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番茄果实LeNCED2基因的克隆及其表达分析
崔萌萌, 孙亮, 王灿磊, 赵胜利, 刘姝彤, 许小茜, 王艳萍, 纪凯, 李平, 冷平
0
(中国农业大学 农学与生物技术学院,北京 100193)
摘要:
为了解ABA对果实发育和成熟的调控作用,对与ABA合成相关的LeNCED2基因进行了克隆与分析。采用RT-PCR和RACE技术从番茄品种'嘉宝’(Solanum lycopersicum cv.Jiabao)果实中克隆得到了一个LeNCED1基因的同源片段,定名为LeNCED2(基因库注册号:EU912387)。该基因3'端序列为1 635 bp,与LeNCED1基因的DNA序列同源性为72.7%,与马铃薯中的StNCED2基因(AY662343)碱基序列同源性高达95%。碱基序列推导的蛋白序列与LeNCED1基因的同源性为80%。RT-PCR分析显示LeNCED2基因在番茄植株的根、茎、叶、花和萼片上都有表达。Real-time RT-PCR 分析显示LeNCED2基因在幼果期表达量较高,随着果实成熟逐渐降低,与ABA含量不一致。而LeNCED1基因在转色期表达量最大且与ABA含量相对应。
关键词:  番茄  LeNCED2基因  ABA  Real-time RT-PCR
DOI:10.11841/j.issn.1007-4333.2010.06.004
投稿时间:2010-04-26
基金项目:北京市科委重大资助项目(D0706002000091);中央高校基本科研业务费专项资金资助项目(2009-02-06)第一作者:崔萌萌,硕士研究生,E-mail:cuimeng123123@163.com通讯作者:冷平,教授,主要从事果品采后生理与分子生物学研究,E-mail:leng.p@263.net
Cloning and tissue expression of a new 9-cis-epoxycarotenoiddioxygenase gene LeNCED2 from tomato fruit
CUI Meng-meng, SUN Liang, WANG Can-lei, ZHAO Sheng-li, LIU Shu-tong, XU Xiao-qian, WANG Yan-ping, WANG Yan-ping, JI Kai, LI Ping
(College of Agronomy and Biotechnology,China Agricultural University,Beijing 100193,China)
Abstract:
A LeNCED1 homologous gene fragment named LeNCED2 was cloned from fruits of tomato (Solanum lycopersicum cv.Jiabao) by RT-PCR and RACE technology.The gene including 3' end is 1 635 bp,and it is 72.7% identity with LeNCED1 (Z97215) and 95% with potato StNCED2 gene (AY662343).The deduced amino acid sequence is over 80% identity with LeNCED1.RT-PCR analysis showed that the LeNCED2 expressed in roots,stems,leaves,and fuirts.Real-time RT-PCR analysis showed that the expression level of LeNCED2 was high in young fruits but decreased with fruit ripening progress.The expression pattern of LeNCED2,which was not in step with the ABA level,was not the same as the expression pattern of LeNCED1 which had highest level in turning stage of tomato fruits.
Key words:  tomato  LeNCED2 gene  abscisic acid  Real-time RT-PCR