The role of Ca ions in changes induced by excess Cu2+ in bean plants. Growth parameters

The effect of Ca on Cu toxicity in runner bean plants (Phaseolus coccineus L. cv. Piěkny Jaś) grown hydroponically in nutrient solution was studied. The toxic effect of excess Cu on plants depends on their age and Ca content in the medium. Copper applied in excess to the plants at the early phase of leaf development strongly limits the uptake of Ca ions from the nutrient solution, particularly their translocation to leaves. Increased Ca content limits the inhibitory effect of Cu on leaf growth and decreases the content of chloroplast pigments to the level approximate to that of control. At this growth stage the effect of excess Cu is at least partially connected with limited Ca transport to leaves. At the intermediate leaf phase Cu-treated plants react slightly to changed Ca content. At the end of the primary leaf development increased Ca concentration in the medium intensifies senescence processes induced by excess Cu. The changes are partially connected with intensified water deficit. Increased Ca content in the nutrient solution limits Cu accumulation in the individual organs of Cu-treated plants. However, Cu accumulation in leaves is not decreased at a high level of Ca. Copper generally decreases Ca content in the youngest plants, whereas in the oldest ones only in the case of a low level of Ca in the nutrient solution.     

Photoprotective function of betacyanin in leaves of <Emphasis Type="Italic">Amaranthus cruentus</Emphasis> L. under water stress

The photoprotective function of leaf betacyanin in water-stressed Amaranthus cruentus plants was examined by comparing leaves of two strains which differ significantly in the amount of betacyanin. At 0, 1, and 2 days after the imposed water stress, leaves were subjected to high-light (HL) treatment to assess their photosynthetic capacity and photoinhibition susceptibility. The water stress equally reduced leaf relative water content (RWC), gas-exchange rate and chlorophyll (Chl) contents in both leaves, indicating that the severity of water stress was comparable between the strains. Consequently, the extent of photoinhibition after the HL treatment increased in both strains as water stress developed; however, it was significantly greater in acyanic leaves than in betacyanic leaves, suggesting lower photoinhibition susceptibility in the betacyanic strain. The betacyanic leaves also exhibited approximately 30% higher values for photochemical quenching coefficient (q_P) during the period of water stress despite the nonphotochemical quenching coefficient (q_N) did not differ significantly between the strains. These results may be partially explained by the increased amount of leaf betacyanin under water stress. Moreover, a decrease in Chl content in betacyanic leaves might have enhanced light screening effect of betacyanin by increasing relative abundance of betacyanin to Chl molecule. In addition, reduced Chl content increased light penetrability of leaves. As a result, the extent of photoinhibition at the deeper tissue was exacerbated and the Chl fluorescence emitted from these tissues was more readily detected, facilitating assessment of photoinhibition at deeper tissues where the effect of betacyanic light screening is considered to be most apparent. Our results demonstrated that leaf betacyanin contributes to total photoprotective capacity of A. cruentus leaves by lowering excitation pressure on photosystem II (PSII) via attenuation of potentially harmful excess incident light under water stress.     

Effects of copper on the growth,photosynthesis and nutrient concentrations of Phaseolus plants

Bean plants (Phaseolus vulgaris L. var. Zargana Kavala) were grown under conditions of increasing Cu concentrations in the growth medium (0.5-160.5 µM). Generally, the Cu concentrations between 0.5-1.5 µM were deficient, 1.5-10.5 µM were optimal, and 10.5-160.5 µM were toxic to plant growth. The Cu toxicity was associated with marked increases in plant tissue Cu concentrations. Under the Cu-deficient and optimal growth conditions, Cu was located primarily in the leaves. Under Cu toxicity, it was primarily sequestered in the roots. With increasing Cu in the growth medium, there was a positive correlation between Cu concentrations in the roots, stems and leaves, Ca in the roots, and K and Mg in the leaves. In contrast, Ca concentrations in the leaves and stems showed a negative correlation. The chlorophyll (Chl) concentration increased with increasing leaf Cu concentration, however, the Chl a/b ratio decreased. Since with an increasing leaf Cu concentration the leaf area decreased more markedly than the leaf dry mass, the net photosynthetic rate (PN) per leaf area increased and per dry mass decreased. The increase in PN per leaf area was almost entirely accounted for by the increase in Chl concentration. The initial Chl fluorescence (F0) increased with increasing leaf Cu concentration. The ratio of variable to maximum fluorescence (Fv/Fm) under Cu toxicity decreased. The half-time for the rise from F0 to Fm (t1/2) remained relatively unchanged with increasing leaf Cu concentration. Therefore the Cu-stress caused a small decrease in the efficiency of photosystem 2 photochemistry, but its primary effect was on growth.     

Effects of copper and zinc ions on photosystem II studied by EPR spectroscopy.

The effect of Zn(2+) or Cu(2+) ions on Mn-depleted photosystem II (PS II) has been investigated using EPR spectroscopy. In Zn(2+)-treated and Cu(2+)-treated PS II, chemical reduction with sodium dithionite gives rise to a signal attributed to the plastosemiquinone, Q(A)(*)(-), the usual interaction with the non-heme iron being lost. The signal was identified by Q-band EPR spectroscopy which partially resolves the typical g-anisotropy of the semiquinone anion radical. Illumination at 200 K of the unreduced samples gives rise to a single organic free radical in Cu(2+)-treated PS II, and this is assigned to a monomeric chlorophyll cation radical, Chl a(*)(+), based on its (1)H-ENDOR spectrum. The Zn(2+)-treated PS II under the same conditions gives rise to two radical signals present in equal amounts and attributed to the Chl a(*)(+) and the Q(A)(*)(-) formed by light-induced charge separation. When the Cu(2+)-treated PS II is reduced by sodium ascorbate, at >/=77 K electron donation eliminates the donor-side radical leaving the Q(A)(*)(-) EPR signal. The data are explained as follows: (1) Cu(2+) and Zn(2+) have similar effects on PS II (although higher concentrations of Zn(2+) are required) causing the displacement of the non-heme Fe(2+). (2) In both cases chlorophyll is the electron donor at 200 K. It is proposed that the lack of a light-induced Q(A)(*)(-) signal in the unreduced Cu(2+)-treated sample is due to Cu(2+) acting as an electron acceptor from Q(A)(*)(-) at low temperature, forming the Cu(+) state and leaving the electron donor radical Chl a(*)(+) detectable by EPR. (3) The Cu(2+) in PS II is chemically reducible by ascorbate prior to illumination, and the metal can therefore no longer act as an electron acceptor; thus Q(A)(*)(-) is generated by illumination in such samples. (4) With dithionite, both the Cu(2+) and the quinone are reduced resulting in the presence of Q(A)(*)(-) in the dark. The suggested high redox potential of Cu(2+) when in the Fe(2+) site in PS II is in contrast to the situation in the bacterial reaction center where it has been shown in earlier work that the Cu(2+) is unreduced by dithionite. It cannot be ruled out however that Q(A)-Cu(2+) is formed and a magnetic interaction is responsible for the lack of the Q(A)(-) signal when no exogenous reductant is present. With this alternative possibility, the effects of reductants would be explained as the loss of Cu(2+) (due to formation of Cu(+)) leading to loss of the Cu(2+) from the Fe(2+) site due to the binding equilibrium. The quite different binding and redox behavior of the metal in the iron site in PS II compared to that of the bacterial reaction center is presumably a further reflection of the differences in the coordination of the iron in the two systems.     

Optimal copper supply is required for normal plant iron deficiency responses

Iron (Fe) and copper (Cu) homeostasis are tightly linked across biology. Understanding crosstalk between Fe and Cu nutrition could lead to strategies for improved growth on soils with low or excess metals, with implications for agriculture and phytoremediation. Here, we show that Cu and Fe nutrition interact to increase or decrease Fe and/or Cu accumulation in leaves and Fe uptake processes. Leaf Cu concentration increased under low Fe supply, while high Cu lowered leaf Fe concentration. Ferric reductase activity, an indicator of Fe demand, was inhibited at insufficient or high Cu supply. Surprisingly, plants grown without Fe were more susceptible to Cu toxicity.     

Chilling sensitivity of the frost-tolerant potato Solanum commersonii

Frost tolerance has been reported in the shoots of wild, tuberiferous potato species such as Solanum commersonii when the plants are grown in either field or controlled conditions. However, these plants can survive as underground tubers and avoid unfavorable environmental conditions altogether. As such, leaf growth and photosynthesis at low temperature may not be required for survival of the plants. In order to determine the temperature sensitivity of S. commersonii shoots, we examined leaf growth, development and photosynthesis in plants raised at 20/16°C (day/night). 12/9°C and 5/2°C. S. commersonii leaves grown at 5°C exhibited a marked decrease in leaf area and in total chlorophyll (Chl) content per leaf area when compared with leaves grown at 20°C. Furthermore, leaves grown at 5°C did not exhibit the expected decrease in either water content or susceptibility to low-temperature-induced photoinhibition that normally characterizes cold acclimation in frost-tolerant plants. Measurements of CO₂-saturated O₂ evolution showed that the photosynthetic apparatus of 5°C plants was functional, even though the efficiency of photosystem II photochemistry was reduced by growth at 5°C. A decrease in the resolution of the M-peak in the slow transients for Chl a fluorescence in leaves grown at 12 and 5°C and in all leaves exposed to high light at 5°C indicated that low temperature significantly affected processes on the reducing side of Q_A, the primary quinone electron acceptor in photosystem II. Thus S. commarsonii exhibits the characteristics of a plant that is limited by chilling temperatures. Although S. commersonii can tolerate light frosts, its sensitivity to chilling temperatures may result in shoot dieback in winter in its native habitat. The plants may avoid both chilling and freezing temperatures by overwintering as underground tubers.     

Induction characteristics and response of photosynthetic quantum conversion to changes in irradiance in mulberry plants

A study was conducted, using rapid time course of chlorophyll (Chl) fluorescence parameters, and light-response curves of Chl fluorescence parameters, to determine the induction requirements and response of photosystem II (PSII) photochemistry and non-photochemical reactions after changes in irradiance in greenhouse mulberry plants. The induction of PSII photochemistry rapidly approached to steady state after leaves were treated from darkness to low irradiance (LI). When irradiance of leaves changed from darkness to high irradiance (HI), a biphasic induction was observed. A slight photoinhibition occurred in the leaves exposed to sunlight coming to the greenhouse, whereas a chronic photoinhibition occurred in the leaves fully exposed to sunlight outside the greenhouse. The chronic photoinhibition was demonstrated by sustained reduction of maximal quantum yield of PSII photochemistry (Fv/Fm). Moreover, the leaves of mulberry plants in greenhouse were sensitive to abrupt changes in irradiance and the sensitivity of leaves suffered in a short-term (1h) high light treatment was reduced, based on the changes in photosynthetic quantum conversion. These results demonstrated an inducible response of photosynthetic quantum conversion to changes in irradiance in mulberry.     

Mechanisms of photoinhibition induced by high light in Hosta grown outdoors

Aims It has long been recognized that photoinhibition of photosynthesis is induced by high light. However, our recent studies are not consistent with this traditional view. Therefore, the objective of this study is to explore the induction of photoinhibition and its mechanisms under full sunlight outdoors.
Methods Changes of leaf morphology, gas exchange, and chlorophyll a fluorescence were measured to investigate the induction and mechanisms of photoinhibition under high light in Hosta, which is a typical shade-tolerant plant.
Important findings Hosta plants grown under full sunlight (HT) and low light (LT) developed sun- and shade-type leaf morphological characteristics, respectively. Under a full sunlight, Hosta plants had lower photosynthetic rate and chlorophyll content than under the LT; whereas, there were only slight difference in the maximum quantum yield of photosystem II (F_v/F_m) between the two treatments, suggesting that Hosta plants could grow normally under full sunlight without severe photoinhibition. After transition from the low to a high light (LHT), the photosynthetic rate and maximum quantum yield of photosystem II decreased sharply, reflecting that the LHT treatment led to irreversibly inactivation of photosystem II. Additionally, the shape of chlorophyll a fluorescence transients also changed significantly; the relative fluorescence yield of the K and J steps were reduced by 24.3% and 34.2%, respectively, indicating that the acceptor side of photosystem II was damaged more severely than the donor side. Consequently, we postulate that photoinhibition in Hosta leaves is mainly induced by the sudden enhancement of light intensity outdoors. Hosta can acclimate to high irradiance through leaf development outdoors. Our finding is of great significance in understanding the acclimation of plants to high light and cultivation of shade-tolerant plants in field.     

Leaf plasticity in peanut (Arachis hypogaea L.) in response to heavy metal stress

Phenotypic plasticity in morphological, anatomical and physiological traits of peanut (Arachis hypogaea L.) leaves was tested at four different concentrations of Cd, Cu and Zn under greenhouse conditions. Among 18 characteristics tested, nine were found to be the most sensitive and demonstrate the greatest phenotypic plasticity. These were: the leaf area (LA), the leaf mass per area (LMA), chlorophyll a concentration (Chl a), chlorophyll b concentration (Chl b), total chlorophyll concentration (Chl t), the effective quantum yield of photosystem II (ΦPS II), stomatal density of upper epidermis (SDU), palisade thickness (PT), and palisade to spongy thickness ratio (P/S). The plasticity of chlorophyll concentration and fluorescence parameters may be maladaptive and reflects metal toxicity to leaves, whereas the anatomical plasticity is adaptive, indicative of a tradeoff between the physiological and anatomic plasticity. The anatomical plasticity resulted in a xerophyte feature of leaves (i.e. small leaflets, thick lamina, upper epidermis, palisade mesophyll, as well as abundant and small stomata), which enhanced the capacity to resist drought caused by heavy metals via a decrease in root growth.     

Leaf movements and photoinhibition in relation to water stress in field-grown beans

Photoinhibition in plants depends on the extent of light energy being absorbed in excess of what can be used in photochemistry and is expected to increase as environmental constraints limit CO2 assimilation. Water stress induces the closure of stomata, limiting carbon availability at the carboxylation sites in the chloroplasts and, therefore, resulting in an excessive excitation of the photosynthetic apparatus, particularly photosystem II (PSII). Mechanisms have evolved in plants in order to protect against photoinhibition, such as non-photochemical energy dissipation, chlorophyll concentration changes, chloroplast movements, increases in the capacity for scavenging the active oxygen species, and leaf movement or paraheliotropism, avoiding direct exposure to sun. In beans (Phaseolus vulgaris L.), paraheliotropism seems to be an important feature of the plant to avoid photoinhibition. The extent of the leaf movement is increased as the water potential drops, reducing light interception and maintaining a high proportion of open PSII reaction centres. Photoinhibition in water-stressed beans, measured as the capacity to recover F(v)/F(m), is not higher than in well-watered plants and leaf temperature is maintained below the ambient, despite the closure of stomata. Bean leaves restrained from moving, increase leaf temperature and reduce qP, the content of D1 protein and the capacity to recover F(v)/F(m) after dark adaptation, the extent of such changes being higher in water-stressed plants. Data are presented suggesting that even though protective under water stress, paraheliotropism, by reducing light interception, affects the capacity to maintain high CO2 assimilation rates throughout the day in well-watered plants.     

Inhibition of photosynthetic reactions under water stress: interaction with light level

When the shrub Nerium oleander L., growing under full natural daylight outdoors, was subjected to water stress, stomatal conductance declined, and so did non-stomatal components of photosynthesis, including the CO₂-saturated rate of CO₂ uptake by intact leaves and the activity of electron transport by chloroplasts isolated from stressed plants. This inactivation of photosynthetic activity was accompanied by changes in the fluorescence characteristics determined at 77 K (-196°C) for the upper leaf surface and from isolated chloroplasts. The maximum (F _M) and the variable (F _V) fluorescence yield at 692 nm were strongly quenched but there was little effect on the instantaneous (F _O) fluorescence. There was a concomitant quenching of the maximum and variable fluorescence at 734 nm. These results indicate an inactivation of the primary photochemistry associated with photosystem II. The lower, naturally shaded surfaces of the same leaves were much less affected than the upper surfaces and water-stress treatment of plants kept in deep shade had little or no effect on the fluorescence characteristics of either surface, or of chloroplasts isolated from the water-stressed leaves. The effects of subjecting N. oleander plants, growing in full daylight, to water stress are indistinguishable from those resulting when plants, grown under a lower light regime, are exposed to full daylight (photoinhibition). Both kinds of stress evidently cause an inactivation of the primary photochemistry associated with photosystem II. The results indicate that water stress predisposes the leaves to photoinhibition. Recovery from this inhibition, following restoration of favorable water relations, is very slow, indicating that photoinhibition is an important component of the damage to the photosynthetic system that takes place when plants are exposed to water stress in the field. The underlying causes of this water-stress-induced susceptibility to photoinhibition are unknown; stomatal closure or elevated leaf temperature cannot explain the increased susceptibility.Abbreviations and symbols Chl chlorophyll - PFD photon flux area density - PSI, PSII photosystem I, II - F _M, F _O, F _V maximum, instantaneous, variable fluorescence emission - leaf water potential C.I.W.-D.P.B. Publication No. 775     

The irreversible photoinhibition of the photosystem II complex in leaves of Vicia faba under strong light

Exposure of the photosynthetic machinery to strong light causes the photoinhibition of the photosystem II complex. The recovery from the photoinhibition in vivo was characterized by monitoring the ratio of variable to maximum fluorescence (F_v/F_m) in detached leaves of broad bean (Vicia faba). The changes in the ratio were explained in terms of three components, namely, two saturating exponential components with half rise-times of about 15 and 120 min, respectively, and a non-recovery component. The non-recovery component increased gradually as the exposure to strong light was prolonged. Our results suggest that this irreversible component of the photoinhibition of the photosystem II complex was caused by severe stress due to strong light under which repair of the photosystem II complex was insufficient to allow full recovery. The irreversible photoinhibition is discussed in terms of both the physiology and ecology of plants.     

Photoinhibition in tropical forest understorey species with short- and long-lived leaves

1. Shade-tolerant species that inhabit the understorey have a range of leaf lifetimes (from 1 to 8 years), which may indicate a variety of strategies for dealing with increases in light associated with tree-fall gaps. We hypothesized that species with long-lived leaves should be more tolerant of an increase in light levels than species with short-lived leaves.
2. In understorey plants of 12 shade-tolerant rain-forest species, photoinhibition, measured as a reduction in the chlorophyll fluorescence parameter F _v/ F _m when leaf discs were exposed to 1h at 1000μmol m^–2s^–1, was greater in species with short-lived leaves than species with long-lived leaves.
3. Less photoinhibition in species with long-lived leaves was not associated with higher levels of non-photochemical dissipation (NPQ) of absorbed light, but may be the result of a higher yield of photosystem II compared with short-lived leaves.
4. Thus, species with long-lived leaves are more tolerant of abrupt increases in light that occur when tree-fall gaps are formed than species with short-lived leaves.
5. Discs from leaves of all species growing in tree-fall gaps had higher levels of NPQ, yield of photosystem II and more rapid recovery from photoinhibition than leaves developed in the understorey; however, there were no differences among species with short- and long-lived leaves.     

Seasonal photosynthetic changes in the green-stemmed Mediterranean shrub <Emphasis Type="Italic">Calicotome villosa</Emphasis>: a comparison with leaves

Some photosynthetic attributes of leaves and stems were seasonally followed in the small-leaved, summer-deciduous, green-stemmed Mediterranean shrub Calicotome villosa. Both leaves and stems displayed similar photon energy-saturated photosystem 2 (PS2) efficiencies with a minimum during winter. A second minimum in stems during the leafless summer period could be ascribed to sustained photoinhibition. Yet, stems were slightly inferior in photon capture, resulting partly from lower chlorophyll (Chl) contents and partly from higher reflectance due to pubescence. As a result, photon energy-saturated linear electron transport rates were slightly higher in leaves. However, when the total leaf and stem areas were taken into account, this superiority was abolished during autumn and winter and more than overturned during spring. Given that during summer the stems were the only photosynthetic organs, the yearly photosynthetic contribution of stems was much higher. Chl contents in stems displayed a transient and considerable summer drop, accompanied by an increase in the carotenoid to Chl ratio, indicating a photo-protective adaptation to summer drought through a decrease of photo-selective capacity, typical for leaves of many Mediterranean plants.     

Photosynthesis and photoprotection in Nicotiana tabacum L. in vitro-grown plantlets

Nicotiana tabacum L. plantlets were cultured in vitro photoautotrophically (0% sucrose) and photomixotrophically (3% or 5% sucrose) at two irradiances (80 or 380 mumol m-2 s-1) with the aim of investigating the effect of these culture conditions on photosynthetic parameters and on protective systems against excess excitation energy. In plantlets grown photoautotrophically under higher irradiance photoinhibition was demonstrated. These plantlets had a decreased chlorophyll (Chl) a + b content and Chl a/b ratio, an increased content of xanthrophyll cycle pigments and a higher deepoxidation state, a decreased maximum photochemical efficiency of photosystem II (PS II) and actual photochemical efficiency of PS II, and an increased non-photochemical quenching. In the photoautotrophically grown plantlets and those photomixotrophically grown with 3% sucrose, the increase of growth irradiance from 80 to 380 mumol m-2 s-1 stimulated the activities of ascorbate-glutathione cycle enzymes with the exception of ascorbate peroxidase. Ascorbate peroxidase activity was not affected by the increase in growth irradiance but a significant decrease with increasing sucrose concentration was evident. The higher concentration of sucrose in the medium (5%) in combination with the higher irradiance inhibited photosynthesis (decrease in Chl a + b content and net photosynthetic rate) but no significant changes in activities of ascorbate-glutathione cycle enzymes were found. These results suggest that exogenous sucrose added to the medium improved high irradiance and oxidative stress resistance of the plantlets but the effect of sucrose is concentration dependent.     

Effects of aluminium on photosynthetic performance in Al-sensitive and Al-tolerant maize inbred lines

Maize plant inbred lines, one Al-sensitive (B-73) and two Al-tolerant (F-2 and L-2039), were grown hydroponically in the presence of 200 μM Al. After 13 d of growth, root and shoot lengths, photosystem 2 (PS2) activity, chlorophyll (Chl) content, 5-aminolevulinic acid (5-ALA) synthesis rate, chlorophyllase (Chlase) activity, and N, Mg, Fe, and Mn contents in leaves were determined. PS2 activity and Chl content were most severely affected by Al in B-73, but F-2 was almost unaffected. This was in accordance with Al-accumulation in the plants. The observed changes in B-73 coincided with 5-ALA synthesis inhibition, Chlase activation, and leaf deprivation of Fe and Mg. In Al-treated L-2039 plants, the leaf Mg and Mn contents were decreased. Also, an excessive Chlase activation was found in Al-treated L-2039, without a substantial Chl loss. This may indicate the activation of different enzyme pools in tolerant and sensitive genotypes under low-stress conditions.     

Chloroplast movement provides photoprotection to plants by redistributing PSII damage within leaves

Plants use light to fix carbon through the process of photosynthesis but light also causes photoinhibition, by damaging photosystem II (PSII). Plants can usually adjust their rate of PSII repair to equal the rate of damage, but under stress conditions or supersaturating light-intensities damage may exceed the rate of repair. Light-induced chloroplast movements are one of the many mechanisms plants have evolved to minimize photoinhibition. We found that chloroplast movements achieve a measure of photoprotection to PSII by altering the distribution of photoinhibition through depth in leaves. When chloroplasts are in the low-light accumulation arrangement a greater proportion of PSII damage occurs near the illuminated surface than for leaves where the chloroplasts are in the high-light avoidance arrangement. According to our findings chloroplast movements can increase the overall efficiency of leaf photosynthesis in at least two ways. The movements alter light profiles within leaves to maximize photosynthetic output and at the same time redistribute PSII damage throughout the leaf to reduce the amount of inhibition received by individual chloroplasts and prevent a decrease in photosynthetic potential.     

A critical role for the Var2 FtsH homologue of Arabidopsis thaliana in the photosystem II repair cycle in vivo.

Using a var2-2 mutant of Arabidopsis thaliana, which lacks a homologue of the zinc-metalloprotease, FtsH, we demonstrate that this protease is required for the efficient turnover of the D1 polypeptide of photosystem II and protection against photoinhibition in vivo. We show that var2-2 leaves are much more susceptible to light-induced photosystem II photoinhibition than wild-type leaves. Furthermore, the rate of photosystem II photoinhibition in untreated var2-2 leaves is equivalent to that of var2-2 and wild-type leaves, which have been treated with lincomycin, an inhibitor of the photosystem II repair cycle at the level of D1 synthesis. This is in contrast to untreated wild-type leaves, which show a much slower rate of photosystem II photoinhibition due to an efficient photosystem II repair cycle. The recovery of var2-2 leaves from photosystem II photoinhibition is also impaired relative to wild-type. Using Western blot analysis in the presence of lincomycin we show that the D1 polypeptide remains stable in leaves of the var2-2 mutant under photoinhibitory conditions that lead to D1 degradation in wild-type leaves and that the abundance of DegP2 is not affected by the var2-2 mutation. We conclude, therefore, that the Var2 FtsH homologue is required for the cleavage of the D1 polypeptide in vivo. In addition, we identify a conserved lumenal domain in Var2 that is unique to FtsH homologues from oxygenic phototrophs.     

Enhancement of oxidative stress tolerance in transgenic tobacco plants overproducing Fe-superoxide dismutase in chloroplasts.

A chimeric gene consisting of the coding sequence for chloroplastic Fe superoxide dismutase (FeSOD) from Arabidopsis thaliana, coupled to the chloroplast targeting sequence from the pea ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit, was expressed in Nicotiana tabacum cv Petit Havana SR1. Expression of the transgenic FeSOD protected both the plasmalemma and photosystem II against superoxide generated during illumination of leaf discs impregnated with methyl viologen. By contrast, overproduction of a mitochondrial MnSOD from Nicotiana plumbaginifolia in the chloroplasts of cv SR1 protected only the plasmalemma, but not photosystem II, against methyl viologen (L. Slooten, K. Capiau, W. Van Camp, M. Van Montagu, C. Sybesma, D. Inzé [1995] Plant Physiol 107: 737-750). The difference in effectiveness correlates with different membrane affinities of the transgenic FeSOD and MnSOD. Overproduction of FeSOD does not confer tolerance to H2O2, singlet oxygen, chilling-induced photoinhibition in leaf disc assays, or to salt stress at the whole plant level. In nontransgenic plants, salt stress led to a 2- to 3-fold increase in activity, on a protein basis, of FeSOD, cytosolic and chloroplastic Cu/ZnSOD, ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase. In FeSOD-overproducing plants under salt stress, the induction of cytosolic and chloroplastic Cu/ZnSOD was suppressed, whereas induction of a water-soluble chloroplastic ascorbate peroxidase isozyme was promoted.     

A nuclear mutant of Arabidopsis thaliana selected for enhanced sensitivity to light-chill stress is altered in PSII electron transport activity

Chilling in the light imposes a considerable level of stress on the photosynthetic apparatus, resulting in a decrease of photosystem II activity and the quenching of maximum and variable fluorescence. We selected in a fah - 1 mutagenized population of Arabidopsis thaliana , which permits a direct visible evaluation of the intensity of chlorophyll (Chl) fluorescence, a monogenic recessive nuclear mutant hypersensitive to photoinhibition induced by light and cold. The major phenotypic trait of the mutant is the appearance of chlorotic areas on developed leaves. Photochemical analyses indicate that the mutant is hypersensitive to photoinhibition in excess light in the cold but also at room temperature. The susceptibility to photoinhibition is a consequence of perturbations in photochemistry already present in unstressed plants. Such perturbations result in a greater fraction of the primary acceptor Q_A remaining in the reduced state even at low light fluxes. From estimates of the number of total and functional PSII units and measurements of PSII quantum yield and Q_A⁻ reoxidation kinetics, the basic lesion of the mutant seems restricted to PSII photochemistry likely affecting the rate of electron transport from Q_A⁻ to Q_B.