Elsevier

Plant Science

Volume 71, Issue 2, 1990, Pages 185-197
Plant Science

Purification and characterization of multiple forms of endochitinase from wheat leaves

https://doi.org/10.1016/0168-9452(90)90008-CGet rights and content

Abstract

The four major forms (I,II,III,VI) of chitinase were purified from leaves of wheat (Triticum aestivum L.) seedlings by affinity chromatography of chitin, and high-performance gel-filtration and ion-exchange chromatography. The isozymes has similar mrs (33, 31.5, 34 and 33 kDa, respectively) as determined by SDS-polyacrylamide gel electrophoresis but widely differing isoelectric points of 5.8, 5.85, 6.7 and 9.1 respectively. Amino acid analyses and double immunodiffusion using specific polyclonal antisera raised in rabbits indicated that the enzymes were closely related. Analysis of the initial products from the digestion of [3H]chitin indicated that all the isozymes were endo in action, with oligomers from the dimer up to at least the decamer being evident in the digests; minor differences were observed between the isozymes in the relative proportions of the oligomeric products. The results are discussed in relation to the release of lignification-elicting chitin oligosaccharides from fungal cell walls during attempted infections of wheat.

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Present address: Department of Biology, The University, Southampton SO9 5NH, U.K.

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