J Agro Crop Sci (2015) ISSN 0931-2250

CHILLING/FREEZING STRESS

Photosynthetic Apparatus Responses to Short-term

low-temperature Flooding May Contribute to Freezing
Tolerance Changes in Forage Grasses
B. Jurczyk, M. Rapacz & T. Krez pski

w, Krako
Department of Plant Physiology, University of Agriculture in Krako
w, Poland

Keywords Abstract
Chlorophyll a fluorescence; cold acclimation;
freezing tolerance Winter conditions are subjected to rapid climate changes. Increased precipitation
and snow melting during warmer winters may result in low-temperature flood-
Correspondence ing. These factors probably affect plant overwintering strategies. This study inves-
B. Jurczyk tigated the relationships between the photosynthetic adjustment and freezing
Department of Plant Physiology
tolerance following a short-term low-temperature flooding in two forage grasses

w
University of Agriculture in Krako
(Lolium perenne and Festuca pratensis). The effect of flooding on the photosyn-
Podłuz_ na 3
30-239 Krako
w thetic apparatus acclimation to cold was determined using chlorophyll a fluores-
Poland cence measurements. Freezing tolerance was estimated using an electrolyte
Tel.: +12 425 33 01 leakage test. It has been shown that genotypes activating non-photochemical
Fax: +12 425 33 20 mechanisms of photosynthetic acclimation to cold during low-temperature
Email: b.jurczyk@ur.krakow.pl flooding may show increased freezing tolerance. Freezing tolerance reduction
observed in Festuca pratensis was probably connected with decreased photochem-
Accepted July 14, 2014
ical activity and photoinhibition of photosynthesis. It can be concluded that pho-
doi:10.1111/jac.12089 tosynthetic apparatus response to low-temperature flooding may contribute to
changes in the freezing tolerance. The direction of the changes is associated with
different photosynthetic apparatus performance.

serve as a signal triggering both the increase in freezing tol-

Introduction
Intense winter precipitation events seem to become
increasingly common, and they may have serious flooding-
related consequences (Osborn and Hulme 2002). The most
important effects of the soil flooding are a reduction of soil
nutrient uptake and a decrease in a plant respiratory
metabolism (Dat et al. 2004). The reduced photosynthetic
rate observed during soil flooding has been attributed to
stomatal closure and leaf chlorophyll decrease (Bradford
1983) and also to a reduced sink demand (Wample and
Thornton 1984).
During winter, the photosynthetic enzymes are inhibited
by low temperature, but light absorption is continued
€
(Oquist and H€uner 2003, Adams et al. 2004). Light energy,
absorbed in the light processes, may exceed the energy con-
sumption in the dark processes, which can result in an
over-reduction of photosystem II and photoinhibition of
photosynthesis. Increased reduction of photosystem II may
erance and the photoinhibition avoidance. Thus, the toler-
ance to cold-induced photoinhibition of photosynthesis
seems to be related to freezing tolerance, which is a con-
sequence of common mechanisms of acclimation to both
stresses (H€uner et al. 1996).
Higher plants exhibit two main strategies of the photo-
synthetic apparatus acclimation to cold, based on photo-
chemical and non-photochemical mechanisms (H€ uner
et al. 1993). The photochemical mechanism depends on an
increased capacity of photosynthesis, due to intensified car-
bon assimilation and carbon metabolism. The second strat-
egy, based on a non-photochemical mechanism, involves
the dissipation of excess energy as heat (H€ uner et al. 1993,
M€ uller et al. 2001). Although the photochemical mecha-
nism has been developed rather by overwintering herba-
ceous plants and the non-photochemical mechanism is
generally attributed to evergreen trees and shrubs, both
mechanisms may be activated simultaneously and their

© 2014 Blackwell Verlag GmbH, 201 (2015) 49–56 49

Jurczyk et al.
expression level depends on a plant genotype (Rapacz et al.
2004, Humphreys et al. 2007) and environmental factors
(Adams et al. 2002).
Acclimation of the photosynthetic apparatus to low-tem-
perature conditions is very important for perennial forage
grasses as it facilitates the development of winter hardiness
and freezing tolerance (Rapacz et al. 2004). Plants impaired
in photosynthetic acclimation to cold are also unable to
increase their freezing tolerance during cold acclimation
(CA) (Rapacz et al. 2007). These relationships make the
perennial forage grasses an interesting model system for
studying the mechanisms of the photosynthetic acclimation
to cold.
Although the effect of low-temperature flooding on frost
tolerance is not fully understood (Rakitina 1977, Andrews
and Pomeroy 1981, Jurczyk et al. 2013), it is assumed that
anaerobic processes initiated during flooding promote sur-
vival at the time of subsequent ice encasement (Andrews
and Pomeroy 1981). It has been shown (Jurczyk et al.
2013) that low-temperature flooding of the plant roots and
crowns may enhance CA efficiency in F. pratensis, but this
effect was genotype dependent. It has been observed in
more frost tolerant genotypes that low-temperature flood-
ing improved also their resistance to low-temperature
induced photoinhibition of photosynthesis (Jurczyk et al.
2013).
This study is an attempt at understanding the mecha-
nisms of freezing tolerance changes under low-temperature
flooding. The aim of the study was to verify the hypothesis
that photosynthetic apparatus response to low-temperature
flooding may contribute to changes in the freezing
tolerance.
Materials and methods
Plant material and growth conditions
The experiment was performed on the clones of 20 geno-
types of Lolium perenne (Lp) cv. Arka (Lp1-20) and 20
genotypes of Festuca pratensis (Fp) cv. Skra (Fp1-20). The
genotypes were randomly chosen from a large population
of plants, each germinated from a single seed. Seed germi-
nation and seedling growth were carried out under
controlled-environment conditions (20 °C, 12/12 h photo-
period, 200 lmol m 2 s 1 PAR provided by Agro HPS
lamps, Philips). The plants were grown in 18 cm diameter
pots, and the substrate was a mixture of loam
soil : sand : peat (1 : 1 : 1; v : v : v). Water was supplied
as required, and the plants were fertilized once a week with
a half-strength Hoagland’s solution. Relative humidity in
the growth chamber was maintained at about 60 %. When
the plants were 6 months old, plants of each genotype were
divided into five clones, two of them were used for flooding
50
and two for a control (two clones of approx. 20 tillers each
per treatment per genotype) and both groups were sub-
jected to CA (21 days at 4/2 °C, 12/12 h photoperiod,
200 lmol m 2 s 1 PAR provided by Agro HPS lamps,
Philips). The flooded plants during CA were permanently
submerged in tap water filling the pots (2 cm above the soil
level). Control plants were supplied with water as required.
When the plants were 7 months old, the experiment
was repeated after cloning the fifth clone for four clones
of approx. 20 tillers each, as described for the first
experiment.
Freezing tolerance
Freezing tolerance was determined after 21 days of CA in
the control and flooded plants as described by Kosmala
et al. (2009). Ten samples from two clones of each geno-
type (five parallel samples from each clone, about 3-cm
fragments from the middle part of the youngest, but fully
developed leaf) were collected from the flooded and control
plants grown in the growth chamber under CA conditions.
The samples were placed into 20 cm3 plastic vials on ice
(5 cm3 of frozen deionized water) to ensure ice nucleation.
Then, the samples were frozen in darkness at a cooling rate
of 0.2 °C min 1 to the testing temperatures ( 6, 10 and
14 °C), which were maintained for 1.5 h. After freezing,
the temperature was increased up to 0 °C at a rate of
3 °C h 1. The samples were thawed on a laboratory shaker,
and electrical conductance measurements were carried out
to estimate the leaf tissue damage (CC501, Elmetron,
Zabrze, Poland). The leaves were frozen in liquid nitrogen
to achieve 100 % electrolyte leakage. The percentage of
electrolyte leakage (%EL) was determined as described by
Flint et al. (1967), and tEL50 was calculated using linear
regression fitted to the central (linear) part of the relation-
ship between the freezing temperature (at least three
points) and the injury rate (%EL). The results are the
means of two experimental replicates (for each freezing
temperature four replicates representing four clones per
treatment, within each replicate five parallel samples).
Chlorophyll fluorescence studies
The measurements of chlorophyll a fluorescence were per-
formed with the Handy PEA fluorometer (Hansatech,
Kings Lynn, UK) after 21 days of CA in the control and
flooded plants. Before measurements, the LED light source
of the fluorometer was calibrated using a SQS light meter.
The measurements were taken in the growth chamber
under CA conditions within the middle section of the
youngest, but fully developed leaf. Before measurements,
the leaves were dark adapted for 30 min in a leaf clip
(Hansatech, with a 4-mm diameter hole). Polyphasic
© 2014 Blackwell Verlag GmbH, 201 (2015) 49–56
 Flooding Changes Freezing Tolerance through Photosynthesis

chlorophyll a fluorescence transients (Strasser et al. 1995)

were determined with light pulse intensity of (a)
3000 lmol m 2 s 1, pulse duration of 1 s and fixed gain
(90.8). Chlorophyll fluorescence parameters were calcu-
lated using HANDY PEA v.1.3 software (Hansatech, Kings
Lynn, UK) based on the theory of energy flow in photosys-
tem II, and using the equations of the JIP test (Strasser and
Tsimilli-Michael 2001, Strasser et al. 2005, Rapacz 2007).
These included three groups of parameters: (i) specific
energy fluxes (per QA-reducing photosystem II reaction
centre, RC), ET0/RC (electron transport flux per RC,
at t = 0) and DI0/RC (dissipated energy flux per RC, at
t = 0), (ii) yields or flux ratios, ET0/TR0 (probability at
t = 0 that a trapped exciton moves an electron into the (b)
electron transport chain beyond QA ), ET0/ABS (quantum
yield of electron transport, at t = 0), TR0/ABS (Fv/Fm,
quantum yield of energy trapping in photosystem II) and
(iii) the PSII performance index (PI) that takes into
account three main functional steps in PSII (light energy
absorption, trapping of excitation energy and a conversion
of energy to electron transport). The results are the means
of two experimental replicates (four replicates representing
four clones per treatment, within each replicate five parallel
measurements).

Statistical analysis
The statistical analysis was performed based on the results
of the measurements taken for the independent clones of
the genotypes in the first or second repetition of the experi-
ment. The tEL50 values were calculated from a linear regres-
sion fitted to the central (linear) part of the sigmoid
relationship between the freezing temperatures and electro-
lyte leakage, using at least three freezing temperatures with
coefficient intervals for P = 0.05.
The effect of flooding on the chlorophyll fluorescence
parameters for a particular genotype was tested using one-
way ANOVA (with treatment as a factor) at P = 0.05. The
general effects of flooding on the chlorophyll fluorescence
parameters were tested using two-way ANOVA (genotype
and treatment as factors) at P = 0.05. In addition, the stan-
dard error of chlorophyll fluorescence parameters was
calculated (STATISTICA 10.0, Statsoft, Tulsa, OK, USA).
Results
Fig. 1 The effect of low-temperature flooding on the frost tolerance of
(a) Lolium perenne and (b) Festuca pratensis. Eight clones of each geno-
type were divided into two groups (flooded and control), and both
groups were subjected to CA (21 days at 4/2 °C, 12/12 h photoperiod,
200 lmol m 2 s 1 PAR provided by Agro HPS lamps, Philips). tEL50
means the temperature at which 50 % of the total electrolytes were
released from the leaf tissues. The values of tEL50 and the coefficient
intervals for P = 0.05 were calculated on the basis of freezing tests
made in four replicates per treatment, within each replicate five parallel
samples, at three different temperatures ( 6, 10, 14 °C). Asterisked
values are different for the flooded and control plants (P = 0.05).
types, tEL50 was, respectively, by 3.3, 3.9 and 4.6 °C higher
than in the control plants. In Lp1 genotype, the freezing
tolerance was enhanced by flooding (tEL50 was by 2.8 °C
lower in the flooded plants) (Fig. 1a). In two Fp genotypes
(Fp4, Fp12), the freezing tolerance was decreased by flood-
ing. For the flooded Fp4 and Fp12 genotype, tEL50 was,
respectively, by 2.6 and 2.4 °C higher than in the control
plants (Fig. 1b).

The effect of flooding on freezing tolerance
After 3 weeks of CA, most of the examined genotypes (Lp
and Fp) showed no differences in the freezing tolerance in
flooded vs. control plants (Fig. 1a, b). In three Lp geno-
types (Lp5, Lp12, Lp18), the freezing tolerance was reduced
by flooding. For the flooded Lp5, Lp12 and Lp18 geno-
The effect of flooding on photosynthetic apparatus
acclimation to cold
Opposite effects of flooding on chlorophyll fluorescence
parameters were observed in two analysed species (Lp and
Fp). In the case of low-temperature flooded Lp genotypes,
a general improvement in PSII PI, energy flux ratios (ET0/

© 2014 Blackwell Verlag GmbH, 201 (2015) 49–56 51

Jurczyk et al.
(a)
(b)
Fig. 2 Changes in performance index (PI) parameter after low-tempera-
ture flooding in (a) Lolium perenne and (b) Festuca pratensis. Eight
clones of each genotype were divided into two groups (flooded and
control), and both groups were subjected to CA (21 days at 4/2 °C, 12/
12 h photoperiod, 200 lmol m 2 s 1 PAR provided by Agro HPS
lamps, Philips). PI, the performance index of photosystem II. The data
represent the means from four replicates per treatment, within each
replicate five parallel samples  S.E. Asterisked values are different for
the flooded and control plants (P = 0.05).
ABS, ET0/TR0), quantum yield of energy trapping in pho-
tosystem II (TR0/ABS) and electron transport activity per
RC (ET0/RC) in relation to the control was observed
(Figs 2a, 3a, 4a, 5a and 6a). In Fp genotypes, a general
decline in these parameters was detected (P = 0.05), as
compared to the control (Figs 2b, 3b, 4b, 5b and 6b).
Energy dissipation per RC (DI0/RC) decreased in Lp and
increased in Fp genotypes under flooding (Fig. 7a, b). The
above-mentioned general effects were also observed for the
genotypes of Fp where flooding reduced the frost tolerance
(Fp4 and Fp12) (P = 0.05). Lp genotypes, in which the
frost tolerance was significantly reduced by flooding,
showed the same general tendency as those mentioned
above, with significant increase in PI, ET0/ABS and ET0/
TR0 parameters for Lp5, Lp12 and Lp18 genotypes, an
increase in ET0/RC parameter for Lp5 and Lp18 genotypes,
52
(a)
(b)
Fig. 3 Changes in ET0/ABS parameter after low-temperature flooding
in (a) Lolium perenne and (b) Festuca pratensis. Eight clones of each
genotype were divided into two groups (flooded and control), and both
groups were subjected to CA (21 days at 4/2 °C, 12/12 h photoperiod,
200 lmol m 2 s 1 PAR provided by Agro HPS lamps, Philips). ET0/ABS,
quantum yield of electron transport at t = 0. The data represent the
means from four replicates per treatment, within each replicate five
parallel samples  S.E. Asterisked values are different for the flooded
and control plants (P = 0.05).
a reduction in DI0/RC for Lp12 and Lp18 genotypes and a
gain in TR0/ABS for Lp12 genotype (P = 0.05). Lp1 geno-
type, in which the freezing tolerance was positively affected
by flooding, showed entirely different response in the fluo-
rescence parameters when compared to other Lp genotypes.
A decrease in PI, ET0/ABS and ET0/TR0, together with an
increase in DI0/RC, was detected in the flooded Lp1 plants
in relation to the control (Figs 7a, 2a, 3a and 4a).
Discussion
Only a few studies have analysed the impact of low-temper-
ature flooding on the freezing tolerance (Rakitina 1977,
Andrews and Pomeroy 1981, Jurczyk et al. 2013). These
works yielded contrary results, and the effect of short-term
flooding on the freezing tolerance still seems to be
unpredictable.
© 2014 Blackwell Verlag GmbH, 201 (2015) 49–56

(a)
(b)
Fig. 4 Changes in ET0/TR0 parameter after low-temperature flooding in
(a) Lolium perenne and (b) Festuca pratensis. Eight clones of each geno-
type were divided into two groups (flooded and control), and both
groups were subjected to CA (21 days at 4/2 °C, 12/12 h photoperiod,
200 lmol m 2 s 1 PAR provided by Agro HPS lamps, Philips). ET0/TR0,
probability at t = 0 that a trapped exciton moves an electron into the
electron transport chain beyond QA . The data represent the means
from four replicates per treatment, within each replicate five parallel
samples  S.E. Asterisked values are different for the flooded and
control plants (P = 0.05).
The results of this study were obtained in relatively large
groups of Lp and Fp genotypes and have the potential to
reveal the species and genotype dependent reactions. Most
of the examined genotypes showed no differences in the
freezing tolerance after 3 weeks of CA in the flooded vs.
control conditions. In the case of two Fp genotypes (Fp4,
Fp12) and three Lp genotypes (Lp5, Lp12, Lp18), lowered
freezing tolerance was observed after 3 weeks of CA in the
flooded plants relative to the control ones. On the other
hand, Lp1 was the only genotype in which improved freez-
ing tolerance was observed after CA in the flooded vs. the
control plants.
Contrary to the results obtained in this study for Fp,
Jurczyk et al. (2013) showed a transient or stable
improvement in the freezing tolerance during 3 weeks of
© 2014 Blackwell Verlag GmbH, 201 (2015) 49–56
Flooding Changes Freezing Tolerance through Photosynthesis
(a)
(b)
Fig. 5 Changes in TR0/ABS parameter after low-temperature flooding
in (a) Lolium perenne and (b) Festuca pratensis. Eight clones of each
genotype were divided into two groups (flooded and control), and both
groups were subjected to CA (21 days at 4/2 °C, 12/12 h photoperiod,
200 lmol m 2 s 1 PAR provided by Agro HPS lamps, Philips). TR0/ABS,
quantum yield of energy trapping in photosystem II. The data represent
the means from four replicates per treatment, within each replicate five
parallel samples  S.E. Asterisked values are different for the flooded
and control plants (P = 0.05).
low-temperature flooding in three of four Fp genotypes. At
the end of the CA period, higher freezing tolerance was per-
ceived in only one genotype. In that experiment (Jurczyk
et al. 2013), the CA was preceded by a period of high-tem-
perature flooding (at 15 °C). It can be speculated that
high-temperature flooding, which is in fact more detrimen-
tal to plant metabolism than flooding at low temperatures
(Beard and Martin 1970), induces some mechanisms that
allow plants to survive harsh conditions. These mechanisms
may involve altered sugar metabolism (Jurczyk et al. 2013)
and may finally contribute to a better freezing tolerance.
It has been demonstrated here that under low-tempera-
ture flooding, the capacity for photosynthesis in most of Lp
genotypes was upregulated and accompanied by a drop in
the dissipation of the excess energy. These results may
indicate that a growing demand for photoassimilates,
53
Jurczyk et al.
(a)
(b)
Fig. 6 Changes in ET0/RC parameter after low-temperature flooding in
(a) Lolium perenne and (b) Festuca pratensis. Eight clones of each geno-
type were divided into two groups (flooded and control), and both
groups were subjected to CA (21 days at 4/2 °C, 12/12 h photoperiod,
200 lmol m 2 s 1 PAR provided by Agro HPS lamps, Philips). ET0/RC,
electron transport flux per RC, at t = 0. The data represent the means
from four replicates per treatment, within each replicate five parallel
samples  S.E. Asterisked values are different for the flooded and
control plants (P = 0.05).
triggered by oxygen deficiency, limited the sucrose-
depended inhibition of the photosynthetic activity.
Reduced sugar contents may have also contributed to lower
freezing tolerance in these plants.
Lp1 genotype showed a different reaction of the photo-
synthetic apparatus to low-temperature flooding than the
other Lp genotypes. In this genotype, a downregulation of
the photosynthetic capacity and increased energy dissipa-
tion were detected. This may suggest that Lp1 plants
displayed the non-photochemical mechanisms of photo-
synthetic acclimation to cold under flooding, because of
their lower demand for photoassimilates connected with
higher level of flooding (oxygen deficiency) tolerance. The
non-photochemical mechanisms of photosynthetic accli-
mation to cold were shown to be the most efficient in pro-
tecting the photosynthetic apparatus under excess light
54
(a)
(b)
Fig. 7 Changes in DI0/RC parameter after low-temperature flooding in
(a) Lolium perenne and (b) Festuca pratensis. Eight clones of each geno-
type were divided into two groups (flooded and control), and both
groups were subjected to CA (21 days at 4/2 °C, 12/12 h photoperiod,
200 lmol m 2 s 1 PAR provided by Agro HPS lamps, Philips). DI0/RC,
dissipated energy flux per RC, at t = 0. The data represent the means
from four replicates per treatment, within each replicate five parallel
samples  S.E. Asterisked values are different for the flooded and
control plants (P = 0.05).
energy (M€ uller et al. 2001). Furthermore, a relationship
between intensified non-photochemical quenching during
cold acclimation and freezing tolerance has been proved by
Humphreys et al. (2007). In their study, increased non-
photochemical quenching was detected mostly in highly
freezing tolerant genotypes of Lolium–Festuca hybrids.
Therefore, the activation of the non-photochemical mecha-
nisms under low-temperature flooding may contribute to a
better freezing tolerance observed for Lp1 and to enhanced
cold acclimation efficiency.
It has been demonstrated here that Fp genotypes under
low-temperature flooding exhibited reduced photochemi-
cal activity that was probably connected with photoinhibi-
tion of the photosystem II. Neither the non-photochemical
nor photochemical mechanisms of the photosynthetic
acclimation to cold were activated. As a consequence,
© 2014 Blackwell Verlag GmbH, 201 (2015) 49–56

diminished freezing tolerance of some Fp genotypes was
observed. Decreased photochemical activity and pho-
toinhibition found in the flooded Fp genotypes were prob-
ably triggered by metabolic disorders related to sugar
metabolism.
Humphreys et al. (2007) indicated that non-photochem-
ical quenching mechanisms for expulsion of excess energy
during CA were found in F. pratensis, but not in Lolium
multiflorum. Our results suggest that when CA process is
modified by flooding, the non-photochemical quenching
mechanisms may be activated in Lp genotypes. Moreover,
the activation of the non-photochemical quenching in Fp
may be impeded.
It may be concluded that the genotypes in which the
non-photochemical mechanisms of photosynthetic adjust-
ment to cold are activated under low-temperature flooding
may show improved freezing tolerance. On the other hand,
diminished freezing tolerance, observed under low-temper-
ature flooding, may have a different background. In Fp
genotypes, this was probably connected with reduced pho-
tochemical activity and photoinhibition of photosynthesis.
Further experiments are needed to explain freezing
tolerance drop in Lp genotypes under low-temperature
flooding.
The obtained results provide an opportunity for the
development of fast and relatively inexpensive screening
methods for determining the freezing tolerance under low-
temperature flooding. Such protocols could be used in
large population of plants to deliver genotypes resistant to
stress and could be adapted for future breeding under cli-
matic changes. They can result in fast development of new
cultivars well adapted to altered environmental conditions.
Acknowledgements
This study was supported by Polish Ministry of Science and
Higher Education, targeted subsidy for the development of
young scientists.
References
Adams, W. W., B. Demming-Adams, T. N. Rosenstiel, A. K.
Brightwell, and V. Ebbert, 2002: Photosynthesis and photo-
protection in overwintering plants. Plant Biol. 4, 545–557.
Adams, W. W., C. R. Zarter, V. Ebbert, and B. Demmig-Adams,
2004: Photoprotective strategies of overwintering evergreens.
Bioscience 54, 41–49.
Andrews, C. J., and M. K. Pomeroy, 1981: The effect of flooding
pretreatment on cold hardiness and survival of winter cereals
in ice encasement. Can. J. Plant Sci. 61, 507–513.
Beard, J. B., and D. P. Martin, 1970: Influence of water tempera-
ture on submersion tolerance of four grasses. Agron. J. 62,
257–259.
© 2014 Blackwell Verlag GmbH, 201 (2015) 49–56
Flooding Changes Freezing Tolerance through Photosynthesis
Bradford, K. J., 1983: Effect of soil flooding on leaf gas exchange
of plants. Plant Physiol. 73, 475–479.
Dat, J. F., N. Capelli, H. Folzer, P. Bourgeade, and P.-M. Badot,
2004: Sensing and signalling during plant flooding. Plant
Physiol. Biochem. 42, 273–282.
Flint, H. J., B. R. Boyce, and D. J. Brattie, 1967: Index of injury,
a useful expression of freezing injuries to plant tissues as
determined by the electric method. Can. J. Plant Sci. 47,
229–239.
Humphreys, M. W., D. Gaz sior, A. Le
sniewska-Bocianowska, Z.
Zwierzykowski, and M. Rapacz, 2007: Androgenesis as a
means of dissecting complex genetic and physiological con-
trols: selecting useful gene combinations for breeding freezing
tolerant grasses. Euphytica 158, 337–345.
H€ €
uner, N. P. A., G. Oquist, V. Hurry, M. Krol, S. Falk, and M.
Griffith, 1993: Photosynthesis, photoinhibition and low tem-
perature acclimation in cold tolerant plants. Photosynth. Res.
37, 19–39.
H€uner, N. P. A., D. P. Maxwell, G. R. Gray, L. V. Savitch, M.
Krol, A. G. Ivanov, , and S. Falk, 1996: Sensing environmental
temperature change through imbalances between energy sup-
ply and energy consumption: redox state of photosystem II.
Physiol. Plant. 98, 358–364.
Jurczyk, B., T. Krez pski, A. Kosmala, and M. Rapacz, 2013: Dif-
ferent mechanisms trigger an increase in freezing tolerance in
Festuca pratensis exposed to flooding stress. Environ. Exp.
Bot. 93, 45–54.
Kosmala, A., A. Bocian, M. Rapacz, B. Jurczyk, and Z. Zwierzy-
kowski, 2009: Identification of leaf proteins differentially
accumulated during cold acclimation between Festuca praten-
sis plants with distinct levels of frost tolerance. J. Exp. Bot. 60,
3595–3609.
M€ uller, P., X.-P. Li, and K. K. Niyogi, 2001: Non-photochemical
quenching. A response to excess light energy. Plant Physiol.
125, 1558–1566.
€
Oquist, G., and N. P. A. H€ uner, 2003: Photosynthesis of over-
wintering ever-green plants. Ann. Rev. Plant Biol. 54,
329–355.
Osborn, T. J., and M. Hulme, 2002: Evidence for trends in heavy
rainfall events over the UK. Philos. Trans. A Math. Phys. Eng.
Sci. 360, 1313–1325.
Rakitina, Z. G., 1977: Effect of a surrounding ice crust on winter
wheat plants as a function of their flooding before freezing.
Sov. Plant Physiol. 254, 317–324.
Rapacz, M., 2007: Chlorophyll a fluorescence transient during
freezing and recovery in winter wheat. Photosynthetica 45,
409–418.
Rapacz, M., D. Gaz sior, Z. Zwierzykowski, A. Le
sniewska-
Bocianowska, M. W. Humphreys, and A. P. Gay, 2004:
Changes in cold tolerance and the mechanisms of
acclimation of photosystem II to cold hardening generated
by anther culture of Festuca pratensis x Lolium multiflorum
cultivars. New Phytol. 162, 105–114.
Rapacz, M., D. Gaz sior, J. Ko
scielniak, A. Kosmala, Z. Zwierzy-
kowski, and M. W. Humphreys, 2007: The role of the
55
Jurczyk et al.
photosynthetic apparatus in cold acclimation of Lolium multi-
florum: characteristics of novel genotypes low-sensitive to PSII
over-reduction. Acta Physiol. Plant. 29, 309–316.
Strasser, R. J., A. Strivastava and Govindjee, 1995: Polyphasic
chlorophyll a fluorescence transient in plants and cyanobacte-
ria. Photochem. Photobiol. 61, 32–42.
Strasser, R. J., and M. Tsimilli-Michael, 2001: Stress in plants,
from daily rhythm to global changes, detected and quantified
by the JIP-Test. Chimie Nouvelle (SRC) 75, 3321–3326.
56
Strasser, R. J., M. Tsimilli-Michael, and A. Srivastava, 2005:
Analysis of the chlorophyll a fluorescence transient. In: G. C.
Papageorgiou and Govindjee, eds. Advances in Photosynthesis
and Respiration. Series Volume Title, Chlorophyll a Fluores-
cence: A Signature of Photosynthesis. pp. 321–362. Kluver
Acad. Publ., Dordrecht, the Netherlands.
Wample, R. L., and R. K. Thornton, 1984: Differences in the
responses of sunflower (Helanthus annuus) subjects to flood-
ing and drought stress. Physiol. Plant. 61, 611–616.
© 2014 Blackwell Verlag GmbH, 201 (2015) 49–56