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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (07): 1074-1082.doi: 10.3724/SP.J.1006.2016.01074

• RESEARCH NOTES • Previous Articles     Next Articles

Cloning and Expression Analysis of a Ca2+/H+ Antiporter Gene from Sugarcane

SU Wei-Hua**,LIU Feng**,HUANG Long,SU Ya-Chun,HUANG Ning,LING Hui,WU Qi-Bin,ZHANG Hua,QUE You-Xiong*   

  1. Key Laboratory of Sugarcane Biology and Genetic Breeding (Fujian), Ministry of Agriculture, Fujian Agriculture and Forestry University / SugarcaneResearch & Development Center, China Agricultural Technology System,Fuzhou 350002, China
  • Received:2015-12-07 Revised:2016-03-14 Online:2016-07-12 Published:2016-03-22
  • Contact: 阙友雄, E-mail: queyouxiong@126.com**同等贡献(Contributed equally to this work) E-mail:410946470@qq.com
  • Supported by:

    This study was supported by the China Agriculture Research System (CARS-20), the Special Fund for Agro-Scientific Research in the Public Interest (201503119), and the Program for New Century Excellent Talents in Fujian Province University (JA14095).


CAX (Ca2+/H+ antiporter) is a major category of Ca2+ active transport systems in plant cell membrane. In the present study, using a CAX1 mRNA sequence from Sorghum bicolor (GenBank accession number: XM_002441593) as the probe, the full-length cDNA sequence of sugarcane CAX1 gene was cloned by insilico cloning combined with RT-PCR amplification, and named as ScCAX1 (GenBank accession number: KT799799). Bioinformaticsanalysis showed thatScCAX1 has a length of 784 bp and contains a complete open reading frame with a length of 645 bp, which encodes a 214 amino acid residues of sugarcane CAX1 protein. The ScCAX1 protein with stable acidity and hydrophobia was detected to be located in thylakoid membrane of chloroplasts with no signal peptide. It belongs to a conserved Na_Ca_ex.The mainly secondary structure elementof ScCAX1 protein is alpha helix. Real time quantitative PCR (RT-qPCR)analysis revealed that the expression of ScCAX1 was tissue-specific, with constituent expression in different tissues of sugarcane. The highest expression was observed in leaves while the lowest in stems. Besides, the expression of ScCAX1 gene could be regulated by treatments of PEG, NaCl, SA, ABA, and MeJA. The expression level of this genewas up-regulated by ABA, SA and PEG, with the highest inducible expression level in treatment of 24 hours. The expression level was 5.47 times higher than that of control under 24 h stress of SA , and 3.5 times higher than that of control under 24 h stress of ABA. Under 6 h stress of NaCl, the gene had the highest inducible expression level, which was 2.14 times higher than that of control. This study suggested that ScCAX1 could response to stresses, and its expression may be associated with salt resistance and osmotic tolerance in sugarcane.

Key words: CAX1 gene, nsilico cloning, Bioinformatics, Real-time quantitative PCR

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