【作者】 郝心愿； 曹红利； 王新超； 杨亚军； 马春雷；

【Author】 HAO Xin-yuan;CAO Hong-li;WANG Xin-chao;YANG Ya-jun;MA Chun-lei;Tea Research Institute of Chinese Academy of Agricultural Sciences,National Center for Tea Improvement,Key Laboratory of Tea Plant Biology and Resources Utilization,Ministry of Agriculture;College of Horticulture,Northwest A&F University;

【机构】 中国农业科学院茶叶研究所/国家茶树改良中心/农业部茶树生物学与资源利用重点实验室； 西北农林科技大学园艺学院；

【摘要】 生长素响应因子(ARFs)是调控生长素应答基因表达的转录因子,在生长素信号转导中起着重要作用。本试验以茶树龙井43为材料,采用RT-PCR、SMART-RACE-PCR技术从茶树休眠芽中获得了茶树生长素响应因子基因的全长cDNA序列(命名为CsARF1)。并利用实时定量PCR方法(qRT-PCR)检测了该基因在茶芽进入休眠到萌发后不同时期的表达情况。结果显示:该基因全长为3222bp,包含2463bp的开放阅读框(ORF),编码820个氨基酸残基。预测该基因编码蛋白分子量为49.35kD,具有保守的N端DNA结合域和C端二聚化结构域,中间区域富含谷氨酸、丝氨酸和亮氨酸,是一个定位于细胞质的具有激活转录功能的可溶性蛋白。相似性分析结果表明,该基因编码的氨基酸序列与葡萄的相似性最高(83%)。实时荧光定量PCR分析显示,该基因在茶树越冬芽深休眠和萌动期表达量较高,说明该基因与茶树芽的休眠及解除密切相关。更多还原

【Abstract】 Auxin response factors(ARFs) are transcription factors that regulate the expression of auxin response genes,playing important roles in signal transduction of auxin.In this experiment,tea plants of "longjing 43" were employed.The full-length cDNA of ARF named CsARF1 was cloned by RT-PCR and SMART-RACE-PCR,followed by comprehensive putative protein sequence analysis,and the expression analysis of CsARF1 was conducted by qRT-PCT during the different phases of bud dormancy and budbreak.The results indicated that the full-length cDNA of CsARF1 is 3222 bp containing 2463 bp ORF which encodes 820 amino acid residues with a putative molecular mass of 49.35 kD.The soluble protein encoded by CsARF1 functions in the cytoplasm and consists of an amino-terminal DNA-binding domain,a middle region that functions as an activation domain,and a carboxy-terminal dimerization domain.The BLAST results indicated that the protein encoded by CsARF1 shared the highest identity(83%) with ARF in Vitis vinifera.The results of qRT-PCT showed that CsARFl had a marked rise in the expression level at deep dormant stage and sprouting stage,which demonstrates CsARF1 is relevant in the regulation of tea plant bud dormancy and budbreak.更多还原