《植物生理学报》 2018, 54(8): 1341-1348

草莓蔗糖非发酵-1-相关蛋白激酶1 (SnRK1) α亚基编码基因的克隆及表达分析

罗静静, 张亚飞, 张淑辉, 彭福田^*, 肖元松^*

山东农业大学园艺科学与工程学院, 作物生物学国家重点实验室, 山东泰安271018

通信作者：彭福田；E-mail: pft@sdau.edu.cn, ysxiao@sdau.edu.cn

摘　要：

植物的蔗糖非发酵-1-相关蛋白激酶1 (SnRK1)与酵母蔗糖非发酵蛋白激酶1 (SNF1)以及哺乳动物AMP激活的蛋白激酶(AMPK)同源, 都是异源三聚复合体结构, 含有α催化亚基和β、γ两个调节亚基来维持蛋白结构的稳定和激酶活性。本试验以‘妙香7号’草莓(Fragaria × ananassa)为材料, 通过反转录PCR克隆得到一个SnRK1的α催化亚基编码基因, 命名为FaSnRK1α。序列分析显示该基因全长1 557 bp, 共编码518个氨基酸, 预测FaSn-RK1α蛋白分子质量为59.159 kDa, 理论等电点为8.54, 定位于细胞质和细胞核。生物信息学分析发现FaSn-RK1α的氨基酸序列与其他植物SnRK1α蛋白具有较高同源性, 含有KD (kinase domain)、UBA (ubiquitin associated domain)和β-SID (β-submit interaction domain)三个保守结构域。组织特异性分析表明FaSnRK1α在草莓根、茎、叶、花和果实中均有表达, 在果实发育进程中FaSnRK1α的表达水平呈上升趋势。荧光定量PCR分析表明, 果实中FaSnRK1α受脱落酸(ABA)诱导, 表明该基因可能与ABA诱导的果实发育和成熟有关。

关键词：草莓; SnRK1; 脱落酸; 基因克隆; 表达模式

收稿：2018-05-02   修定：2018-08-15

资助：国家自然科学基金(31672099)、国家现代农业产业技术体系建设专项资金(CARS: 30-2-02)、山东省教育厅高等学校科技计划项目(J16LF54)和山东省“双一流”建设奖补资金(SYL2017YSTD10)。

Cloning and expression analysis of sucrose non-fermenting-1-related protein kinase 1 (SnRK1) α-subunit gene in strawberry

LUO Jing-Jing, ZHANG Ya-Fei, ZHANG Shu-Hui, PENG Fu-Tian^*, XIAO Yuan-Song^*

College of Horticulture Science and Engineering, Shandong Agricultural University; State Key Laboratory of Crop Biology, Taian, Shandong 271018, China

Corresponding author: PENG Fu-Tian; E-mail: pft@sdau.edu.cn, ysxiao@sdau.edu.cn

Abstract:

SnRK1 (sucrose non-fermenting-1-related protein kinase 1) is the plant ortholog of the budding yeast SNF1 (sucrose-non-fermenting 1) and mammalian AMPK (AMP-activated protein kinase). All three enzymes typically function as heterotrimeric complexes that require a catalytic α-subunit and regulatory β- and γ-subunits for protein stability and kinase activity. In this article a SnRK1 α-subunit gene was cloned from the leaves of strawberry (Fragaria × ananassa cv. Miaoxiang 7) using reverse transcription PCR, and was designated as FaSnRK1α. Sequence analysis showed that the total length of FaSnRK1α is 1 557 bp, encoding 518 amino acids. The molecular mass of FaSnRK1α protein is 59.159 kDa and the isoelectric point is 8.54, and FaSnRK1α locates in cytoplasm and nuclear. Bioinformatic analysis showed that FaSnRK1α protein shares high sequence identity with the reported plant SnRK1α proteins, with three conserved domains, annotated as KD (kinase domain), UBA (ubiquitin associated domain) and β-SID (β-submit interaction domain). Tissue specificity analysis showed that FaSnRK1α was expressed in root, stem, leaf, flower and fruit and has a rising trend during fruit development. Quantitative real-time PCR analysis demonstrated that the expression of FaSnRK1α was highly induced by abscisic acid (ABA), suggesting FaSnRK1α might play an important role in strawberry fruit development response to ABA.

Key words: strawberry; SnRK1; abscisic acid; gene cloning; expression pattern