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Plant Physiol, June 2000, Vol. 123, pp. 699-710
Isolation and Characterization of cDNAs Expressed in the
Early Stages of Flavonol-Induced Pollen Germination in
Petunia1
Virginie N.
Guyon,2
James D.
Astwood,3
Ethan C.
Garner,
A. Keith
Dunker, and
Loverine P.
Taylor*
School of Molecular Biosciences, Washington State University,
Pullman, Washington 99163-4234
Petunia (Petunia hybrida) pollen requires flavonols
(Fl) to germinate. Adding kaempferol to Fl-deficient pollen causes
rapid and synchronous germination and tube outgrowth. We exploited this system to identify genes responsive to Fls and to examine the changes
in gene expression that occur during the first 0.5 h of pollen
germination. We used a subtracted library and differential screening to
identify 22 petunia germinating pollen clones. All but two were
expressed exclusively in pollen and half of the clones were rare or low
abundance cDNAs. RNA gel-blot analysis showed that the steady-state
transcript levels of all the clones were increased in response to
kaempferol. The sequences showing the greatest response to kaempferol
encode proteins that have regulatory or signaling functions and include
S/D4, a leucine-rich repeat protein, S/D1, a LIM-domain protein,
and D14, a putative Zn finger protein with a heme-binding site. Eight
of the clones were novel including S/D10, a cDNA only expressed very
late in pollen development and highly up-regulated during the first
0.5 h of germination. The translation product of the S/D3 cDNA
shares some features with a neuropeptide that regulates guidance and
growth in the tips of extending axons. This study confirmed that the
bulk of pollen mRNA accumulates well before germination, but that
specific sequences are transcribed during the earliest moments of
Fl-induced pollen germination.
1
This work was supported by the National Science
Foundation (grant no. IBN-9405361) and by a generous grant from
Pioneer Hi-Bred International (to L.P.T.). Support for sequence
analysis, especially the disorder predictions, was provided by the
National Science Foundation (grant no. 176200 to A.K.D.).
2
Present address: Institut National de la Recherche
Agrono-mique, Laboratoire de biologie des semences, Route de
St. Cyr, 78000 Versailles, France.
3
Present address: Protein Characterization and Safety
Center, Monsanto Company, 700 Chesterfield Parkway North, St. Louis, MO 63198.
*
Corresponding author; e-mail ltaylor{at}wsu.edu; fax 509-335-1907.
© 2000 American Society of Plant Physiologists
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