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Plant Physiol, October 2001, Vol. 127, pp. 473-485
Dominant Negative Guard Cell K+ Channel Mutants
Reduce Inward-Rectifying K+ Currents and Light-Induced
Stomatal Opening in Arabidopsis1
June M.
Kwak,2
Yoshiyuki
Murata,2
Victor M.
Baizabal-Aguirre,3
Jennifer
Merrill,
Michele
Wang,
Andrea
Kemper,
Scott D.
Hawke,
Gary
Tallman, and
Julian I.
Schroeder*
Division of Biology, Cell and Developmental Biology Section, and
Center for Molecular Genetics, University of California, San Diego,
9500 Gilman Drive, La Jolla, California 92093-0116 (J.M.K, Y.M.,
V.M.B.-A., J.M., M.W., J.I.S.); and Department of Biology, Willamette
University, 900 State Street, Salem, Oregon 92037-3931 (A.K., S.D.H.,
G.T.)
Inward-rectifying potassium (K+in) channels
in guard cells have been suggested to provide a pathway for
K+ uptake into guard cells during stomatal opening. To test
the proposed role of guard cell K+in channels
in light-induced stomatal opening, transgenic Arabidopsis plants were
generated that expressed dominant negative point mutations in the
K+in channel subunit KAT1. Patch-clamp analyses with transgenic guard cells from independent lines showed that K+in current magnitudes were reduced by
approximately 75% compared with vector-transformed controls at  180
mV, which resulted in reduction in light-induced stomatal opening by
38% to 45% compared with vector-transformed controls. Analyses of intracellular K+ content using both sodium
hexanitrocobaltate (III) and elemental x-ray microanalyses showed that
light-induced K+ uptake was also significantly reduced in
guard cells of K+in channel depressor lines.
These findings support the model that K+in
channels contribute to K+ uptake during light-induced
stomatal opening. Furthermore, transpirational water loss from leaves
was reduced in the K+in channel depressor
lines. Comparisons of guard cell K+in current
magnitudes among four different transgenic lines with different
K+in current magnitudes show the range of
activities of K+in channels required for guard
cell K+ uptake during light-induced stomatal opening.
1
This work was supported by the Department of
Energy (grant no. De-FG03-94-ER20148 to J.I.S.), by the National
Science Foundation (grant nos. MCB-9506191 and MCB-00-77791 to
J.I.S., grant no. MCB-9900525 to G.T., and REU supplement to
J.I.S.), by the Human Frontier Science Program Organization (fellowship
to J.M.K.), by the Pew Foundation (fellowship to V.M.B.-A.), and by the
Ministry of Education, Science, Sports and Culture of Japan (fellowship to Y.M.).
2
These authors contributed equally to this work.
3
Present Address: Centro Multidisciplinario de Estudios
en Biotecnologia, Facultad de Medicina Veterinaria y Zootecnia,
Universidad Michoacana de San Nicolas de Hidalgo, kilometro 9.5 Carretera Morelia-Zinapecuaro, La Palma, Tarimbaro, Michoacan, Mexico.
*
Corresponding author; e-mail julian{at}biomail.ucsd.edu; fax
858-534-7108.
© 2001 American Society of Plant Physiologists
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