Effect of drought stress on chlorophyll a fluorescence and electrical admittance of shoots in Norway spruce seedlings

Effects of mild and severe soil drought on the water status of needles, chlorophyll a fluorescence, shoot electrical admittance, and concentrations of photosynthetic pigments in needles of seedlings of Picea abies (L.) Karst. were examined under controlled greenhouse conditions. Drought stress reduced shoot admittance linearly with a decrease in shoot water potential (_w) and increase in water deficit (WD) and led to a decrease in concentrations of chlorophyll a, b and carotenoids. Severe water stress (shoot _w=–2.4 MPa) had a negative effect on chlorophyll a fluorescence parameters including PSII activity (F_v/F_m), and the vitality index (R_fd). Variations in these parameters suggest an inhibition of the photosynthetic electron transport in spruce needles. Water stress led to a decrease in the mobility of electrolytes in tissues, which was reflected by decreased shoot electrical admittance. After re-watering for 21 days the WD in needles decreased and the shoot water potential increased. In the re-watered plants, the chloroplast function was restored and chlorophyll a fluorescence returned to a similar level as in the control plants. This improved hydraulic adjustment in the seedlings triggered a positive effect on ion flow in the tissues and increased shoot electrical admittance. We conclude that the shoot electrical admittance and photosynthetic electron transport in leaves are closely linked to changes in water status and their decrease is among the initial responses of seedlings to water stress.     

Photoinhibition and Recovery of Photosynthesis in Canker-susceptible and Resistant Needles of Cypress (Cupressus sempervirens L.)

The aim of the present investigation was to test the hypothesis that the cypress canker caused by a fungus (Seiridium cardinale) infection induced effects on photosynthesis which could be related to photoinhibition and the process of recovery in susceptible and resistant needles. Photoinhibition of photosynthesis and recovery was studied in canker‐infected susceptible and resistant needles of cypress (Cupressus sempervirens L.) under controlled conditions (irradiation of detached needles to approximately 1900 μmol/m²/s). The degree of photoinhibition was determined by means of the ratio of variable to maximum chlorophyll (Chl) fluorescence (F_v/F_m) and electron transport measurements. The potential efficiency of photosystem (PS) II, F_v/F_m declined, and F_o increased significantly in canker‐susceptible needles, while F_o did not change in resistant needles. In isolated thylakoids, high light (HL) decreased the rate of whole chain and PS II activity markedly more in susceptible than in resistant needles. A smaller reduction of PS I activity was noticed only in susceptible needles. Upon subsequent dark incubation, fast recovery was noticed in both needle types and reached maximum rates of PS II efficiency similar to those noticed in non‐photoinhibited needles. The artificial exogenous electron donors such as diphenyl carbazide (DPC), NH₂OH and Mn²⁺ failed to restore the HL induced loss of PS II activity in susceptible needles, while DPC and NH₂OH significantly restored it in resistant needles. The results suggest that HL inactivates the donor side of PS II in resistant and the acceptor side of PS II in susceptible needles. The results on the quantification of the PS II reaction centre protein D1 and 33 kDa protein of water‐splitting complex following HL exposure showed pronounced differences between susceptible and resistant needles. The marked loss of PS II activity in HL‐irradiated needles was due to the marked loss of D1 protein in susceptible and 33 kDa protein in resistant needles, respectively.     

The xanthophyll cycle and acclimation of Pinus ponderosa and Malva neglecta to winter stress

Seasonal differences in the efficiency of open PSII units (F _v/F _m), leaf pigment composition and xanthophyll cycle conversion (Z+A)/(V+A+Z), leaf adenylate status, and photosynthetic capacity were investigated in Pinus ponderosa (Ponderosa pine) and Malva neglecta. In P. ponderosa, acclimation to winter involved a lower photosynthetic capacity, higher carotenoid to chlorophyll ratio, persistent reductions in F _v/F _m corresponding to persistent retention of Z+A, and no change in foliar ATP/ADP ratios. In contrast, M. neglecta characterized in winter exhibited higher rates of photosynthesis than in summer with no change in carotenoid to chlorophyll ratio, while small nocturnally persistent reductions in F _v/F _m were observed exclusively on colder winter nights when nocturnal retention of Z+A, and high ATP/ADP ratios were also present. Upon removal of winter-stressed leaves or needles from the field to room temperature, a portion of F _v/F _m relaxed within 15 min of warming and recovery was completed within 5 h in M. neglecta but required 100 h in P. ponderosa. In M. neglecta, the entire recovery of F _v/F _m correlated with decreases in the foliar ATP/ADP ratio, while in P. ponderosa this ratio remained unchanged. Possible ATP-dependent forms of sustained (Z+A)-dependent energy dissipation are discussed including a nocturnally retained pH gradient on cold winter nights. The slow recovery in pine involved not only retention of Z+A, but apparently also a persistent engagement of Z+A for energy dissipation via an unidentified mechanism. Received: 15 May 1998 / Accepted: 9 November 1998     

High Irradiance Induced Changes of Photosystem 2 in Young and Mature Needles of Cypress (Cupressus sempervirens L.)

Photoinhibition of photosynthesis was studied in young and mature detached sun needles of cypress under high irradiance (HI) of about 1 900 mol m^–2 s^–1. The degree of photoinhibition was determined by means of the ratio of variable to maximum chlorophyll (Chl) fluorescence (F_v/F_m) and electron transport measurements. Compared with the mature needles, the young needles, containing about half the amount of Chl a+b per unit area, exhibited a higher proportion of total carotenoids (Car) as xanthophyll cycle pigments and had an increased ratio of Car/Chl a+b. The potential efficiency of photosystem (PS) 2, F_v/F_m, markedly declined in HI-treated young needles without significant increase of F₀ level. In contrast, the F_v/F_m ratio declined with significant increase of F₀ level in mature needles. In isolated thylakoids, the rate of whole chain and PS2 activity markedly decreased in young HI-needles in comparison with mature needles. A smaller inhibition of PS1 activity was observed in both needles. In the subsequent dark incubation, fast recovery was found in both needle Types that reached maximum PS2 efficiencies similar to those observed in non-photoinhibited needles. The artificial exogenous electron donors DPC, NH₂OH, and Mn²⁺ failed to restore the HI-induced loss of PS2 activity in mature needles, while DPC and NH₂OH significantly restored it in young needles. Hence, HI-inactivation was on the donor side of PS2 in young needles and on the acceptor side of PS2 in mature needles. Quantification of the PS2 reaction centre proteins D1 and 33 kDa protein of water splitting complex following HI-exposure of needles showed pronounced differences between young and mature needles. The large loss of PS2 activity in HI-needles was due to the marked loss of D1 protein of the PS2 reaction centre in mature needles and of the 33 kDa protein in young needles.     

Persistent effects of low concentrations of SO2 and NO2 on photosynthesis in Scots pine (Pinus sylvestris) needles

In an open-field experiment, 50-year-old trees of Scots pine (Pinus sylvestris L.) were fumigated with low concentrations of SO₂ and NO₂ (10–15 nl I^?1) during the growing season in four consecutive years (1988 to 1991). Results from the autumn and early winter of 1991 and 1992 are presented. The maximum photochemical efficiency of photosystem II (PSII), as indicated by the ratio of variable to maximum fluorescence (F_v/F_M) was assessed in current and one-year-old needles from the top and the bottom of the canopy. Furthermore, simultaneous measurements of photosynthetic O₂ evolution and chlorophyll fluorescence were made in current-year needles at 20°C. In general, the F_v/F_M ratio as well as the gross rate of O₂ evolution in needles of fumigated trees was not significantly different from that in needles of control trees during the fumigation period. However, both current and one-year-old needles sampled in November and December 1991 from the top of the canopy of fumigated trees had significantly lower F_v/F_M values than corresponding needles of control trees. Similar differences in F_v/F_M correlated with the treatments were observed in needles from the bottom of the canopy, indicating that the depression of F_v/F_M in needles of fumigated trees was not due to an increased susceptibility to photoinhibition. In 1992, when no fumigation occurred, differences in F_v/F_M between the treatments were not significant during autumn and early winter. The gross rate of O₂ evolution at high irradiances was significantly lower in current-year needles of fumigated trees sampled in November and December 1991 than in those of control trees. Furthermore, a nearly identical linear relationship between the quantum yield of PSII electron transport determined from chlorophyll fluorescence and the quantum yield of O₂ evolution (gross rate of O₂ evolution/PPFD) was found during autumn and early winter. This appeared to be largely a result of changes in the thermal energy dissipation within PSII. The observed differences in photosynthetic characteristics correlated with the different treatments after the fumigation period is suggested to be mainly caused by increased sensitivity of the needles of fumigated trees to low and subfreezing temperatures. However, current-year needles of fumigated trees tended to have a lower N content than those of control trees, which may partly explain the differences in gross photosynthesis between fumigated and control trees.     

Species-specific acclimation to strong shade modifies susceptibility of conifers to photoinhibition

Photoinhibitory processes in the photosynthetic apparatus of the seedlings of Abies alba (Mill.), Picea abies (Karst.), and Pinus mugo (Turra) growing under strong shade (5 % of full solar irradiance) or full irradiance conditions were investigated in winter and spring using chlorophyll a fluorescence techniques. The extent of photoinhibition in needles as indicated by a decrease in maximum quantum yield of PS II photochemistry (F_v/F_m) depended on species, air temperature and acclimation to the light environment. Unexpectedly, shade-tolerant Abies alba was less affected by low-temperature photoinhibition compared to the other species. F_v/F_m recovered with increasing air temperature. During winter, the seedlings of Picea abies growing in shade showed higher F_v/F_m than those from full light. Non-photochemical quenching of fluorescence (NPQ) measured at the same levels of actinic light was higher in needles acclimated to full light except for Abies alba in February. Photosynthetic performance in term of ETR (apparent electron transfer rate) was also higher in full light-acclimated needles. In April, at ambient temperature, recovery of PS II efficiency from the stress induced by illumination with saturating light was faster in the needles of Picea abies than in those of Abies alba. The shade-acclimated needles of Abies alba and Picea abies showed greater down-regulation of PS II induced by high light stress.     

Seasonal Effects on Photosynthetic Electron Transport and Fluorescence Properties in Isolated Chloroplasts of Pinus silvestris

Chloroplasts were isolated from primary needles of 1-year-old seedlings and from secondary needles of a 20-year-old pine tree in a natural stand. In autumn the electron transport capacities of PSII, PSI and PS (II + I) decreased and the electron transport between PSII and PSI became inhibited in October in the 20-year-old tree. This inhibition lasted until May the following year. The partial reactions of PSI and PSII still showed low but fairly constant rates during the whole winter seedlings. Seasonal changes in the electron transport properties of 1-year-old showed the same general trends as observed in the 20-year-old tree, but the changes were less pronounced. However, in snow-covered seedlings the PSI-mediated electron transport and the electron transport from H₂O to NADP increased during the late winter when the seedlings were still covered by snow. The total chlorophyll content of the needles decreased in autumn and winter. Low temperature fluorescence ratios of F692/F680 and F726/F680 indicated more severe destruction of the chlorophyll a antennae closely associated with the two photosystems than of the light harvesting chlorophyll a/b complex. In this case, too, the changes were more pronounced in the 20-year-old tree than in the 1-year-old seedlings. The chlorophyll/P700 ratios indicated a more marked reduction in the reaction centre molecules during autumn than in the antennae chlorophyll molecules. The changes in electron transport and low temperature fluorescence properties which occurred during autumn and winter were mainly reversed during spring.     

Effects of soil warming during spring on photosynthetic recovery in boreal Norway spruce stands

The effect of soil thawing and soil temperature on postwinter recovery of photosynthetic capacity was studied, during late spring and early summer, in Norway spruce stands in northern Sweden. Soil temperature was manipulated by means of buried heating cables. The warming treatment was applied to stands with low (natural) and high (fertilized) availability of nutrients. Soil thawing, expressed as water availability, was followed by means of sapflow in stems, and shoot water potentials. The recovery of photosynthetic capacity was assessed by measuring the rate of light-saturated photosynthesis (A_max), and maximum photochemical efficiency of photosystem II in detached shoots, and chlorophyll a fluorescence. Accumulation of starch reserves in the needles was followed as an independent indicator of photosynthetic performance in situ. Snowmelt and soil thawing occurred more than one month earlier in heated than in unheated plots. This was expressed both as sapflow and as differences in shoot water potential between treatments. During May, the rates of A_max were significantly higher on heated than on control plots. The effect of soil warming on A_max was, however, not reflected in chlorophyll fluorescence or needle starch content. The time course of the recovery of photosynthetic capacity was mainly controlled by mean air temperature and by the frequency of severe night frosts, and to a lesser extent by earlier soil thawing and higher soil temperatures.     

Inhibition of photosynthesis in current-year needles of unfertilized and fertilized Norway spruce [Picea abies (L.) Karst.] during autumn and early winter

Inhibition of photosynthesis was followed during autumn and early winter in current-year sun and shade needles of unfertilized and fertilized Norway spruce [Picea abies (L.) Karst.] by simultaneous measurements of photosynthetic O₂ evolution and chlorophyll a fluorescence at 20 °C. The CO₂-saturated rate of O₂ evolution was generally higher in sun needles of fertilized trees than in those of unfertilized trees over a wide range of incident photon flux densities (PFDs). Furthermore, the maximum photo-chemical efficiency of photosystem (PS) II, as indicated by the ratio of variable to maximum fluorescence (F_V/F_M) was generally higher for sun needles of fertilized trees. The depression of f_v/f_m during frost periods was more pronounced in sun needles than in shade needles, indicating that winter inhibition in Norway spruce is strongly light-dependent. However, the inhibition of the rate of O₂ evolution at high PFDs in needles of fertilized trees during early winter was partly independent of the light regime experienced by those needles in the field, which appeared to result in a pronounced decrease in the proportion of oxidized PS II reaction centres in shade needles. A nearly identical linear relationship between the quantum yield of PS II electron transport determined by chlorophyll fluorescence and the quantum yield of O₂ evolution (gross rate of O₂ evolution/PFD) was obtained for the investigated types of needles during autumn and early winter. Except for shade needles of fertilized trees, this appeared to be largely achieved by adjustments in thermal energy dissipation within PS II.     

Characterization of the photosynthetic apparatus in cortical bark chlorenchyma of Scots pine

Winter-induced inhibition of photosynthesis in Scots pine (Pinus sylvestris L.) needles is accompanied by a 65% reduction of the maximum photochemical efficiency of photosystem II (PSII), measured as F _v/F _m, but relatively stable photosystem I (PSI) activity. In contrast, the photochemical efficiency of PSII in bark chlorenchyma of Scots pine twigs was shown to be well preserved, while PSI capacity was severely decreased. Low-temperature (77 K) chlorophyll fluorescence measurements also revealed lower relative fluorescence intensity emitted from PSI in bark chlorenchyma compared to needles regardless of the growing season. Nondenaturating SDS-PAGE analysis of the chlorophyll–protein complexes also revealed much lower abundance of LHCI and the CPI band related to light harvesting and the core complex of PSI, respectively, in bark chlorenchyma. These changes were associated with a 38% reduction in the total amount of chlorophyll in the bark chlorenchyma relative to winter needles, but the Chl a/b ratio and carotenoid composition were similar in the two tissues. As distinct from winter pine needles exhibiting ATP/ADP ratio of 11.3, the total adenylate content in winter bark chlorenchyma was 2.5-fold higher and the estimated ATP/ADP ratio was 20.7. The photochemical efficiency of PSII in needles attached to the twig recovered significantly faster (28–30 h) then in detached needles. Fluorescence quenching analysis revealed a high reduction state of Q _A and the PQ-pool in the green bark tissue. The role of bark chlorenchyma and its photochemical performance during the recovery of photosynthesis from winter stress in Scots pine is discussed.     

Photosynthetic changes that occur during aging of cypress (Cupressus sempervirens L.) needles

2-years-old cypress needles (A2) were physiologically most active with regard to net photosynthetic (P _N) and electron transport rates. Variable to maximum fluorescence (F_v/F_m) ratios of dark-adapted needles were higher in A2 needles than in current year (A1) or senescent (A4) needles. Lower F_v/F_m values in these stages seemed to be caused not by photoinhibition but by a low photochemical capacity as suggested from the chlorophyll (Chl) a/b ratios. In isolated thylakoids, lower rates of whole chain and photosystem 2 (PS2) activities were observed in A4 needles, while higher rates were observed in A2 needles. A similar trend was noticed for contents of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPC) and total soluble proteins. The artificial exogenous electron donor Mn²⁺ failed to restore the loss of PS2 activity in 3-year-old (A3) and A4 needles, while diphenylcarbazide and NH₂OH significantly restored the loss of PS2 activity. The marked loss of PS2 activity in A4 needles was primarily the result of the loss of 33, 28–25, 23, and 17 kDa polypeptides. A marked loss of RuBPC activity in A4 needles is mainly due to the loss of 15 (SSU) and 55 (LSU) kDa polypeptides.     

Distribution of N, Rubisco and photosynthesis in Pinus pinaster and acclimation to light

The light–nitrogen hypothesis suggests canopy photosynthesis is maximized when there is a positive relationship between irradiance received by foliage, its nitrogen content (per unit area N_area), and maximum rate of photosynthesis (A_max). Relationships among relative irradiance and N_area, allocation of nitrogen within the photosynthetic apparatus to Rubisco and chlorophyll, and A_max were examined in Pinus pinaster Ait. needles up to 6 years of age. Measurements were made before bud break in August 1998, and in May 1999 after the first ‘winter’ rains. In August, N_area in P. pinaster needles decreased from 5·1 to 5·7 g m^?2 in sunlit 1‐year‐old needles to 2·3 g m^?2 in shaded 6‐year‐old needles. In May, N_area was 5–40% less but spatial trends were the same. At both sampling dates, A_max was less in old shaded needles compared with young sunlit needles, and was thus consistent with the light–nitrogen hypothesis. Relationships between N_area and A_max were positive at both dates yet varied in strength and form. Allocation of nitrogen within the photosynthetic apparatus was qualitatively consistent with acclimation to light (i.e. Rubisco/Chl decreased with shading), but quantitatively suboptimal with respect to photosynthesis owing to consistent over‐investment in Rubisco. This over‐investment increased with height in the canopy and was greater in May than in August.     

Violaxanthin cycle pigment de-epoxidation and thermal dissipation of light energy in three boreal species of evergreen conifer plants

We studied carotenoid composition and chlorophyll fluorescence in two-year-old needles from Siberian spruce (Picea obovata (L.) Karst.), Siberian fir (Abies sibirica L.), and common juniper (Juniperus communis L.). The highest values of maximum PSII photochemical activity (F _v/F _m) equaling 0.82–0.85 were observed in July–September. The decrease in F _v/F _m in December–March was more pronounced in juniper (down to 0.15) than in spruce and fir (0.45–0.50). In May, we observed a nearly complete recovery in maximum PSII photochemical activity in fir and spruce (0.72–0.77), while in juniper, the F _v/F _m value was notably lower (0.65–0.67). The amount of thermal dissipation of energy absorbed by PSII LHC did not exceed 30% in summer and equaled 60–90% in winter and early spring. The carotenoid pool consisted mainly of xanthophylls, among which lutein (70%), neoxanthin (7–10%), and a violaxanthin cycle (VXC) component — violaxanthin (3–15%) were constantly present. The accumulation of two other VXC pigments—zeaxanthin and antheraxanthin, was noted in December–March. In July, these xanthophylls were not identified. We discovered a direct connection between VXC pigment de-epoxidation level and light energy thermal dissipation in boreal conifer leaves. Such association reflects the non-species-specific character of the mechanism for quenching zeaxanthin-dependent nonphotochemical chlorophyll fluorescence in PSII LHC in winter and spring.     

Salt stress affects water relations,photosynthesis, and oxidative defense mechanisms in Solanum melongena L.

Abstract

A greenhouse experiment was conducted to examine the salt-induced changes in some key physio-biochemical attributes in eggplant (cv. New Noble) plants. Eggplant plants were grown under varying levels (0, 50, 100, and 150 mM) of sodium chloride under greenhouse conditions supplied with natural light and other climatic conditions. Varying saline regimes in growth medium significantly reduced the shoot and root fresh and dry weights, shoot and root lengths, relative water content, chlorophyll a and b pigments, photosynthetic rate (A), water-use efficiency, stomatal conductance (g _s ), leaf and root K⁺, total phenolics, total soluble proteins, activity of superoxide dismutase (SOD), and leaf water and osmotic potentials in all eggplant plants. However, in contrast, saline regimes of the root growing medium did not affect transpiration rate (E), internal CO₂ concentration (C _i ), C _i /C _a ratio, photochemical quenching (qP), non-photochemical quenching, efficiency of photosystem-II (F _v /F _m ), leaf and root Ca²⁺ as well as ascorbic acid (AsA) contents in eggplant. A significant increase was observed in leaf turgor potential, free proline and glycinebetaine contents, leaf and root Na⁺ contents, malondialdehyde and hydrogen peroxide (H₂O₂) contents and activities of peroxidase (POD) and catalase (CAT) in eggplant plants under varying saline regimes. Overall, salt-induced growth reduction in eggplant plants was found to be associated with high accumulation of Na⁺ in both roots and shoots, which adversely affected photosynthetic capacity, chlorophyll pigments, K⁺ and Ca²⁺ contents, H₂O₂ and AsA levels and activities of SOD, POD, and CAT.     

Different Responses of Norway Spruce Needles from Shaded and Exposed Crown Layers to the Prolonged Exposure to Elevated CO2 Studied by Various Chlorophyll a Fluorescence Techniques

The acclimation depression of capacity of photon utilisation in photochemical reactions of photosystem 2 (PS2) can develop already after three months of cultivation of the Norway spruces (Picea abies [L.] Karst.) under elevated concentrations of CO₂ (i.e., ambient, AC, + 350 µmol(CO₂) mol^–1 = EC) in glass domes with adjustable windows. To examine the role that duration of EC plays in acclimation response, we determined pigment contents, rate of photosynthesis, and parameters of chlorophyll a fluorescence for sun and shade needles after three seasons of EC exposure. We found responses of shaded and exposed needles to EC. Whereas the shaded needles still profited from the EC and revealed stimulated electron transport, for the exposed needles the stimulation of both electron transport activity and irradiance saturated rate of CO₂ assimilation (P _Nmax) under EC already disappeared. No signs of the PS2 impairment were observed as judged from high values of potential quantum yield of PS2 photochemistry (F_V/F_M) and uniform kinetics of Q_A reoxidation for all variants. Therefore, the long-term acclimation of the sun-exposed needles to EC is not necessarily accompanied with the damage to the PS2 reaction centres. The eco-physiological significance of the reported differentiation between the responses of shaded and sun exposed needles to prolonged EC may be in changed contribution of the upper and lower crown layers to the production activity of the tree. Whereas for the AC spruces, P _Nmax of shaded needles was only less than 25 % compared to exposed ones, for the EC spruces the P _Nmax of shaded needles reached nearly 40 % of that estimated for the exposed ones. Thus, the lower shaded part of the crown may become an effective consumer of CO₂.     

Winter photoinhibition in needles of <Emphasis Type="Italic">Taxus baccata</Emphasis> seedlings acclimated to different light levels

Seasonal variability of maximum quantum yield of PSII photochemistry (F_v/F_m) was studied in needles of Taxus baccata seedlings acclimated to full light (HL, 100% solar irradiance), medium light (ML, 18% irradiance) or low light (LL, 5% irradiance). In HL plants, F_v/F_m was below 0.8 (i.e. state of photoinhibition) throughout the whole experimental period from November to May, with the greatest decline in January and February (when F_v/F_m value reached 0.37). In ML seedlings, significant declines of F_v/F_m occurred in January (with the lowest level at 0.666), whereas the decline in LL seedlings (down to 0.750) was not significant. Full recovery of F_v/F_m in HL seedlings was delayed until the end of May, in contrast to ML and LL seedlings. F_v/F_m was significantly correlated with daily mean (T _mean), maximal (T _max) and minimal (T _min) temperature and T _min was consistently the best predictor of F_v/F_m in HL and ML needles. Temperature averages obtained over 3 or 5 days prior to measurement were better predictors of F_v/F_m than 1- or 30-day averages. Thus our results indicate a strong light-dependent seasonal photoinhibition in needles of T. baccata as well as suggest a coupling of F_v/F_m to cumulative temperature from several preceding days. The dependence of sustained winter photoinhibition on light level to which the plants are acclimated was further demonstrated when plants from the three light environments were exposed to full daylight over single days in December, February and April and F_v/F_m was followed throughout the day to determine residual sensitivity of electron transport to ambient irradiance. In February, the treatment revealed a considerable midday increase in photoinhibition in ML plants, much less in HL (already downregulated) and none in LL plants. This suggested a greater capacity for photosynthetic utilization of electrons in LL plants and a readiness for rapid induction of photoinhibition in ML plants. Further differences between plants acclimated to contrasting light regimes were revealed during springtime de-acclimation, when short term regeneration dynamics of F_v/F_m and the relaxation of nonphotochemical quenching (NPQ) indicated a stronger persistent thermal mechanism for energy dissipation in HL plants. The ability of Taxus baccata to sustain winter photoinhibition from autumn until late spring can be beneficial for protection against an excessive light occurring together with frosts but may also restrict photosynthetic carbon gain by this shade-tolerant species when growing in well illuminated sites.     

Development of photoautotrophy and photoinhibition of Gardenia jasminoides plantlets during micropropagation

This paper reports on the fast fluorescence responses of Gardenia jasminoides Ellis plantlets, at two successive stages (shoot multiplication and root induction) of culture in vitro. We test whether plantlets in vitro suffer photoinhibition during culture and whether the degree of photoautotrophy of these mixotrophic plantlets has any effect on the extent of photoinhibitory impairment. In this regard the effects of different sucrose levels in the medium and PPFD during growth on the development of photoautotrophy and the extent of photoinhibition were evaluated. Plantlets were grown under low, intermediate, and high (50, 100, and 300 mol m^-2 s^-1) PPFD, and at 3 different sucrose concentrations (0.5, 1.5, and 3.0%, w/v) in the medium, during shoot multiplication. During root induction the same growth conditions were assayed except for the high PPFD. The development of photoautotrophy was assessed via the difference between the stable carbon isotope composition of sucrose used as heterotrophic carbon source and that of leaflets grown in vitro. Plantlets from root induction showed more developed photoautotrophy than those from shoot multiplication. For both stages the low-sucrose medium stimulated the photoautotrophy of plantlets in vitro. In addition, intermediate PPFD induced photoautotrophy during shoot multiplication. For plantlets of both culture stages at the lowest PPFD no photoinhibition occurred irrespective of the sucrose concentration in media. However, during the shoot multiplication stage chlorophyll fluorescence measurements showed a decrease in F _v /F _m and in t _1/2 as growing PPFD increased, indicating photoinhibitory damage. The decline of F _v /F _m was caused mostly by an increase in F _o , indicating the inactivation of PSII reaction centers. However plantlets growing under low sucrose showed reduced susceptibility to photoinhibition. During root induction, only plantlets cultured with high sucrose showed a decrease in F _v /F _m as PPFD increased, although t _1/2 remained unchanged. In this case, the decline of F _v /F _m was mostly due to a decrease in F _m , which indicates increased photoprotection rather than occurrence of photodamage. Therefore, growth in low-sucrose media had a protective effect on the resistance of PSII to light stress. In addition, plantlets were more resistant to photoinhibition during root induction than during shoot multiplication. Results suggest that increased photoautotrophy of plantlets reduces susceptibility to photoinhibition during gardenia culture in vitro.Abbreviations AP apparent photosynthesis - Chl total chlorophyll content - Chl a/b chlorophyll a-to-b ratio - Chl/Car total chlorophyll-to-carotenoids ratio - ¹³C ratio of ¹³C/¹²C relative to PeeDee belemnite standard - F _m maximum chlorophyll fluorescence - F _o fluorescence emission when all reaction centres are open and the photochemical quenching is minimal - F _v variable chlorophyll fluorescence (F _m -F _o ) - F _v /F _m the ratio of variable to maximum chlorophyll fluorescence, indicator photochemical efficiency of PSII - MS medium Murashige and Skoog (1962) medium - PPFD photosynthetic photon flux density - Rd dark respiration, t _1/2 the half-time of the increase from F _o to F _m - IAA indole butyric acid     

Response of tomato plants to chilling stress in association with nutrient or phosphorus starvation

The experiments were conducted on two tomato cultivars: Garbo and Robin. Mineral starvation due to plant growth in 20-fold diluted nutrient solution (DNS) combined with chilling reduced the rate of photosynthesis (P _N) and stomatal conductance (g) to a greater extent than in plants grown in full nutrient solution (FNS). In phosphate-starved tomato plants the P _N rate and stomatal conductance decreased more after chilling than in plants grown on FNS. In low-P plants even 2 days after chilling the recovery of CO₂ assimilation rate and stomatal conductance was low. A resupply of phosphorus to low-P plants (low P + P) did not improve the rate of photosynthesis in non-chilled plants (NCh) but prevented P_N inhibition in chilled (Ch) plants. The greatest effect of P resupply was expressed as a better recovery of photosynthesis and stomatal conductance, especially in non-chilled low P + P plants. The F _v/F _m (ratio of variable to maximal chlorophyll fluorescence) decreased more during P starvation than as an effect of chilling. Supplying phosphorus to low-P plants caused the slight increase in the F _v/F _mratio. In conclusion, after a short-term chilling in darkness a much more drastic inhibition of photosynthesis was observed in nutrient-starved or P-insufficient tomato plants than in plants from FNS. This inhibition was caused by the decrease in both photochemical efficiency of photosystems and the reduction of stomatal conductance. The presented results support the hypothesis that tomato plants with limited supply of mineral nutrients or phosphorus are more susceptible to chilling.     

Leaf age as a factor in anatomical and physiological acclimative responses of Taxus baccata L. needles to contrasting irradiance environments

To evaluate the acclimative ability of current-year and previous-year needles of a shade tolerant conifer Taxus baccata L. to contrasting irradiance conditions, seedlings were raised under 27% solar irradiance and at 3 years of age they were transferred to an experimental garden and grown for one season under full irradiance (HL), 18% irradiance (ML) or 5% irradiance (LL). Whereas previous year needles did not change anatomically, current year needles in HL were thicker and had a thicker palisade and spongy mesophyll, and greater leaf mass per area than ML or LL needles. LL needles had greater nitrogen concentration than HL needles irrespective of age but only previous year LL needles also had an increased N per area content, thanks to their lack of reduction in LMA. Adjustment of chlorophyll and carotenoid content occurred in both needle age classes with LL and ML needles having much higher concentrations but, in current year needles, only slightly higher per area content than HL needles. Chlorophyll a/b ratio was not affected by age or irradiance. These modifications had no significant effect on photosynthetic capacities, which did not significantly differ between the age classes in HL or LL treatment and between treatments. On the other hand, high growth irradiance resulted in a greater photochemical yield, photochemical quenching, apparent electron transport rate and inducible non-photochemical quenching in needles formed in the current season. In previous year needles, however, only inducible NPQ was enhanced by high irradiance with other parameters remaining identical among treatments. To test sensitivity to photoinhibition, at the end of the summer plants from the three irradiance levels were transferred to a HL situation and F _v/F _M was determined over the following 18 days. Sensitivity to photoinhibition was negatively related to growth irradiance and previous year needles were less photoinhibited than current year needles. Thus, differences in acclimation ability between needle age classes were most pronounced at the level of anatomy and light reactions of photosynthesis, both of which showed almost no plasticity in previous year needles but were considerably modified by irradiance in current year needles.     

Close relationship between the state of the xanthophyll cycle pigments and photosystem II efficiency during recovery from winter stress

The potential involvement of the xanthophyll cycle in photoprotection of overwintering evergreen plants was investigated. Leaves from five evergreen species. Pseudotsuga menziesii, Pinus panderosa, Euonyums kiautschovicus. Mahonia repens and Malva neglecta, were collected from the field predawn during winter and transferred to the laboratory where chlorophyll fluorescence emission as well as the chlorophyll and carotenoid composition were ascertained periodically for 4.5 days. Leaves and needles from all species were found to have retained large amounts of the xanthophyll cycle pigments zeaxanthin and antheraxanthin, and they exhibited sustained low values of the intrinsic efficiency of photosystem II (PSII; measured as the ratio of variable to maximal fluorescence, F_v/F_m) upon collection. The increase in PSII efficiency was biphasic, with a rapid phase (requiring several hours) and a slow phase (requiring several days). Changes in the conversion state of the xanthophyll cycle were found to correlate with increases in PSII efficiency in both phases, with the latter phase involving large increases in both F_m (maximal fluorescence) and F_o (minimal fluorescence) throughout the period of recovery. The relationship between F_m quenching (expressed as nonphotochemical or Stern-Volmer quenching [NPQ] of F_m, i.e. F_m/ F_m–1) and F_o quenching (F_o/F_o–1) was linear, as expected for changes in xanthophyll cycle-dependent energy dissipation in the antenna complexes. Furthermore, the relationship between F_v/F_m and NPQ during recovery followed the theoretical relationship predicted for changes in the rate constant for energy dissipation in the antenna complexes. This fit between the theoretical relationship and the actual data indicates that all changes in NPQ or F_v/F_m can be accounted for by changes in this rate constant. The results suggest a role for the photoprotective xanthophyll cycle-dependent dissipation process in the lowered efficiency of PSII observed in coldstressed evergreen plants in the field.