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Characterization of chitinase activity and gene expression in muskmelon seeds

Published online by Cambridge University Press:  22 February 2007

X. Witmer
Affiliation:
Department of Horticulture, Virginia Tech, Blacksburg, VA 24061-0327, USA
H. Nonogaki
Affiliation:
Department of Crop and Soil Science, Oregon State University, Corvallis, OR 97331-3002, USA
E.P. Beers
Affiliation:
Department of Horticulture, Virginia Tech, Blacksburg, VA 24061-0327, USA
K.J. Bradford
Affiliation:
Department of Vegetable Crops, University of California, Davis, CA 95616-8631, USA
G.E. Welbaum*
Affiliation:
Department of Horticulture, Virginia Tech, Blacksburg, VA 24061-0327, USA
*
*Correspondence Fax: +1 540231 3083, Email: [email protected]

Abstract

Chitinase is often produced in higher plants as a general defence response after wounding or pathogenic attack. Since germinating seeds are exposed to soil pathogens, the activity and expression of chitinase in muskmelon (Cucumis melo L.) seeds was investigated. One acidic and three basic chitinase isoforms were detected, beginning 40 d after anthesis in developing and fully mature seeds. Both acidic and basic chitinase isoforms were found in endosperm tissue during imbibition and after radicle emergence. Basic chitinase isoforms, but not acidic isoforms, were detected in the embryonic axes of imbibed seeds and in seeds before germination, indicating that chitinases are developmentally regulated in specific seed tissues. Two complete cDNAs, Cmchi1 and Cmchi2, were cloned from germinated muskmelon seeds and are predicted to encode chitinases that show 95% identity to a class III chitinase from cucumber (Cucumis sativus L.) and 61% identity to a class II chitinase from soybean (Glycine max L.), respectively. Southern blotting indicated that Cmchi2 was present only once in the muskmelon genome, while Cmchi1 may be present in one or two copies. Cmchi1 and Cmchi2 mRNAs were only detected in radicles of germinating seeds and in roots of mature plants, so additional genes other than Cmchi1 and Cmchi2 must be responsible for the chitinase activity in developing seeds. Salicylic acid and benzothiadiazole stimulated the expression of Cmchi1, but not Cmchi2, after radicle emergence. A putative role for chitinase in muskmelon seeds is defence against fungal pathogens.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2003

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