青杄生长素抑制蛋白基因PwARP-1的克隆及表达分析

生物技术通报 ›› 2014, Vol. 0 ›› Issue (4): 64-70.

青杄生长素抑制蛋白基因PwARP-1的克隆及表达分析

孙帆, 罗朝兵, 周燕妮, 张凌云

(北京林业大学林学院森林培育与保护教育部重点实验室，北京 100083)

收稿日期:2014-01-17 出版日期:2014-04-29 发布日期:2014-04-29
作者简介:孙帆，男，硕士硕士生，研究方向：植物抗逆基因克隆和功能；Email：sunfan0041@126.com
基金资助:
国家自然科学基金面上项目(31270663)，国家转基因生物新品种培育科技重大专项(2013ZX08009003-002-004)

The Cloning and Expression Analysis of PwARP-1 in Picea wilsonii

Sun Fan, Luo Chaobing, Zhou Yanni, Zhang Lingyu

(Key Laboratory of Forest Silviculture and Conservation of Ministry of Education，College of Forestry，Beijing Forestry University，Beijing 100083)

Received:2014-01-17 Published:2014-04-29 Online:2014-04-29

摘要/Abstract

摘要： 从青杄的cDNA文库中筛选出一个生长素抑制蛋白基因PwARP-1(Auxin-repressed protein 1)，并通过RACE-PCR技术获得PwARP-1 cDNA全长。生物信息学分析显示，PwARP-1基因编码155个氨基酸残基的蛋白，分子量为17.1 kD，理论等电点为10.07，富含无规则卷曲。以多年生青杄和青杄幼苗为研究材料，通过RT-qPCR技术检测PwARP-1的组织特异性表达、激素的响应模式及种子萌发过程中的表达量模式。结果表明，PwARP-1在青杄各个组织中均有表达，在花粉中表达量最高，在种子中的表达量最低。PwARP-1在种子萌发过程中表达量呈先上升，在第10天表达量达到最高后开始下降。进一步试验表明，PwARP-1能响应多种逆境胁迫和激素处理。在赤霉素(GA)和生长素(IAA)处理下PwARP-1的表达量被抑制。在干旱胁迫和油菜素内酯(BR)激素处理下PwARP-1的表达量逐渐升高，而在在盐胁迫、茉莉酸甲酯(MeJA)和脱落酸(ABA)等激素处理下PwARP-1的表达量先下降后上升。这些结果显示PwARP-1可能参与了植物种子萌发、多种逆境胁迫和激素响应过程。

关键词: 青杄, 生长素抑制蛋白, 表达分析, 植物激素

Abstract: An auxin repressed protein gene PwARP-1 is cloned from Picea wilsonii. The full length cDNA of PwARP-1 is obtained by RACE-PCR assays based on the cDNA library of Picea wilsonii and EST fragment of PwARP-1. Bioinformatic tools are applied for the prediction of PwARP-1. PwARP-1 encodes a protein with 155 amino acids. The molecular weight is 17.1 kD and theoretical pI is 10.07 and the protein is rich in random coil. The tissue-specific expressions and the response to phytohormones of PwARP-1 were investigated by RT-qPCR assays. The tissue-specific expression assays suggested that the expression level in pollen was the highest among all tissues and the expression level in seed was the lowest. During seed germination, the expression of PwARP-1 was increased, peaked at the 10th day and then decreased. Furthermore, the expression of PwARP-1 was significantly inhibited by Auxin and Gibberellin. However, the expression level of PwARP-1 is up-regulated by Brassinolide and drought stress treatment. The expression level of PwARP-1 declined and then up-regulated under salt stress, methyl jasmonate, and abscisic acid treatments. These results indicate that PwARP-1 has a potential function in seed germination and responding to stresses and phytohormone treatments.

Key words: Picea wilsonii, Auxin-repressed protein, Expression analysis, Phytohormone

孙帆, 罗朝兵, 周燕妮, 张凌云. 青杄生长素抑制蛋白基因PwARP-1的克隆及表达分析[J]. 生物技术通报, 2014, 0(4): 64-70.

Sun Fan, Luo Chaobing, Zhou Yanni, Zhang Lingyu. The Cloning and Expression Analysis of PwARP-1 in Picea wilsonii[J]. Biotechnology Bulletin, 2014, 0(4): 64-70.

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