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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (01): 34-42.doi: 10.3724/SP.J.1006.2013.00034

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Isolation and Functional Analysis of GarCIPK8 Gene from Gossypium aridum

FENG Juan,FAN Xin-Qi,XU Peng,ZHANG Xiang-Gui,SHEN Xin-Lian*   

  1. Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences / Key Laboratory of Cotton and Rapeseed, Ministry of Agriculture, Nanjing 210014, China
  • Received:2012-04-18 Revised:2012-09-05 Online:2013-01-12 Published:2012-11-14
  • Contact: 沈新莲, E-mail: shenxinlian@yahoo.com.cn

Abstract:

CIPK (calcineurin B-like calcium sensor interacting protein kinase) is a typical group of serine / threonine protein kinase targeting calcium sensor calcineurin B-like protein-specific. In a previous study, we obtained the transcriptome of Gossypium aridum with a pooled RNA sample from root and leaf of G. aridum under salt stress. From the transcriptome data, we founded that a CIPK related gene had high level of expression abundance. Real-time fluorescence quantitative PCR analysis showed its relative expression level was up-regulated in root induced by salt stress, indicating this CIPK related gene might be involved in response to salt stress during cotton seedling stage. By in silico cloning and RT-PCR technology, a full-length cDNA encoding a CIPKs homologue (GarCIPK8) was isolated from G. aridum. The deduced GarCIPK8 protein had 449 amino acids with a predicted molecular weight of 51.12 kD and a predicted isoelectric point of 8.13. The kinase N-terminal catalytic domain of GarCIPK8 exhibited a typica1 serine / threonine protein kinase domain. The conserved 24 amino acid motif existed in C-terminal non-kinase regions of GarCIPK8 protein. Alignment of amino acid sequence showed that GarCIPK8 was 73.95% identical to Oryza sativa OsCIPK8 protein. In order to characterize its putative function, the GarCIPK8 gene driven by CaMV35S promoter and stress-specific promoter rd29A respectively were transformed into tobacco by Agrobacterium-mediated transgenic technology. PCR and RT-PCR detection confirmed that GarCIPK8 gene was integrated into tobacco genome and expressed normally. The growth of transgenic plants of T1 generation was observed under salinity and drought stresses, showing that transgenic plants enhance tolerance to both high salt and osmotic stresses.

Key words: Gossypium aridum, GarCIPK8, Salt stress, Drought stress

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