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Related Experiment Videos

Isolation and characterization of a rice gene encoding a basic chitinase.

Q Zhu1, C J Lamb

  • 1Plant Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA 92037.

Molecular & General Genetics : MGG
|April 1, 1991
PubMed
Summary

Researchers identified the rice chitinase RCH10 gene, crucial for plant defense against fungi. This gene is highly expressed in rice roots and induced by fungal elicitors, highlighting its role in plant immunity.

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Area of Science:

  • Plant Molecular Biology
  • Plant Pathology
  • Biochemistry

Background:

  • Chitinase enzymes are vital for plant inducible defenses, targeting fungal cell walls.
  • Understanding plant chitinase genes is key to developing disease resistance strategies.

Purpose of the Study:

  • To isolate and characterize a rice (Oryza sativa L.) chitinase gene.
  • To investigate the gene's structure, expression, and regulation in response to fungal stimuli.

Main Methods:

  • Isolation of a basic chitinase genomic sequence from a rice library using a bean chitinase gene probe.
  • Determination of the complete nucleotide sequence of the RCH10 gene.
  • Gene fusion expression in Escherichia coli and Western blot analysis.
  • Primer extension, Southern blot, and Northern blot analyses for gene expression and genomic organization.

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Main Results:

  • The RCH10 gene encodes a 336-amino acid polypeptide with a signal peptide, hevein domain, and chitinase catalytic domain.
  • RCH10 shares significant nucleotide (63%) and amino acid (75%) identity with dicotyledonous plant chitinases.
  • Transcription start sites and regulatory elements (TATA, CAAT, polyadenylation signal) were identified.
  • Southern blots suggest a chitinase gene family in rice; Northern blots show RCH10 is induced by fungal elicitors.
  • RCH10 transcripts are abundant in roots and less so in stems and leaves.

Conclusions:

  • The RCH10 gene is a functional rice chitinase involved in plant defense mechanisms.
  • Its expression pattern suggests a significant role in root-based resistance to fungal pathogens.
  • The presence of a chitinase gene family indicates a complex defense system in rice.