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A pathogenesis related protein, AhPR10 from peanut: an insight of its mode of antifungal activity

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Abstract

A pathogenesis related protein (AhPR10) is identified from a clone of 6-day old Arachis hypogaea L. (peanut) cDNA library. The clone expressed as a ∼20 kDa protein in E. coli. Nucleotide sequence derived amino acid sequence of the coding region shows its homology with PR10 proteins having Betv1 domain and P loop motif. Recombinant AhPR10 has ribonuclease activity, and antifungal activity against the peanut pathogens Fusarium oxysporum and Rhizoctonia solani. Mutant protein AhPR10-K54N where lys54 is mutated to asn54 loses its ribonuclease and antifungal activities. FITC labeled AhPR10 and AhPR10-K54N are internalized by hyphae of F. oxysporum and R. solani but the later protein does not inhibit the fungal growth. This suggests that the ribonuclease function of AhPR10 is essential for its antifungal activity. Energy and temperature dependent internalization of AhPR10 into sensitive fungal hyphae indicate that internalization of the protein occurs through active uptake.

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Abbreviations

AhPR10:

Arachis hypogaea pathogenesis related 10 protein

AhPR10-F148S:

AhPR10 mutant protein where phenylalanine148 is replaced by serine

AhPR10-H150Q:

AhPR10 mutant protein where histidine150 is replaced by glutamine

AhPR10-K54N:

AhPR10 mutant protein where lysine54 is replaced by asparagine

FITC:

Fluorescein isothiocyanate

IC50 :

Inhibitory concentration causing 50% growth inhibition

IPTG:

Isopropyl thio-β-galactoside

NaN3 :

Sodium azide

PBS:

Phosphate buffered saline, pH 7.2

PCR:

Polymerase chain reaction

PI:

Propidium iodide

PMSF:

Phenyl methyl sulfonyl fluoride

PR:

Pathogenesis related

SDS-PAGE:

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

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Acknowledgments

The authors gratefully acknowledge the Director of the Institute of Genomics and Integrative Biology for providing the infrastructural facilities to carry out this work. Authors also thank Dr. Pratibha Sharma of Mycology and Plant Pathology Division, IARI, New Delhi, India for providing the fungal pathogens. PC is the recipient of Research Fellowship from CSIR (India).

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Authors and Affiliations

  1. Comparative Genomics Unit, Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi, 110 007, India

    Pooja Chadha & Rakha H. Das

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  1. Pooja Chadha
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  2. Rakha H. Das
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Correspondence to Rakha H. Das.

Additional information

The nucleotide sequence of AhPR10 reported in this paper is submitted to NCBI Nucleotide Sequence Database under the Accession number AY726607.

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Chadha, P., Das, R.H. A pathogenesis related protein, AhPR10 from peanut: an insight of its mode of antifungal activity. Planta 225, 213–222 (2006). https://doi.org/10.1007/s00425-006-0344-7

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