《植物生理学报》 2014, 50(4): 407-414

青杄PwUSP1基因的克隆及表达模式分析

崔晓燕¹, 李长江², 孙帆², 张世宏¹, 张凌云^2,*

¹吉林大学植物科学学院, 长春130062; ²北京林业大学森林培育与保护教育部重点实验室, 北京100083

通信作者：张凌云；E-mail: lyzhang73@sohu.com

摘　要：

广泛逆境胁迫蛋白(universal stress protein, USP)在非生物胁迫响应中起重要作用, 但在植物中其功能还大部分未知。本研究通过BLAST分析青杄EST文库, 得到USP基因的EST序列, 通过RACE PCR方法获取USP基因的末端序列, 经过与EST序列拼接得到USP基因的cDNA全长序列, 命名为PwUSP1。分析发现PwUSP1全长cDNA为1 167 bp, 编码区为519bp, 编码172个氨基酸残基。生物信息学分析显示, PwUSP1编码的蛋白相对分子质量为19.07 kDa, 理论等电点为6.38, 为非跨膜的亲水性蛋白。PwUSP1具有USP家族典型的UspA结构域和ATP结合位点G-(2X)-G-(9X)-G(S/T)。半定量RT-PCR 与RT-qPCR分析表明, PwUSP1在青杄的根、茎、针叶、花粉、种子中均有表达, 在根和花粉中表达量较高。同时, PwUSP1受干旱和盐胁迫的诱导表达上调, 均在处理6 h后表达量较高, 推测该基因可能在青杄逆境胁迫响应中发挥作用。

关键词：青杄; PwUSP1; 生物信息学分析; 组织表达; 胁迫响应

收稿：2013-12-09   修定：2014-01-27

资助：国家自然科学基金面上项目(31270663)和国家转基因生物新品种培育科技重大专项(2013ZX08009-003-002)。

Cloning and Expression Analysis of PwUSP1 from Picea wilsonii

CUI Xiao-Yan¹, LI Chang-Jiang², SUN Fan², ZHANG Shi-Hong¹, ZHANG Ling-Yun^2,*

¹College of Plant Sciences, Jilin University, Changchun 130062, China; ²Key Laboratory of Forest Silviculture and Conservation of the Ministry of Education, Beijing Forestry University, Beijing 100083, China

Corresponding author: ZHANG Ling-Yun; E-mail: lyzhang73@sohu.com

Abstract:

Universal stress proteins (USPs) play active roles in the abiotic stress responses of living bodies, but their functions remain largely unknown in plants. The full length cDNA of PwUSP1 was obtained by RACE PCR that spliced the terminal sequences of PwUSP1 with the EST based on the cDNA library of Picea wilsonii. The full length cDNA of PwUSP1 was 1 167 bp and the ORF was 519 bp which encoded 172 aa. Bioinformatics analysis showed that the theoretical molecular weight of PwUSP1 was 19.07 kDa and the isoelectric point was 6.38. PwUSP1 was a stable hydrophilic protein with non-transmembrane domain structure. PwUSP1 had the typical UspA domain of USP family with an ATP-binding site G-(2X)-G-(9X)-G(S/T). Semiquantitative RT-PCR and RT-qPCR analysis discovered that PwUSP1 was expressed in the root, stem, needle, pollen and seed of Picea wilsonii and was highly expressed in the root and pollen. PwUSP1 was up-regulated by drought and salt stresses and both was expressed most at 6 h after stress treatments, indicating that the PwUSP1 gene might play an important role in responses to some stresses.

Key words: Picea wilsonii; universal stress protein; bioinformatic analysis; tissue expression; stress response