《植物生理学报》 2015, 51(8): 1307-1314

青杄PwUSP2基因的克隆和表达分析

周燕妮, 李艳芳, 张通, 张凌云^*

北京林业大学林学院, 省部共建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；Tel: 010-62336044

摘　要：

广泛逆境胁迫蛋白(USPs)参与碳缺乏、缺氧、干旱和高盐等多种非生物胁迫, 但在植物中的研究尚不深入。本文通过RACE-PCR的方法获得青杄PwUSP2基因的cDNA全长, 共987 bp, 其中编码区723 bp, 共编码240个氨基酸。利用生物信息学工具对其理化性质、二级结构和三级结构进行分析, 结果显示, 该蛋白理论分子质量为26.84 kDa, 理论等电点为4.61, 有丝氨酸和苏氨酸结合位点, 为非跨膜的亲水蛋白。PwUSP2具有USP家族典型的UspA结构域, 但无典型的ATP结合位点G-2X-G-9X-G[S/T]。RT-qPCR分析表明, PwUSP2在青杄花粉、果实、种子、成熟叶、幼叶、成茎中均有表达, 在果实中表达量较高。同时, PwUSP2在脱落酸(ABA)、茉莉酸甲酯(MeJA)等非生物胁迫下表达量有明显变化, 推测PwUSP2可能参与青杄对逆境胁迫的响应。

关键词：青杄; PwUSP2; 组织表达; 胁迫响应

收稿：2015-03-03   修定：2015-08-03

资助：国家自然科学基金项目(31270663)和国家转基因生物新品种培育科技重大专项(2014ZX08009-003-002)。

Cloning and Expression Analysis of PwUSP2 from Picea wilsonii

ZHOU Yan-Ni, LI Yan-Fang, ZHANG Tong, ZHANG Ling-Yun^*

Key Laboratory of Forest Silviculture and Conservation of the Ministry of Education, College of Forestry, Beijing Forestry University, Beijing 100083, China

Corresponding author: ZHANG Ling-Yun; E-mail: lyzhang73@sohu.com; Tel: 010-62336044

Abstract:

Universal stress proteins (USPs) involve in multiple abiotic responses including carbon starvation, O₂ deprivation, drought and high salinity; however, it still remains elusive in plants. In this study, the full length cDNA of PwUSP2 was obtained using RACE-PCR method. Bioinformatics analysis showed that the full length cDNA of PwUSP2 is 987 bp, including the open reading frame (ORF) 723 bp. The PwUSP2 contains 240 amino acids with a theoretical molecular weight of 26.84 kDa and theoretical isoelectric points (pI) of 4.61. It is also a hydrophilic protein with serine and threonine binding sites and has non-transmembrane domain. Meanwhile, it has the typical UspA domain of USP family but not the characteristic of ATP-binding site G-2X-G-9XG[S/T]. RT-qPCR analysis showed that PwUSP2 was expressed in different tissues, while highly expressed in fruit. Furthermore, the expression of PwUSP2 significantly changed under abiotic stresses such as abscisic acid (ABA) and methyl jasmonate (MeJA), indicating that PwUSP2 might be involved in response to multiple stresses in plants.

Key words: Picea wilsonii; PwUSP2; tissue expression; stress response