《植物生理学报》 2010, 46(4): 354-358

茶树冷诱导基因RAV 的克隆与表达特性分析

陈林波^1,2, 李叶云¹, 王琴¹, 高永亮¹, 江昌俊^1,*

1安徽农业大学茶叶生物化学与生物技术教育部重点实验室, 合肥230064; 2云南省农业科学院茶叶研究所, 云南勐海 666201

通信作者：江昌俊；E-mail: jiangcj@ahau.edu.cn；Tel: 0551-5786982

摘　要：

对利用 cDNA-AFLP 技术所获得的茶树低温诱导差异表达片段 TDF, 通过 RACE 方法获得含完整编码区序列的茶树 RAV 基因 cDNA 克隆, 其开放阅读框编码 361 个氨基酸, 包含两个保守的结构域 AP2 和 B3, 与多种植物 RAV 蛋白具有高度 同源性。qRT-PCR 分析表明, 茶树 RAV 基因受低温、乙烯、NaCl 等上调表达, 最大表达量分别是诱导前的 5.8、10.0 和1.9 倍。在成熟叶片、芽、嫩茎中 RAV 基因表达量相近, 花蕾和嫩根中表达较低, 而在种子中不表达。推测该基因在组织 中的表达受到严格控制以及在响应非生物胁迫中发挥重要作用。

关键词：茶树; RAV转录基因; 基因克隆; qRT-PCR; 表达分析

收稿：2010-01-07   修定：2010-02-01

资助：国家科技支撑计划(2008BADC0B03)、国家自然科学基金(30871568)和安徽省自然科学基金(090411014)。

Cloning and Expression Analysis of RAV Gene Related to Cold Stress from Tea Plant [Camellia sinensis (L.) O. Kuntz]

CHEN Lin-Bo^1,2, LI Ye-Yun¹, WANG Qin¹, GAO Yong-Liang¹, JIANG Chang-Jun^1,*

¹Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education, Anhui Agricultural University, Hefei 230064,China; ²Tea Research Institute, Yunnan Academy of Agricultural Science, Menghai, Yunnan 666201, China

Corresponding author: JIANG Chang-Jun; E-mail: jiangcj@ahau.edu.cn; Tel: 0551-5786982

Abstract:

For a TDF, which has been gained from genes expressed differentially in tea plant under cold stress using cDNA-AFLP, containing a complete coding sequence cDNA was cloned by RACE, named CsRAV, and contains an ORF, which encodes a polypeptide of 361 amino acids including two conserved domains: AP2 and B3. Sequence alignment showed that CsRAV protein shared high identity with other plants. qRT-PCR results indicated that CsRAV was induced by cold, ethylene and NaCl, and maximum relative expression was 5.8, 10.0 and 1.9 times higher than before treatment, respectively. CsRAV gene expression level was closed in leave, bud and stem, and lower in flower and root, not detected in seed. It shows the expression of the CsRAV might be strictly controlled in different organs and plays an important role during abiotic stress in tea plant.

Key words: Camellia sinensis; RAV transcription factor; gene clone; qRT-PCR; expression analysis