摘要
【目的】克隆茶树CsAGL8基因序列,并探究该基因与茶树休眠的关系。【方法】以峨眉问春等茶树品种的休眠芽为材料,采用RT-PCR扩增MADS-box转录因子家族基因CsAGL8,并进行BLAST比对等生物信息学分析;采用实时荧光定量PCR检测CsAGL8在茶树8个组织和越冬期间的相对表达量;遗传转化拟南芥,对转基因植株进行表型观测及开花相关基因分析。【结果】CsAGL8的CDS全长为720 bp,编码239个氨基酸,含有高度保守的MADS-box结构域和半保守的K-box结构域,与拟南芥的AtAGL8相似性最高。CsAGL8编码区表现出一定的多态性,启动子区域存在较多的光、激素和胁迫响应等顺式作用元件。表达分析表明:CsAGL8在茶树不同组织中均有表达,在休眠顶芽中的表达量最高;越冬期间,CsAGL8的表达量呈先升高后降低的趋势,在特早生品种峨眉问春芽中的相对表达量显著低于对照品种,且下调时间早于对照品种。过表达CsAGL8,拟南芥转基因植株会早花10~15 d。【结论】CsAGL8可能参与了茶树越冬休眠调控。
[Purpose]To clone the CsAGL8 gene sequence of tea plant and to explore the relation-ship between this gene and tea plant dormancy.[Methods]The MADS-box transcription factor family gene CsAGL8 was cloned from the dormant buds of the tea cultivar‘Emei Wenchun’by RT-PCR,and bioinformatics analysis such as BLAST comparison was performed.The relative expres-sion levels in eight tissue samples and during overwintering were determined by qPCR.Genetic trans-formation of Arabidopsis thaliana,to observe the phenotype of transgenic plants and analyze the flowering-related genes.[Results]The CDS of CsAGL8 was 720 bp,which encoded 239 amino acids,and contained a highly conserved MADS-box domain and a semi-conserved K-box domain.CsAGL8 showed the highest similarity to AtAGL8 from A.thaliana.The coding region of CsAGL8 showed polymorphisms,and many cis-acting elements such as light,hormone,and stress response were found in the promoter region.Expression analysis showed that CsAGL8 was expressed in differ-ent tissues of tea plants,and the highest expression was detected in the dormant terminal buds.Dur-ing the overwintering process,the expression of CsAGL8 increased first and then decreased.Com-pared with the regular tea cultivar,the expression levels of CsAGL8 were not only significantly lower,but also down-regulated earlier in the buds of the extra-early cultivar‘Emei Wenchun’.Overexpres-sion of CsAGL8 in A.thaliana promoted early flowering for 10-15 days.[Conclusion]CsAGL8 may be involved in the regulation of winter dormancy in tea plants.
作者
胡小莉
付一丹
崔懂
夏心杰
陈盛相
唐茜
谭礼强
HU Xiaoli;FU Yidan;CUI Dong;XIA Xinjie;CHEN Shengxiang;TANG Qian;TAN Liqiang(College of Horticulture,Sichuan Agricultural University,Chengdu 611130,China;Tea Refining and Innovation Key Laboratory of Sichuan Province,Chengdu 611130,China)
出处
《云南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2023年第6期1032-1040,共9页
Journal of Yunnan Agricultural University:Natural Science
基金
国家自然科学基金项目(31800589)
大学生创新性实验计划项目(202010626132)
四川省科技计划项目(2022JDRC0035)。
作者简介
并列第一作者:胡小莉(1997-),女,重庆人,在读硕士研究生,主要从事茶树栽培与遗传育种研究。对本文贡献等同,E-mail:745836672@qq.com;并列第一作者:付一丹(2000-),女,河南濮阳人,在读硕士研究生,主要从事茶树栽培与遗传育种研究。对本文贡献等同,E-mail:2540113464@qq.com;通信作者:谭礼强(1987-),男,重庆人,博士,副教授,主要从事茶树遗传育种与栽培研究。E-mail:tanliqiang@sicau.edu.cn。
