xidation is stepwise—removal of the first

oxygen generates an intermediate monoepoxide

(antheraxanthin)(A). De-epoxidation is also induced by
a low pH in the lumen, which is a normal consequence
of electron transport under high light conditions.
The reaction is reversed in the dark as zeaxanthin is
again enzymatically converted back to violaxanthin.
Xanthophyll cycle activity can be assessed by measuring
the de-epoxidation state (DEPS) of the cycle pool
which is a measure of the concentration of zeaxanthin
(Z) and antheraxanthin (A) relative to the total pool
(V + A + Z) (Equation 14.1).
DEPS = Z + 1
2A/V + A + Z (14.1)
Although it has been established that the xanthophyll cycle plays a key role in photoprotection of
the chloroplast through nonphotochemical quenching (NPQ; Box 13.1), the precise molecular mechanism
246 Chapter 14 / Acclimation to Environmental Stress
is still disputed. Through the generation of an Arabidopsis thaliana mutant named npq4, one model suggests
that the zeaxanthin produced by the xanthophyll cycle
may be bound to a specific PSII polypeptide encoded
by the PsbS gene. These observations form the basis
for the hypothesis that the xanthophyll cycle acts as a
reversible, molecular switch to regulate the capacity for
NPQ and the safe dissipation of excess absorbed. Under
low light, LHCII has abundant violaxanthin (V) whose
absorbed excitation energy is rapidly transferred to the
reaction center to excite P680 to P680* (Figure 14.4B).
However, exposure to high light rapidly converts to
violaxanthin to zeaxanthin within LHCII. Since Z can
not transfer its excitation energy to chlorophyll, Z in
the excited state decays to its ground state by losing its
excitation energy as heat. The molecular switch model
assumes that zeaxanthin (Z) absorbs light energy directly
and that NPQ is a consequence of the decay of excited
state Z to its ground state with the concomitant loss of
heat. Thus, zeaxanthin decreases the efficiency of energy
transfer to PSII reaction centers by acting as a direct
quencher of energy. Support for the molecular switch
model is still equivocal since the presence of PsbS still
has not been detected in the crystal structure of PSII,
and furthermore, NPQ can occur in the absence of Z.
An alternative model for the regulation of NPQ
suggests that the conversion of V to Z by the xanthophyll cycle regulates the aggregation state of LCHII.
In the aggregated state, the efficiency of energy transfer from LHCII to PSII reaction centers is drastically
decreased. In this model, aggregated LHCII acts as the
energy quencher which protects PSII reaction centers
from excess excitation. This suggests that Z is indirectly
involved in quenching energy by affecting the physical
structure of LHCII. Regardless of which model is correct, it appears that the xanthophyll cycle is a ubiquitous
process for protecting the chloroplast against potentially damaging effects of excess light. However, it is
interesting to note that the xanthophyll cycle is absent
in cyanobacteria. The regulation of NPQ by the xanthophyll cycle continues to be an intensive and exciting
area of photosynthesis research.
What are the functional consequences of a stimulation of NPQ by high light? Irrespective of the
mechanism underlying photoprotection through NPQ,
the functional consequence of NPQ is a decrease in
the efficiency of energy transfer to PSII reaction centres. Because less energy is transferred to PSII reaction
centers per photon absorbed by LCHII under such
conditions, the efficiency of PSII photochemistry (P680
+ energy → P680+ + e), will decrease per photon
absorbed. Thus, the light-inducible xanthophyll cycle
protects PSII reaction centers by dissipating excess excitation nonphotochemically as heat.
When plants are subjected to excess light,
photoinhibition will occur (Chapter 13) which can
be measured as a decrease in Fv/Fm as a function
of time under high light. However, concomitantly,
exposure to light also stimulates the de-epoxidation
of the xanthophyll cycle (DEPS) and NPQ over
time (Figure 14.5A, left panel). However, when the
plants are allowed to recover from the photoinhibition
by removal from the high light condition, Fv/Fm
and DEPS rapidly recover to their original values
prior to the photoinhibition treatment (Figure 14.5A,
right panel). Thus, the responses of Fv/Fm
500
Irradiance (mol m2 s1)
0 750 250
30
20
Rate of Photosynthesis
(mol O2 evolved m2 s1)
10
Photoinhibition
Time (min)
Recovery
Time (min)
0 40 80 40 80
0.8
0.6
0.4
0.2
Fv/Fm ( , )
DEPS ( , )
A.
B.
FIGURE 14.5 A schematic graph illustrating photoprotection and dynamic photoinhibition. (A) The effects of
time of exposure to photoinhibition and time of recovery from this photoinhibition on Fv/Fm (black curve) and
DEPS (red curve). The panel on the left illustrates that
the time-dependent decrease in Fv/Fm is almost a mirror image of time-dependent increase in DEPS due to
exposure to high light. The panel on the right illustrates
that the time-dependent recovery of Fv/Fm (black broken line) is almost a mirror image of the time-dependent
decrease in DEPS (red broken line). Such mirror image
responses are characteristic of dynamic photoinhibition.
In contrast to dynamic photoinhibition, recovery from
chronic photoinhibition is much slower (blue broken
line). (Adapted from Pocock, T., D. Koziak, D., Rosso,
N. P. A. Huner. 2007. Journal of Phycology 43:924–936.)
(B) The effects of exposure to either dynamic (red
line) or chronic photoinhibition (broken red line) on
the light response curves for O2 evolution. The black
line represents the light response curve for control,
non-photoinhibited plants. (Adapted from Osmond, C.
B. 1994. In: N. R. Baker, J. R. Bowyer (eds.), Photoinhibition of Photosynthesis: From Molecular Mechanisms to the
Field, pp. 1–24. Oxford: Bios Scientific Publishers.)
14.2 Acclimation is Initiated by Rapid, Short-Term Responses 247
and DEPS to photoinhibition and recovery time
appear to be mirror images of one another. The
rapidly reversible inhibition of PSII reaction centers
that is usually a consequence of an increase in thermal
energy dissipation through NPQ is defined as dynamic
photoinhibition. This reflects photoprotection. In
contrast, photodamage or chronic photoinhibition
is defined as the slowly reversible inhibition of PSII
reaction centers that is usually a consequence of damage
to the D1 reaction center polypeptide. Thus, the time
required for the recovery of Fv/Fm from chronic
photoinhibition is much longer than that from dynamic
photoinhibition. Photodamage is only slowly reversible
because of its dependence on protein synthesis for the
D1 repair cycle (Chapter 13). The difference between
dynamic and chronic photoinhibition can also be seen
at the level of O2 evolution (Figure 14.5B). Dynamic
photoinhibition usually results in a rapidly reversible
decrease in photosynthetic efficiency (Chapter 13)
measured as the maximum slope of the CO2 light
response curve but not a decrease in photosynthetic
capacity measured as the maximum light saturated rate
of photosynthesis. In contrast, chronic photoinhibition
usually results in a decrease in both photosynthetic
efficiency and photosynthetic capacity which recover
very slowly (Figure 14.5B). Clearly, both photodamage
and photoprotection may lead to photoinhibition
of photosynthesis as reflected in a decrease in
photosynthetic efficiency, albeit for different reasons.
The former causes a reduction in photosynthetic
efficiency due to an alteration in the antenna that
reduces the efficiency of resonance energy transfer to
the PSII reaction center. The latter is due to damage to
the reaction center that reduces the efficiency of charge
separation rather than energy transfer in the antenna.
14.2.3 OSMOTIC ADJUSTMENT IS
A RESPONSE TO WATER STRESS
A pronounced response to water stress in many plants
is a decrease in osmotic potential resulting from an
accumulation of solutes (see Chapter 1 and 2). This
process is known as osmotic adjustment. While some
increase in solute concentration is expected as a result
of dehydration and decreasing cell volume, osmotic
adjustment refers specifically to a net increase in solute
concentration due to metabolic processes triggered by
stress. Osmotic adjustment generates a more negative
leaf water potential, thereby helping to maintain water
movement into the leaf and, consequently, leaf turgor.
Solutes accumulate during osmotic adjustment and
the decreases in osmotic potential due to osmotic
adjustment are relatively small, less than 1.0 MPa. Nevertheless, the role of solutes in maintaining turgor at
relatively low water potentials represents a significant
form of acclimation to water stress. Osmotic adjustment
may also play an important role in helping partially
wilted leaves to regain turgor once the water supply
recovers. By helping to maintain leaf turgor, osmotic
adjustment also enables plants to keep their stomata
open and continue taking up CO2 for photosynthesis under conditions of moderate water stress. Solutes
implicated in osmotic adjustment include a range of
inorganic ions (especially K+), sugars, and amino acids
(Figure 14.6). One amino acid that appears to be particularly sensitive to stress is proline. A large number of
plants synthesize proline from glutamine in the leaves.
The role of proline is demonstrated by experiments
with tomato cells in culture. Cells subjected to water
(osmotic) stress by exposure to hyperosmotic concentrations of polyethylene glycol (PEG) responded with an
initial loss of turgor and rapid accumulation of proline.
As proline accumulation continued, however, turgor
gradually recovered. Sorbitol, a sugar alcohol, and
betaine (N,N,N-trimethyl glycine) are other common
accumulated solutes. Most chemicals associated with
osmotic adjustment share the property that they do not
significantly interfere with normal metabolic processes.
Such chemicals are called compatible solutes.
Although osmotic adjustment appears to be a general response to water stress, not all species are capable
of adjusting their solute concentrations. Sugarbeet (Beta
vulgaris), on the one hand, synthesizes large quantities of
betaine and is known as an osmotic adjuster. Osmotic
adjustment in cowpea (Vigna unguiculata), on the other
hand, is minimal and cowpea is known as an osmotic
nonadjuster. Cowpea instead has very sensitive stomata
an