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Plant Physiol, November 2000, Vol. 124, pp. 1251-1264
Chitinase Genes Responsive to Cold Encode Antifreeze Proteins in
Winter Cereals1
Sansun
Yeh,2
Barbara A.
Moffatt,
Marilyn
Griffith,*
Fei
Xiong,3
Daniel S.C.
Yang,
Steven B.
Wiseman,
Fathey
Sarhan,
Jean
Danyluk,
Yi Qi
Xue,
Choy L.
Hew,4
Amanda
Doherty-Kirby, and
Gilles
Lajoie
Department of Biology, University of Waterloo, Waterloo, Ontario,
Canada N2L 3G1 (S.Y., B.A.M., M.G., S.B.W.); Department of Laboratory
Medicine and Pathobiology, University of Toronto, Toronto, Ontario,
Canada M5G 1L5 (F.X., C.L.H.); Department of Biochemistry, McMaster
University, Hamilton, Ontario, Canada L8N 3Z5 (D.S.C.Y., Y.Q.X.);
Department des Sciences Biologiques, Université du Québec
à Montréal, Montréal, Canada H3C 3P8 (F.S., J.D.);
and Department of Chemistry, University of Waterloo, Waterloo, Ontario,
Canada N2L 3G1 (A.D.-K., G.L.)
Antifreeze proteins similar to two different chitinases accumulate
during cold acclimation in winter rye (Secale cereale). To determine whether these cold-responsive chitinases require post-translational modification to bind to ice, cDNAs coding for two
different full-length chitinases were isolated from a cDNA library
produced from cold-acclimated winter rye leaves. CHT9 is
a 1,193-bp clone that encodes a 31.7-kD class I chitinase and CHT46 is a 998-bp clone that codes for a 24.8-kD class
II chitinase. Chitinase-antifreeze proteins purified from the plant
were similar in mass to the predicted mature products of
CHT9 and CHT46, thus indicating that
there was little chemical modification of the amino acid sequences in
planta. To confirm these results, the mature sequences of
CHT9 and CHT46 were expressed in
Escherichia coli and the products of both cDNAs modified
the growth of ice. Transcripts of both genes accumulated late in cold
acclimation in winter rye. Southern analysis of winter rye genomic DNA
indicated the presence of a small gene family homologous to
CHT46. In hexaploid wheat, CHT46 homologs mapped to the
homeologous group 1 chromosomes and were expressed in response to cold
and drought. We conclude that two novel cold-responsive genes encoding
chitinases with ice-binding activity may have arisen in winter rye and
other cereals through gene duplication.
1
This research was funded by grants from the
Natural Science and Engineering Research Council of Canada (to M.G.,
B.A.M., F.S., and G.L.).
2
Present address: University of Wisconsin,
Department of Botany, 430 Lincoln Drive, Madison, WI 53706.
3
Present address: Amgen, One Amgen Center Drive,
Thousand Oaks, CA 91320.
4
Present address: Department of Biological
Science, National University of Singapore, Singapore.
*
Corresponding author; e-mail griffith{at}uwaterloo.ca; fax
519-746-0614.
© 2000 American Society of Plant Physiologists
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