CDPK1 from ginger promotes salinity and drought stress tolerance without yield penalty by improving growth and photosynthesis in Nicotiana tabacum

PLoS One. 2013 Oct 23;8(10):e76392. doi: 10.1371/journal.pone.0076392. eCollection 2013.

Abstract

In plants, transient changes in calcium concentrations of cytosol have been observed during stress conditions like high salt, drought, extreme temperature and mechanical disturbances. Calcium-dependent protein kinases (CDPKs) play important roles in relaying these calcium signatures into downstream effects. In this study, a stress-responsive CDPK gene, ZoCDPK1 was isolated from a stress cDNA generated from ginger using rapid amplification of cDNA ends (RLM-RACE) - PCR technique and characterized its role in stress tolerance. An important aspect seen during the analysis of the deduced protein is a rare coupling between the presence of a nuclear localization sequence in the junction domain and consensus sequence in the EF-hand loops of calmodulin-like domain. ZoCDPK1 is abundantly expressed in rhizome and is rapidly induced by high-salt stress, drought, and jasmonic acid treatment but not by low temperature stress or abscissic acid treatment. The sub-cellular localization of ZoCDPK1-GFP fusion protein was studied in transgenic tobacco epidermal cells using confocal laser scanning microscopy. Over-expression of ginger CDPK1 gene in tobacco conferred tolerance to salinity and drought stress as reflected by the high percentage of seed germination, higher relative water content, expression of stress responsive genes, higher leaf chlorophyll content, increased photosynthetic efficiency and other photosynthetic parameters. In addition, transgenic tobacco subjected to salinity/drought stress exhibited 50% more growth during stress conditions as compared to wild type plant during normal conditions. T3 transgenic plants are able to grow to maturity, flowers early and set viable seeds under continuous salinity or drought stress without yield penalty. The ZoCDPK1 up-regulated the expression levels of stress-related genes RD21A and ERD1 in tobacco plants. These results suggest that ZoCDPK1 functions in the positive regulation of the signaling pathways that are involved in the response to salinity and drought stress in ginger and it is likely operating in a DRE/CRT independent manner.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cluster Analysis
  • DNA Primers / genetics
  • Droughts
  • Gene Expression Regulation, Enzymologic / genetics*
  • Gene Transfer Techniques
  • Microscopy, Confocal
  • Molecular Sequence Data
  • Nicotiana / genetics
  • Nicotiana / growth & development*
  • Nucleic Acid Amplification Techniques
  • Phylogeny
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / physiology
  • Protein Kinases / genetics*
  • Protein Kinases / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Salinity
  • Sequence Analysis, DNA
  • Stress, Physiological / genetics*
  • Zingiber officinale / chemistry*

Substances

  • DNA Primers
  • Protein Kinases
  • calcium-dependent protein kinase

Grants and funding

The current work was supported by Kerala State Council for Science, Technology and Environment [grant number: 1418/C3/06/KSCSTE]. P. J. Vivek received fellowship from University Grant Commission (UGC), New Delhi, India [F.No.10-2(5)/2005(ii)-E.U.II]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.