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Interaction of Cryptochrome 1, Phytochrome, and Ion Fluxes in
Blue-Light-Induced Shrinking of Arabidopsis
Hypocotyl
Protoplasts1
Xiaojing Wang2 and
Moritoshi Iino*
Botanical Gardens, Faculty of Science, Osaka City University,
Kisaichi, Katano-shi, Osaka 576, Japan
Protoplasts
isolated from red-light-adapted Arabidopsis hypocotyls and incubated
under red light exhibited rapid and transient shrinking within a period
of 20 min in response to a blue-light pulse and following the onset of
continuous blue light. Long-persisting shrinkage was also observed
during continuous stimulation. Protoplasts from a hy4
mutant and the phytochrome-deficient
phyA/phyB double mutant of Arabidopsis
showed little response, whereas those from phyA and
phyB mutants showed a partial response. It is concluded that the shrinking response itself is mediated by the
HY4 gene product, cryptochrome 1, whereas the blue-light
responsiveness is strictly controlled by phytochromes A and B, with a
greater contribution by phytochrome B. It is shown further that the
far-red-absorbing form of phytochrome (Pfr) was not required during or
after, but was required before blue-light perception. Furthermore, a
component that directly determines the blue-light responsiveness was
generated by Pfr after a lag of 15 min over a 15-min period and decayed with similar kinetics after removal of Pfr by far-red light. The anion-channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoic acid prevented the shrinking response. This result, together with those
in the literature and the kinetic features of shrinking, suggests that
anion channels are activated first, and outward-rectifying cation
channels are subsequently activated, resulting in continued net
effluxes of Cl and K+. The postshrinking
volume recovery is achieved by K+ and Cl
influxes, with contribution by the proton motive force. External Ca2+ has no role in shrinking and the recovery. The gradual
swelling of protoplasts that prevails under background red light is
shown to be a phytochrome-mediated response in which phytochrome A
contributes more than phytochrome B.
1
This work was supported by a Monbusho's
grant-in-aid for the Japan Society for Promotion of Science (JSPS)
fellows. X.W. was the recipient of a JSPS postdoctoral fellowship.
2
Present address: Department of Biology, South
China Normal University, Guangzhou 510631, China.
*
Corresponding author; e-mail iino{at}sci.osaka-cu.ac.jp; fax
81-720-91-7199.
Plant Physiol. (1998) 117: 1265-1279
Copyright Clearance Center: 0032-0889/98/117//15
© 1998 American Society of Plant Physiologists
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