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Acta Agron Sin ›› 2015, Vol. 41 ›› Issue (03): 499-506.

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Cloning and Expression Analysis of CIPK Gene in Sugarcane

HUANG Long,SU Wei-Hua,ZHANG Yu-Ye,HUANG Ning,LING Hui,XIAO Xin-Huan,QUE You-Xiong*,CHEN Ru-Kai*   

  1. Key Laboratory of Sugarcane Biology and Genetic Breeding (Fujian), Ministry of Agriculture, Fujian Agriculture and Forestry University / Sugarcane Research & Development Center, China Agricultural Technology System, Fuzhou 350002, China
  • Received:2014-07-14 Revised:2014-12-09 Online:2015-03-12 Published:2014-12-30
  • Contact: 阙友雄, E-mail: queyouxiong@126.com; 陈如凯, fafu948@126.com

Abstract:

CIPK (calcineurin B-like-interacting protein kinase) is a plant specific class of serine / threonine protein kinases, which plays an important role in plant response to stress, especially relates with the signal transduction for biotic stresses (drought, high salt, ABA). According to the primers designed on the conserved domain of CIPK15 gene from Zea mays, a full-length cDNA sequence of serine/threonine kinase gene termed as ScCIPK was cloned by RT-PCR method from sugarcane (Saccharum Complex). The sequence analysis showed that ScCIPK had a length of 1782 bp containing the open reading frame (ORF, 91–1631 bp), which encoded 513 amino acids residues with two conserved domains (Kc-like superfamily and AMPKA-C-like superfamily). The characters predicted based on the bioinformatics analysis revealed that the ScCIPK gene of sugarcane was a soluble acidic protein, which has two conserved functional domains with the main function for central_intermediary_metabolism, and its protein was located in endoplasmic reticulum (membrane). The mainly secondary structure element was α-helix. Real-time quantitative PCR(RT-qPCR) analysis revealed that the expression of ScCIPK was higher in bud than in other tissues, meanwhile the inducible expression level of ScCIPK was most significantly up-regulated under the ABA stress, 5.3 times higher than that of control, which suggested that ScCIPK most probably involves in sugarcane resistance to drought and osmotic stresses. The results in this study could provide a basis of cloning and functional identification of other members of ScCIPK in sugarcane and promote the use of ScCIPK gene in sugarcane genetic engineering.

Key words: Sugarcane, CIPK, homology cloning, Bioinformatics, Real-time PCR

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