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TMP A535
TMP A535
9 Springer-Verlag 1996
1,4
100_ =
~ 0.9
<1 80 -- E 1.2
o c
v
=0.8 80 o
~
o
5 A "
40
20
o
~
~ D.8
A (9
0.7 \.
Z o.e
, i i i
0 uJ
800
B ~ 0.4
8.0 c
v 700 o 0 Confrol K=3OnM r=0.96
rj
= 4.o
~ 400
300
o
Free herbicide (1/p~M)
i , i , i i , r
100 80 60 40 20
Fig. 2. Double reciprocal plots for the binding of the herbicide
Wc, % of control vcllue ionyxil to thylakoids from leaves of control (control) and water-
stressed (WS) pea plants. The Wc of stressed plants was 37%. K,
dissociation constant for the herbicide binding; r, correlation coeffic-
_~ 1oo ~C ient. Experiments were performed three times, each with two repli-
cates. The plots represent a typical result
>o 8o
~ 6o
9- ~ . Con Control Drought
20 e~ 559 nm 5 5 9 nrn
e~ e W S
o A
0 20 40 60 80
- Pt
IlluminotTon time (min)
r
during drought stress. A The Fv/Fm ratio of leaf samples (A) and
electron transfer activity (&), measured as DPC to DCPIP photo-
reduction at 600 nm, of isolated thylakoids from plants at different 0 |1
Table 1. Fv/Fm ratio, electron transfer activity (ET; Control value was 455 txmol'm- 2"s- x of reduced DCPIP), and cytochrome b559content
of leaves from control, water-stressed (WS; Wc 40% and 23%), and water-stressed/strong illumination (WS + HL; Wc 40%) pea plants before
and after re-watering (recovery). Recovery time was 2 d from rewetting at initial Wc 40% and 6 d from rewetting at initial Wc 23%.
Experiments were performed three times each with two replicates. Data are means (_ SE) for Fv/Fm from three replicates
Wc Fv/Fm ET (% of control) Chl/Cyt b559
before after before after before after before after
recovery recovery recovery recovery
Control 100% 100% 0.870 + 0.005 0.870 + 0.005 100 100 314 314
WS1 40% 80% 0.810 + 0.006 0.800 + 0.013 35 65 488 335
WS2 23% 41% 0.798 + 0.018 0.762 _ 0.033 22 43 500 351
WS1 + HL 40% 35% 0.654 + 0.038 0.633 +_0.041 0.5 0.5 480 500
et al. 1989; Allen 1992; Elich et al. 1993) the recovery in 1993, 1994; H o r t o n and Ruban 1992; Giardi 1993a, b;
the first few days was found to be accelerated (data not H u r r y et al. 1993; for a recent review on phosphorylation,
shown). see Allen 1992). In line with this idea, it has been suggested
that D1 turnover has a physiological significance leading
to downregulation of PSII activity under high light stress
Discussion (Mattoo and Edelman 1985; Oquist et al. 1992; Critchley
and Russell 1994; O s m o n d 1994; Russell et al. 1995) be-
In the present work we demonstrated that drought (the sides constituting a damage-repair cycle (for review, see
decrease of Wc by 40%) affects photosynthesis at the level Barber and Anderson 1992). In-vitro phosphorylation of
of PSII, causing a considerable depletion of the PSII core, PSII proteins has been shown to protect against irrevers-
enhanced CP43 and D1 protein degradation, and en- ible light inactivation of PSII function (photoinhibition,
hanced phosphorylation of PSII core proteins, most prob- H o r t o n and Lee 1985); however, the contribution of each
ably by activating the kinase activity. This reduction in PSII core polypeptide seems to be different (Giardi et al.
the PSII core complex is accompanied by a reorganiza- 1994).
tion of the remaining PSII to a functional form since the The lower steady-state level of D1 and CP43 caused by
F v / F m ratio was high as well as the electron transfer from water stress parallels their enhanced phosphorylation,
D P C to D C P I P , when considered per unit of protein. suggesting that phosphorylation of these proteins, but not
Thus, the "surviving" PSII core seems functionally reor- of D2 and L H C I I , m a y be a committed step for their
ganized. These observations are in accordance with pre- degradation. The role of in-vivo phosphorylation in the
vious results (Bj6rkmann and Powles 1984; Ogreen and dynamics or function of any of the PSII core proteins is
Oquist 1984; Masojidek et al. 1991) showing that drought not well defined. D1 phosphorylation has been linked to
halved the active PSII reaction centres but are at variance D1 degradation (Callahan et al. 1990; Elich et al. 1992),
with the conclusion drawn by Meyer and Kouchkovsky protection of D1 against photoinhibition (Aro et al. 1993)
(1993) that the inactivation is not due to a physical de- and to chromatic adaptation of photosystems (Elich et al.
struction of the PSII core. Our results further show that 1993).
thylakoids from water-stressed plants are highly suscep- The results presented here indicate that altering the
tible to high irradiance, as was also observed previously phosphorylation state of PSII core proteins could have
(Masojidek et al. 1991; Masojidek and Hall 1992). It seems direct consequences on D1 turnover and on the half-lives
possible that this synergism of light and water stress is the of the PSII core proteins as well. These indications are
reason for the reduced content of PSII core. Thus, the consistent with other reports. For instance: (i) In-vitro
drought-stress syndrome is a combination of two compo- phosphorylation of the peripheral antenna proteins and
nents. One, a water-stress effect, enhanced by illumina- psbH protein protects electron transfer from photoinhibi-
tion, that leads to a disassembly of a part of PSII core. tion (Horton and Lee 1985; Giardi et al. 1993, 1994). (ii) A
Second, a reorganization process that rebuilds and main- heterogeneity of PSII core phosphorylation has been
tains the remaining PSII functional to counteract the shown. Four differently phosphorylated PSII core popu-
depletion of PSII core. The importance of these two lations, which are interconvertible, are present in grana
drought consequences depends on specific growth membranes (Giardi et al. 1991, 1994). Thylakoids with
conditions and the intensity of light, offering an explana- highly phosphorylated forms of PSII core result in PSII
tion for the conflicting results seen in water-stress experi- disassembly within a few minutes of illumination (Giardi
ments carried out in a growth chamber versus those car- 1993a). (iii) In-vivo phosphorylation of D1 is an integral
ried out under field conditions (Kaiser 1987; Cornic et al. part of light-mediated D1 metabolism (Elich et al. 1992).
1992; Jefferies 1994). Recently, it has been proposed that D1 turnover is regu-
Phosphorylation of PSII polypeptides as well as D1 lated by a light-dependent kinase activity (Bracht and
turnover have been proposed as two protective mecha- Trebst 1994).
nisms for the structural and functional integrity of PSII Greatly enhanced synthesis of D1 in water-stressed
(Mattoo et al. 1989; G o d d e et al. 1991; Giardi et al. 1991, leaves and yet a lower D1 content suggest that a decreased
124 M.T. Giardi et al.: Long-term drought stress
a m o u n t of D1 is due to its e n h a n c e d degradation. The Bracht E, Trebst A (1994) Hypothesis of the control of D1 turnover
higher rate of synthesis m a y reflect preferential t r a n s l a t i o n by nuclear coded proteins in Chlamydomonasreinhardii. Z Natur-
of psbA message in response to greater depletion of the forsch 49c: 439-446
p r o t e i n as seen in senescing leaves (Droillard et al. 1992). Callahan FE , Wergin WP, Nelson N, Edelman M, Mattoo, AK
(1989) Distribution of thylakoid proteins between stromal and
This response m a y be a n a t t e m p t of water-stressed plants grana lamellae in Spirodela. Plant Physiol 91:629 635
to avoid complete depletion of the core. G i v e n the de- Callahan FE, Ghirardi ML, Sopory SK, Mehta AM, Edelman M,
pletion of the PSII core u n d e r water-stress conditions, it Mattoo AK (1990) A novel metabolic form of the 32 kDa-D1
seems reasonable to suggest that u n d e r a d d i t i o n a l stress protein in the grana-localized reaction center of photosystem II.
due to p h o t o i n h i b i t o r y light, D1 d e g r a d a t i o n m a y be J Biol Chem 265:15357 15360
altered a n d thus, synthesis, being d e p e n d e n t o n the rate of Chaves MM (1991) Effects of water deficits on carbon assimilation.
J Exp Bot 42:1 16
d e g r a d a t i o n , is reduced, as has also been shown by G e i k e n
Comic G, Ghashghaie J, Genty B, Briantais J-M (1992) Leaf photo-
et al. (1992), S u n d b y et al. (1993), a n d Russell et al. (1995). synthesis is resistant to a mild drought stress. Photosynthetica
Similar results have been recently observed u n d e r min- 27:295 309
eral stress, where also loss of active PSII centres (observed Critchley C (1988) The molecular mechanism of photoinhibition,
by fast fluorescence decay), a n increased sensitivity to facts and fiction. Aust J Plant Physiol 15:27-41
i l l u m i n a t i o n , a n d e n h a n c e d D1 protein t u r n o v e r were Critchley C, Russell AW (1994) Photoinhibition of photosynthesis in
observed (Godde a n d Hefer 1994). O u r results o n the vivo: the role of protein turnover in photosystem II. Physiol
Plant 92:188 196
effect of water stress o n the d y n a m i c s of D1 m e t a b o l i s m
Draber W, Tietjen K, Kluth JF, Trebst A (1991) Herbicides in
a n d P S I I core p h o s p h o r y l a t i o n indicate that D1 metabol- photosynthesis research. Angew Chem Int Ed Engl 30:1621-1633
ism is very i m p o r t a n t for stress a d a p t a t i o n of plants. This Droillard MJ, Bate NJ, Rothstein S, Thompson, JE (1992) Active
suggestion is s u p p o r t e d by the o b s e r v a t i o n that P S I I core translation of the D1 protein of photosystem II in senescing
depletion is reversed in a relatively short time after re- leaves. Plant Physiol 99:589 594
watering, more so when b o t h D1 d e g r a d a t i o n a n d phos- Eckert H-J, Liu B, Geiken B, Eichler H-J, Renger G (1992) Photo-
p h o r y l a t i o n are decreased by the presence of the herbicide inactivation of electron transfer through PSII as studied by time
atrazine. Thus, we suggest that the biochemical response resolved reaction centre proteins as a function of light. Photo-
synthetica 27:355 368
at the level of D1 t u r n o v e r could act as a general a d a p t a - Elich TD, Edelman M, Mattoo AK (1992) Identification, character-
tion signal for the p l a n t in response to e n v i r o n m e n t a l ization and resolution of the in vivo phosphorylated form of the
stress. D1 photosystem II reaction center protein. J Biol Chem 267:
3523-3529
This work was supported by the Italian National Council of Re- Elich TD, Edelman M, Mattoo AK (1993) Dephosphorylation of
search special grant RAISA, subproject 2 (paper No. 2179) on water photosystem II core proteins is light-regulated in vivo. EMBO
stress B. Geiken was supported by the European program "Human J 12:4857-62
Capital and Mobility". We thank Dr. Roberto Barbato (Department Geiken B, Critchley C, Renger G (1992) The turnover of photosys-
of Biology, University of Padua, Italy) for generous gifts of various tem II reaction centre proteins as a function of light. In: Murata
PSII antibodies. N (ed) Research in photosynthesis, vol 4. Kluwer Academic
Publishers, Dordrecht, pp 643-646
Genty B, Briantais J-M, Vieira da Silva JB (1987) Effects of drought
on primary photosynthetic processes of cotton leaves. Plant
References Physiol 83:360-364
Giardi MT (1993a) Phosphorylation and disassembly of photosys-
Allen JF (1992) Protein phosphorylation in regulation of photosyn- tern II core as an early stage of photoinhibition. Planta 190:
thesis. Biochim Biophys Acta 1098:275 335 107 113
Anderson JM, Aro EM (1994) Grana stacking and protection of Giardi MT (1993b) Significance of photosystem II core phos-
photosystem II in thylakoid membranes of higher plants under phorylation heterogeneity for the herbicide binding domain.
sustained high irradiance. Photosynth Res 41:315-326 Z Naturforsch 48c: 241 245
Aro EM, Virgin I, Andersson B (1993) Photoinhibition of photo- Giardi MT, Rigoni F, Barbato R, Giacometti GM (1991) Relation-
system II. Inactivation, protein damage and turnover. Biochim ships between heterogeneity of PSII in grana particles in vitro
Biophys Acta 1143:113-134 and phosphorylation. Biochem Biophys Res Comm 176:
Baker NR (1991) Possible role of photosystem II in environmental 1298-1305
perturbations of photosynthesis. Physiol Plant 81:563-570 Giardi MT, Rigoni F, Barbato R (1992) PSII core phosphorylation
Barbato R, Friso G, Giardi MT, Rigoni F, Giacometti GM (1991) heterogeneity: differential herbicide binding and electron transfer
Breakdown of the photosystem II reaction center D1 protein regulation in PSII particles from spinach. Plant Physiol 100:
under photoinhibitory conditions: identification and locali- 1948 1954
zation of the C-terminal degradation product. Biochemistry 30: Giardi MT, Komenda J, Masojidek J (1994) Involvement of PSII
10020-10026 phosphorylation in the sensitivity to strong light. Physiol Plant
Barber J, Andersson B (1992) Too much of a good thing: light can be 92:181-187
bad for photosynthesis. TIBS 17:61 66 Godde D, Hefer M (1994) Photoinhibition and light dependent
Beadle CL, Ludlow MM, Honeysett JL (1993) Water relations. In: turnover of D 1 reaction-centre polypeptide of photosystem II are
Hall, DO et al. (eds) Photosynthesis and production in a chang- enhanced by mineral-stress conditions. Planta 193, 290 299
ing environment: a field and laboratory manual. Chapman Godde D, Schmitz H, Weiner M (1991) Turnover of the D1 reaction
& Hall, London, pp t13 128 center polypeptide from photosystem lI in intact spruce needles
Bj~Srkmann O, Powles SB (1984) Inhibition of photosynthetic reac- and spinach leaves. Z Naturforsch 46c: 245 251
tions under water stress: interaction with light level. Planta 161: Havaux M (1992) Stress tolerance of photosystem II in vivo. Plant
490-504 Physiol 100:424 432
Boyer JS, Bowen BL (1970) Inhibition of oxygen evolution in Horton P, Lee P (1985) Phosphorylation of chloroplast membrane
chloroplast isolated from leaves with low water potentials. Plant proteins partially protects against photoinhibition. Planta 165:
Physiol 45:612-615 37 42
M.T. Giardi et al.: Long-term drought stress 125
Horton P, Ruban AV (1992) Regulation of PSII. Photosynth Res 34: thylakoids of drought resistant and sensitive lupin plants. Photo-
375-385 synth Res 37:49-60
Hurry VM, Gardestr/Sm P, Oquist G (1993) Reduced sensitivity to Ogren E, Oquist G (1984) Photoinhibition of photosynthesis in
photoinhibition following frost-hardening of winter rye is due to Lemna gibba as induced by the interaction between light and
increased phosphate availability. Planta 190:484-490 temperature. III Chlorophyll fluorescence at 77 K. Physiol Plant
Jefferies RA (1994) Drought and chlorophyll fluorescence in field- 62:193 200
grown potato (Solanum tuberosus). Physiol Plant 90:93-97 Oquist G, Chow WS, Anderson M (1992) Photoinhibition of photo-
Kaiser WM (1987) Effects of water deficit on photosynthetic capa- synthesis represents a mechanism for the long-term regulation of
city. Physiol Plant 71:142 149 photosystem II. Planta 186:450 460
Lichtenthaler H K, Wellburn AR (1983) Determination of total caro- Osmond B (1994) What is photoinhibition: some insights from
tenoids and chlorophyll a and b of leaf extracts in different comparisons of sun and shade plants. In: Baker NR, Bowyer JR
solvents. Biochem Soc Trans 603:591-592 (eds) Photoinhibition: Molecular mechanisms to the field. Bios
Masojidek J, Hall DO (1992) Salinity and drought stresses are ampli- Science Publisher, Oxford, pp 1 24
fied by high irradiance in sorghum. Photosynthetica 27:159-171 Powles SB (1984) Photoinhibition of photosynthesis induced by
Masojidek J, Trivedi S, Halshaw L, Alexiou A, Hall DO (1991) The visible light. Annu Rev Plant Physiol 35:15 44
synergistic effect of drought and light stresses in sorghum and Russell AW, Critchley C, Robinson SA, Franklin LA, Seaton, GGR,
pearl millet. Plant Physiol 96:198 207 Chow WS, Anderson JM, Osmond CB (1995) Photosystem II
Mattoo AK, Edelman M (1985) Photoregulation and metabolism regulation and dynamics of the chloroplast D1 protein in Ara-
of a thylakoidal herbicide-receptor protein. In: John SB (ed) bidopsis leaves during photosynthesis and photoinhibition. Plant
Frontiers of membrane research in agriculture. Rowman and Physiol 107:943-952
Allanheld, Totowa, pp 23-24 Sundby C, Mc Caffery S, Anderson JM (1993) Turnover of the
Mattoo AK, Marder JB, Edelman M (1989) Dynamics of the photo- photosystem II D1 protein in higher plants under photoinhibi-
system II reaction center. Cell 56:241-246 tory and non-photoinhibitory irradiance. J Biol Chem 268:
Meyer S, de Kouchkovsky Y (1993) Electron transport, photosys- 25476-25482
tem-2 reaction centers and chlorophyll-protein complexes of