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Plant Physiol, February 2002, Vol. 128, pp. 534-543
Temporal and Spatial Expression of a Polygalacturonase during
Leaf and Flower Abscission in Oilseed Rape and
Arabidopsis1
Zinnia Haydé
González-Carranza,2
Catherine
Ann
Whitelaw,3
Ranjan
Swarup, and
Jeremy Alan
Roberts2*
Plant Science Division, School of Biosciences, University of
Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire LE12
5RD, United Kingdom
During leaf abscission in oilseed rape (Brassica
napus), cell wall degradation is brought about by the action of
several hydrolytic enzymes. One of these is thought to be
polygalacturonase (PG). Degenerate primers were used to isolate a PG
cDNA fragment by reverse transcriptase-polymerase chain reaction
from RNA extracted from ethylene-promoted leaf abscission zones (AZs),
and in turn a full-length clone (CAW471) from an oilseed rape AZ cDNA
library. The highest homology of this cDNA (82%) was to an Arabidopsis sequence that was predicted to encode a PG protein. Analysis of expression revealed that CAW471 mRNA accumulated in the AZ of leaves
and reached a peak 24 h after ethylene treatment.
Ethylene-promoted leaf abscission in oilseed rape was not apparent
until 42 h after exposure to the gas, reaching 50% at 48 h
and 100% by 56 h. In floral organ abscission, expression of
CAW471 correlated with cell separation. Genomic libraries from oilseed
rape and Arabidopsis were screened with CAW471 and the respective
genomic clones PGAZBRAN and PGAZAT
isolated. Characterization of these PG genes revealed that they had
substantial homology within both the coding regions and in the
5'-upstream sequences. Fusion of a 1,476-bp 5'-upstream sequence of
PGAZAT to  -glucuronidase or green fluorescent protein and transformation of Arabidopsis revealed that this fragment was
sufficient to drive expression of these reporter genes in the AZs at
the base of the anther filaments, petals, and sepals.
1
This work was supported by Consejo Nacional de
Ciencia y Tecnología (México; fellowship no. 94459/110150
to Z.H.G.-C.) and by the Biotechnology and Biological Science Research Council.
2
Z.H.G.-C. and J.A.R. have set up a Web site
focused on abscission and dehiscence, located at
http://arabidopsis.info/abscission.
3
Present address: The Institute for Genomic Research,
9712 Medical Center Drive, Rockville, MD 20850.
*
Corresponding author; e-mail jeremy.roberts{at}nottingham.ac.uk;
fax 44-115951-6334.
© 2002 American Society of Plant Physiologists
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