Isolation and Characterization of a Chitinase and its cDNA Clone from Rice

  • C. S. Anuratha
  • Jenq-Kuen Huang
  • Arunabala Pingali
  • S. Muthukrishnan
Article

Abstract

A 35 kD chitinase has been purified to apparent homogeneity from extracts of rice bran of cv New Bonnet by ammonium sulfate fractionation, chitin affinity chromatography, cation exchange chromatography on carboxymethyl cellulose and gel filtration. The purified enzyme has an isoelectric point of 8.8. The enzyme inhibited the growth of Rhizoctonia solani (the sheath blight pathogen), Trichoderma viride, T. harzianum, Fusarium graminaerum and F. culmorum in vitro.

A cDNA clone for chitinase was isolated from a developing rice seed cDNA library by probing with a barley chitinase cDNA probe. The nucleotide sequence of this 654 bp clone was determined, it contains an open reading frame of 519 nucleotides. The protein product encoded by this clone is homologous to chitinases from tobacco, bean and barley. Southern blot analysis of rice genomic DNA with this probe revealed that chitinases are encoded by a small multi-gene family in the rice genome.

Key words

chitinase characterization antifungal properties cDNA clone chitinase genes rice 

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Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • C. S. Anuratha
    • 1
  • Jenq-Kuen Huang
    • 2
  • Arunabala Pingali
    • 1
  • S. Muthukrishnan
    • 1
  1. 1.Department of BiochemistryKansas State UniversityManhattanUSA
  2. 2.Department of ChemistryWestern Illinois UniversityMacombUSA

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