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A class I chitinase from soybean seed coat.
1Agriculture and Agri-Food Canada, 1391 Sandford Street, London, Ontario, Canada N5V 4T3. gijzenm@em.agr.ca
Journal of Experimental Botany
|November 16, 2001
Summary
Soybean seed coats contain a chitinase enzyme, a defense protein, that is highly expressed during seed development and in response to pathogens. This chitinase has a hevein domain and is encoded by a single-copy gene.
Area of Science:
- Plant molecular biology
- Biochemistry
- Genetics
Background:
- Soybean seed coat proteins exhibit diverse functions.
- Chitin-binding proteins, including chitinases, play roles in plant defense.
Purpose of the Study:
- To isolate and characterize a chitinase enzyme from soybean seed coats.
- To investigate the gene structure, expression, and function of this soybean chitinase.
Main Methods:
- Protein fractionation (isoelectric focusing, SDS-PAGE)
- Peptide microsequencing
- Chitin affinity chromatography
- Zymogram assay
- cDNA and genomic cloning
- RNA and DNA blot analysis
- Gene sequencing
Main Results:
- An abundant 32 kDa soybean protein with a hevein domain and chitinase activity was identified.
- The corresponding cDNA and gene were isolated; the gene contains two introns and is a single or low copy gene.
- Chitinase expression is high in late seed development, particularly in the seed coat, and also occurs in other tissues and upon pathogen infection.
Conclusions:
- Soybean seed coats possess a defense-related chitinase with a hevein domain.
- This chitinase is encoded by a single-copy gene and its expression is developmentally regulated and inducible by pathogens.
- The findings highlight the prevalence of defense proteins in soybean seed coats.