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Molecular Characterization of an Oomycete-Responsive PR-5 Protein Gene from Zingiber zerumbet

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Abstract

The tropical spice crop ginger (Zingiber officinale Roscoe) is highly susceptible to soft rot disease caused by the necrotrophic oomycete Pythium aphanidermatum (Edson) Fitzp. However, Zingiber zerumbet (L.) Smith, a wild relative of ginger, is resistant to P. aphanidermatum and has been proposed as a potential donor for soft rot resistance to Z. officinale. We identified a member of the pathogenesis-related protein group 5 (PR-5) gene family in Z. zerumbet that is expressed constitutively but upregulated in response to infection by P. aphanidermatum. Expression of this gene was upregulated as early as 1.5 h post inoculation (hpi) with the pathogen, peaked at 6 hpi, declined by 9 hpi, and again peaked at 15 hpi before declining at 48 hpi. A cDNA of this PR-5 gene, designated as ZzPR5, encodes a 226-amino-acid predicted protein with a calculated pI of 5.05. The N terminus of this protein contains a 22-amino-acid signal peptide, suggesting that the protein may show apoplastic accumulation like other acidic PR-5 proteins. Phylogenetic analysis revealed high similarity between ZzPR5 and PR-5 proteins reported from other plant species, especially from other Zingiberales. Molecular modeling of ZzPR5 protein revealed an acidic surface cleft, a feature characteristic of glycoside hydrolases and antifungal PR-5 proteins. In molecular docking studies, a linear polymeric molecule of (1,3)-β-d-glucan, a major constituent of the oomycete cell wall, fitted favorably into the surface cleft of ZzPR5 and interacted with acidic amino acids known to be involved in glucan hydrolysis, suggesting a potential antioomycete activity for ZzPR5 protein. Elucidation of the molecular mechanism of ZzPR5 may provide important insight toward engineering soft rot resistance into the obligatory asexual ginger.

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Abbreviations

hpi:

hours post inoculation

HR:

hypersensitive response

PR:

pathogenesis-related

qRT-PCR:

quantitative real-time polymerase chain reaction

RACE:

rapid amplification of cDNA ends

RMSD:

root mean square deviation

ROS:

reactive oxygen species

TLP:

thaumatin-like protein

ZzPR5:

Zingiber zerumbet pathogenesis-related protein 5

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Acknowledgments

ANR and KAG gratefully acknowledge the Council for Scientific and Industrial Research (CSIR), Government of India for the research fellowships (F. No. 09/716/(0090)/2007/EMR-I to ANR and F. No. 09/716/(0103)/2008/EMR-I to KAG) and GT gratefully acknowledges the Department of Biotechnology (DBT), Government of India for the research grant (Grant No. BT/PR2211/Agr/08/162/2001).

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  1. R. Aswati Nair

    Present address: School of Biotechnology, National Institute of Technology Calicut, NIT Campus P.O., Calicut, 673 601, Kerala, India

Authors and Affiliations

  1. Plant Molecular Biology, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, 695 014, Kerala, India

    R. Aswati Nair, A. G. Kiran & George Thomas

  2. Bioinformatics Division, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, 695 014, Kerala, India

    K. C. Sivakumar

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  1. R. Aswati Nair
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Correspondence to George Thomas.

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Aswati Nair, R., Kiran, A.G., Sivakumar, K.C. et al. Molecular Characterization of an Oomycete-Responsive PR-5 Protein Gene from Zingiber zerumbet . Plant Mol Biol Rep 28, 128–135 (2010). https://doi.org/10.1007/s11105-009-0132-1

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