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作物学报 ›› 2013, Vol. 39 ›› Issue (12): 2162-2170.doi: 10.3724/SP.J.1006.2013.02162

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蔗ABA信号转导关键酶SoSnRK2.1基因的克隆与表达分析

谭秦亮1,李长宁1,2,杨丽涛1,2,*,李杨瑞1,2,*   

  1. 1 广西大学农学院 / 亚热带农业生物资源保护与利用国家重点实验室, 广西南宁530005; 2中国农业科学院甘蔗研究中心 / 农业部广西甘蔗生物技术与遗传改良重点实验室 / 广西作物遗传改良生物技术重点实验室 / 广西甘蔗遗传改良重点实验室, 广西南宁530007
  • 收稿日期:2013-01-29 修回日期:2013-06-09 出版日期:2013-12-12 网络出版日期:2013-09-29
  • 通讯作者: 杨丽涛, E-mail: liyr@gxu.edu.cn; 李杨瑞, E-mail: liyr@gxaas.net
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)课题(2013AA102604),国家国际合作项目(2009DFA30820, 0S2013ZR0130),广西自然科学基金创新团队项目(2011GXNSFF018002),广西科学研究与技术开发计划项目(桂科产1123008-1,桂科攻1222009-1B)和广西农科院团队项目(桂农科2011YT01)资助。

Cloning and Expression Analysis of Abscisic Acid Signal Transduction Key Enzyme Gene SoSnRK2.1 from Sugarcane

TAN Qin-Liang1,LI Chang-Ning1,2,YANG Li-Tao1,2,*,LI Yang-Rui1,2,*   

  1. 1 State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Agricultural College Guangxi University, Nanning 530005, China; 2 Sugarcane Research Center, Chinese Academy of Agricultural Sugarcane / Sugarcane Research Institute Guangxi Academy of Agricultural Sciences / Kev Laboratory of Sugarcane Biotechnology and Genetic Improvement (Guangxi), Ministry of Agriculture / Guangxi Crop Genetic Improvement and Biotechnology Laboratory / Guangxi Key Laboratory of Sugarcane Genetic Improvement, Nanning 530007, China
  • Received:2013-01-29 Revised:2013-06-09 Published:2013-12-12 Published online:2013-09-29
  • Contact: 杨丽涛, E-mail: liyr@gxu.edu.cn; 李杨瑞, E-mail: liyr@gxaas.net

摘要:

蔗糖非酵解型蛋白激酶(SnRK)是植物体内ABA信号转导途径的关键调控酶, 在植物的抗逆境生长过程中发挥着重要的作用本研究通过RT-PCRRACE-PCR技术克隆出编码甘蔗SnRK2蛋白的基因SoSnRK2.1该基因cDNA序列全长为1385 bp, 包含一个1002 bp的开放阅读框(ORF)。根据氨基酸序列预测SoSnRK2.1基因编码333个氨基酸残基, 与其他高等植物中相关蛋白的氨基酸具有高度的相似性, 尤其是与禾本科的玉米和水稻。构建该基因的原核表达载体pET-SoSnRK2.1, IPTG诱导下可得到38 kD 左右的蛋白, 与理论值一致。实时荧光定量PCR分析表明, SoSnRK2.1基因在ABA、干旱(PEG)+ABA、干旱(PEG)NaCl、低温(4℃)H2O2外源胁迫下均呈诱导表达的趋势。推测该基因参与调控干旱、高盐和低温等胁迫过程, 在甘蔗抗逆境胁迫中起重要作用。

关键词: 甘蔗, SoSnRK2.1, 基因克隆, 原核表达

Abstract:

Sucrose non-fermenting 1-related protein kinase (SnRK) is the key enzyme of ABA signal transduction pathways, which plays an important role in plant development under adversity environments. Reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) were applied to clone SoSnRK2.1. The cDNA full length of SoSnRK2.1 gene is 1385 bp, containing a 1002 bp complete open reading frame. The amino acids sequence analysis indicated that SoSnRK2.1 gene encodes 333 amino acids, sharing high homology with other species, especially gramineous of Zea mays and Oryza sativa. We constructed the gene prokaryotic expression vector pET-SoSnRK2.1, and obtained the protein of 38 kD induced by IPTG, which was consistent with the theoretical value. The result of Real time qPCR analysis showed that SoSnRK2.1 gene expression was basically up-regulated under different stresses of ABA, PEG+ABA, PEG, NaCl, cold, and H2O2.It indicated that SoSnRK2.1 gene takes part in regulations of drought, highsalt, low temperature or other stress process, which may play an important role in adaptation of plants to adverse stresses.

Key words: Sugarcane, SoSnRK2, Gene cloning, Prokaryotic expression

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