《植物生理学报》 2017, 53(4): 591-599

小桐子JcYkt6基因的克隆和表达分析

黄尧瑶¹, 邓明华³, 龚明⁴, 陈浩维¹, 施卫省², 文锦芬^2,*

昆明理工大学¹现代农业工程学院, ²建筑与城市规划学院, 昆明650500; ³云南农业大学园林园艺学院, 昆明650201; ⁴云南师范大学生命科学学院, 昆明650500

通信作者：文锦芬；E-mail: wenjf888@163.com

摘　要：

以小桐子(Jatropha curcas) cDNA为模版, 克隆了JcYkt6基因的CDS序列。序列分析表明该基因包含600 bp完全阅读框(ORF), 编码199个氨基酸。预测其编码蛋白质的分子量为22.53 kDa, 等电点为6.90。Blast搜索结果及进化分析结果表明, JcYkt6蛋白与香瓜和黄瓜的Ykt6蛋白序列一致性最高(91%), 亲缘关系最近。该基因编码的蛋白具有一个典型的VAMP基元。定量分析结果显示, JcYkt6基因在小桐子根、茎、叶、花、果皮和种子中都有表达, 并且在种子中表达量最高。在种子生长发育的前期其表达水平不断增加, 在种子生长发育的后期(50 d)达到最高值, 然后下降。表达量变化过程与种子中储藏蛋白质合成趋势基本一致, 推测其参与小桐子种子储藏蛋白质的合成和运输。ABA、PEG、NaCl、4°C和机械损伤处理下JcYkt6表达水平都不同程度上调, 推测JcYkt6具有多种功能, 可能参与小桐子非生物胁迫响应和激素信号传导。

关键词：小桐子; SNAREs; JcYkt6; 克隆; 生物信息学分析; 表达分析

收稿：2016-11-09   修定：2017-03-14

资助：国家自然科学基金(31460355、31260064和31460059)。

Cloning and expression analysis of JcYkt6 gene in Jatropha curcas

HUANG Yao-Yao¹, DENG Ming-Hua³, GONG Ming⁴, CHEN Hao-Wei¹, SHI Wei-Sheng², WEN Jin-Fen^2,*

¹Faculty of Modern Agricultural Engineering, ²Faculty of Architecture and City Planning, Kunming University of Science and Technology, Kunming 650500, China; ³College of Landscape and Horticulture, Yunan Agricultural University, Kunming 650201, China; ⁴School of Life Sciences, Yunnan Normal University, Kunming 650500, China

Corresponding author: WEN Jin-Fen; E-mail: wenjf888@163.com

Abstract:

A complete CDS sequence of JcYkt6 was cloned from Jatropha curcas. Sequence analysis showed that the complete ORF of JcYkt6 was 600 bp, encoding 199 amino acid residues. Putative molecular mass of JcYkt6 was 22.53 kDa and isoelectric point (PI) was 6.90. The results of Blast and phylogenetic analysis showed that JcYkt6 had highest identity (91%) and closest relationship with Ykt6 protein of Cucumis melo and C. sativus. The gene encodes a protein with a typical VAMP motif. The JcYkt6 expressed significantly in different organisms and had the highest transcript profile in seed. The expression level of JcYkt6 increased in early stage of seed growth, and reached the highest value in late stage (50 d), then declined. Expression level of JcYkt6 in seed had the same trends with seed storage protein accumulation, indicating that JcYkt6 probably involved in the synthesis and transport of seed storage protein. The treatments of ABA, PEG, NaCl, 4°C and merchanical damage upregulated JcYkt6 gene expression, indicating that JcYkt6 also involved in response abiotic stress and singnal transduction process in J. curcas.

Key words: Jatropha curcas; SANREs; JcYkt6; clone; bioinformatics analysis; expression analysis