movement?

These questions have been approached

with a variety of techniques, including histochemical and radiochemical methods, and scanning electron
microscopy coupled with X-ray analysis. The results
show that leaf movement in all nyctinastic plants studied thus far is associated with a massive redistribution of
potassium ion (K+) between the symplast and apoplast
in both the extensor and flexor regions of the pulvinus
(Figure 23.20). Swollen extensor cells are characterized
by high protoplasmic K+ and low apoplastic K+. In
Phaseolus vulgaris, fully 30 percent of the osmotic potential change can be accounted for by K + movement. The
charge carried by K+ is compensated primarily by chloride and possibly small organic anions such as malate
and citrate.
Beyond the central role of K+ flux in nastic movements, it is difficult to pin down the specific sequence
of molecular and biophysical events in the signal chain.
Patch-clamp experiments with isolated Samanea motor
cell protoplasts have established that K+ exchange across
the plasma membrane occurs through K+ channels and
that these channels can be regulated by changing the
membrane polarity. Depolarization of the membrane
opens the channels and allows K+ to move out of the
cell down its electrochemical gradient. It has also been
established that there are pH gradients across the plasma
membranes of motor cells. In the case of open Samanea
pulvina, the pH of the apoplast is about 5.5 in the
flexor region and 6.2 in the extensor region. The cytoplasmic pH is approximately neutral in both regions.
Time (min)
Upon a light/dark transition (leading to closure), the FIGURE 23.20 Changes in K+ activity in the apoplast of

pH gradient in the extensor region dissipates while

in the flexor region the gradient increases. Although
quantitative relationships between H+ flux and K+ flux
have yet to be tested, H+ extrusion could contribute
to the electrochemical gradient necessary to drive K+

uptake. Any observed changes in membrane potential

are undoubtedly the consequence, not the cause, of
the
cross-membrane traffic in osmotically active ions.
The prominent roles of K+ channels and H+
pumps have been incorporated into the current model
for motor cell movement. A simplified version of the
model is shown in Figure 23.21. In this model, the
light signal activates phytochrome (or cryptochrome), graphs. (Adapted from Lowen, C. Z., R. L. Satter.
which accelerates inositol phospholipid turnover. 1989.
Recent experiments have shown that light that Light promoted changes in the apoplastic K+ activity
in the Samanea samanan pulvinus. Planta 179:412–427,
stimulates opening of Samanea pulvini also decreases
Figure 1. Copyright Springer-Verlag.)
the level of phosphotidylinositol 4,5-bisphosphate
(PIP2) and increases the level of the second
messenger
inositol 1,4,5-triphosphate (IP3). There is a transient
stimulation of diacylglycerol (DAG). These changes
are qualitatively similar but quantitatively smaller
than those normally detected in animal systems. The
assays, however, involved whole pulvini, which
contain
 vascular, collenchyma, and epidermal tissues as well
as

the motor cells. The changes could be appropriately

greater if restricted to the smaller population of motor
cells. If inositol phospholipid metabolism functions in
plants as it does in animals, DAG would be expected
to activate a protein-kinase C (or its plant equivalent)
to phosphorylate certain proteins. IP 3 would be
expected to release free calcium—exogenous IP3 does
liberate calcium—although from which compartment
is not known. Both the phosphorylated protein and/or
transient increases in free calcium stimulate proton
extrusion by activating the proton pump. The
resulting
electrochemical gradient energizes the uptake of K+
and other ions, which in turn stimulates the osmotic
0
10
20
K+ Activity (mM)
30
40
50
60
70
80
0
10
20
30
0 20 40 60 80 100
extensor (upper curve) and flexor (lower curve) cells
during closure of Samanea leaflets. Loss of K+ from the
protoplasts is followed by loss of water and turgor. Closure and opening were stimulated by dark and white
light periods as indicated by the bar between the two

23.3 Nastic Movements 409

+


]] Cytosol
K+ C1–
[IP3 [Ca2+

Plasma
membrane
P
r
?
P
fr PIP2
[DAG]

Ca2+
Protein + ATP Protein ~ P
h
H+
FIGURE 23.21 A proposed model for the interaction of phytochrome, biological
clocks, and the inositol triphosphate system in leaf movements of nyctinastic plants.
Light, mediated by phytochrome and modulated by the endogenous clock, accelerates inositol phospholipid turnover
and increases the level of the second messengers
inositol-1,4,5-triphosphate (IP3) and diacylglycerol (DAG). The second messengers
stimulate a release of Ca2+ into the cytosol and phosphorylation of various proteins
which in turn stimulate the extrusion of protons from the cell. K + diffuses into the
cell in response to the proton motive force. An active transport pump extrudes Ca2+
as an aid to restoring Ca2+ homeostasis.
uptake of water and motor cell swelling. The presence
of a calcium pump that extrudes Ca2+ would help to
ensure the restoration of Ca2+ homeostasis.
Many details of this model remain to be described,
especially the function of the inositol phospholipid cycle
in plants. Still, plant cells are known to contain virtually
all the required components and the model is consistent
with what has been observed in pulvini thus far. Significant advances are to be expected in the future, especially
now that patch-clamp techniques—long a mainstay of
electrophysiology research for animal cells—can be
applied to plant cell protoplasts. This state-of-the-art
technique has been in use for plant cells only since
about 1984, but has proven invaluable for the study of
ion channels.
23.3.3 SEISMONASTY IS A RESPONSE TO
MECHANICAL STIMULATION
A limited number of leguminous plants that possess
pulvini and exhibit nyctinastic movements also exhibit a
response to mechanical stimulation. This phenomenon
is known as seismonasty. Since seismonastic plants
respond to touch, they are sometimes considered
thigmonastic. However, seismonastic plants respond to
a wider variety of stimuli including shaking or wind,
falling raindrops, wounding by cutting, and intense heat
or burning.
The best known