗;表达模式
基金项目(Foundation): 海南省自然科学基金(317241);; 广东省自然科学基金(2018A030307007);; 中央级公益性科研院所基本科研业务费专项(1630062018013);; 国家现代农业(荔枝龙眼)产业技术体系(CARS-032)
作者(Author): 王弋;董晨;魏永赞;郑雪文;李伟才;
Email:
DOI: 10.13925/j.cnki.gsxb.20180139
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