Enhancement of susceptivity to photoinhibition and photooxidation in rice chlorophyll <Emphasis Type="Italic">b</Emphasis>-less mutants

Two rice chlorophyll (Chl) b-less mutants (VG28-1, VG30-5) and the respective wild type (WT) plant (cv. Zhonghua No. 11) were analyzed for the changes in Chl fluorescence parameters, xanthophyll cycle pool, and its de-epoxidation state under exposure to strong irradiance, SI (1 700 μmol m⁻² s⁻¹). We also examined alterations in the chloroplast ultrastructure of the mutants induced by methyl viologen (MV) photooxidation. During HI (0–3.5 h), the photoinactivation of photosystem 2 (PS2) appeared earlier and more severely in Chl b-less mutants than in the WT. The decreases in maximal photochemical efficiency of PS2 in the dark (F_v/F_m), quantum efficiency of PS2 electron transport (Φ_PS2), photochemical quenching (q_P), as well as rate of photochemistry (P_rate), and the increases in de-epoxidation state (DES) and rate of thermal dissipation of excitation energy (D_rate) were significantly greater in Chl b-mutants compared with the WT plant. A relatively larger xanthophyll pool and 78–83 % conversion of violaxanthin into antheraxanthin and zeaxanthin in the mutants after 3.5 h of HI was accompanied with a high ratio of inactive/total PS2 (0.55–0.73) and high 1–q_P (0.57–0.68) which showed that the activities of the xanthophyll cycle were probably insufficient to protect the photosynthetic apparatus against photoinhibition. No apparent difference of chloroplast ultrastructure was found between Chl b-less mutants and WT plants grown under low, LI (180 μmol m⁻² s⁻¹) and high, HI (700 μmol m⁻² s⁻¹) irradiance. However, swollen chloroplasts and slight dilation of thylakoids occurred in both mutants and the WT grown under LI followed by MV treatment. These typical symptoms of photooxidative damage were aggravated as plants were exposed to HI. Distorted and loose scattered thylakoids were observed in particular in the Chl b-less mutants. A greater extent of photoinhibition and photooxidation in these mutants indicated that the susceptibility to HI and oxidative stresses was enhanced in the photosynthetic apparatus without Chl b most likely as a consequence of a smaller antenna size.     

The interacting effect of urea and fenoxaprop-P-ethyl on photosynthesis and chlorophyll fluorescence in Perilla frutescens

We studied the effect of herbicide and nitrogen supply on photosynthesis in Perilla frutescens L. Britt. Plants were exposed to combined treatment of urea and herbicide, fenoxaprop-P-ethyl (FPE), in various concentrations. FPE reduced significantly chlorophyll (Chl) content, photosynthetic rate, and stomatal conductance, but increased significantly intercellular CO₂ concentration; thus, FPE inhibited significantly the photosynthetic capacity. In addition, FPE also decreased significantly the PSII photochemical efficiency, effective quantum yield of photochemical energy conversion in PSII, PSII potential activity, and photochemical quenching of variable Chl fluorescence. It also decreased nonphotochemical quenching. It indicated that FPE impaired PSII and blocked the electron transport in light reaction. The urea treatment at moderate concentration (1–4 g L^?1) could antagonize the negative effect of FPE, while the high urea concentration (8 g L^?1) aggravated this effect. The treatment with urea (4 g L^?1) and then with FPE (1.33 mL L^?1) enhanced Chl content index, photosynthetic rate, and stomatal conductance by 12.5, 36.1, and 28.5% compared to FPE treatment alone. Thus, we suggested to treat plants first with urea (4 g L^?1) and then by FPE (1.33 mL L^?1) as the best and the safest method to balance the fertilization and weeding.     

Photosynthetic characteristics and antioxidant enzyme system in high-chlorophyll rice Gc mutant

The physiological photosynthetic characteristics and antioxidant enzyme system of the high-chlorophyll rice (Oryza sativa L.) mutant (Gc) and its wild type (Zhenshan 97B) were compared and analyzed. Resulting data showed that the total chlorophyll (Chl) and Chl b contents in the Gc mutant were significantly increased by 19.0 and 81.7%, respectively, while the increase in Chl a and thylakoid membrane protein contents was insignificant. The net photosynthetic rate (P _N) was significantly higher in the mutant; stomatal conductance, intercellular CO₂ concentration, and transpiration rate decreased significantly, and water-use efficiency increased significantly, indicating the higher photochemical efficiency of the mutant. The chlorophyll fluorescence parameters: electron transport rate and effective quantum yield of PSII photochemistry of the mutant were significantly higher than those of Zhenshan 97B. The nonphotochemical quenching of the mutant under light adaptation increased by 52.3%. The enzymatic activity of superoxide dismutase, peroxidas, and catalase in the mutant roots and leaves were all higher than those for the wild-type plants. It is believed that the higher activity of antioxidant enzymes in the mutant may be an important factor making difficult the photo-inactivation of Chl, and thus, increasing the content of Chl, especially Chl b.     

Epidermal UV-shielding and photosystem II adjustment in wild type and chlorina f2 mutant of barley during exposure to increased PAR and UV radiation

The wild-type barley (WT; Hordeum vulgare L.) and its chlorophyll (Chl) b-less mutant chlorina f2 (clo f2) grown under shaded conditions in a greenhouse were transferred to outdoor conditions in early June with predominantly bright sunny days. During 6 days following transfer of plants we monitored the content of photosynthetic pigments, functional state of photosystem II (PSII) by means of Chl fluorescence induction kinetics and epidermal UV-shielding efficiency using Chl fluorescence imaging technique. Clo f2 mutant was more sensitive to exposure to an enhanced natural solar irradiance than WT barley. Nevertheless, clo f2 as well as WT were able to cope with stressful outdoor conditions, as was documented by the recovery of Chl a content and the maximal photochemical efficiency of PSII (F_V/F_M) after an initial decline. This was due to the immediate carotenoid-mediated photoprotection, reflected by strongly increased total carotenoids content and thermal energy dissipation localized within light-harvesting complexes of PSII (assessed by non-photochemical quenching of minimal fluorescence level). The positive acclimation response was further documented by an enhanced light-saturated electron transport rate through PSII (ETR). Based on the ratios of blue- to UV-excited Chl fluorescence we found that for both WT and clo f2 epidermal UV-shielding increased clearly after transfer to outdoor conditions and reached a saturation level after 3 days. In comparison with WT, clo f2 exhibited lower ability to induce UV-shielding. The kinetics of UV-shielding development during the outdoor treatment was different for the particular leaf regions. We suggest that this is related to the different age and developmental stage of the tissue along the leaf blade. The complementarity of carotenoid-mediated photoprotection and UV-shielding in acclimation of the assimilatory apparatus to increased visible and UV radiation is discussed.     

A chlorophyll-deficient rice mutant with impaired chlorophyllide esterification in chlorophyll biosynthesis

Chlorophyll (Chl) synthase catalyzes esterification of chlorophyllide to complete the last step of Chl biosynthesis. Although the Chl synthases and the corresponding genes from various organisms have been well characterized, Chl synthase mutants have not yet been reported in higher plants. In this study, a rice (Oryza Sativa) Chl-deficient mutant, yellow-green leaf1 (ygl1), was isolated, which showed yellow-green leaves in young plants with decreased Chl synthesis, increased level of tetrapyrrole intermediates, and delayed chloroplast development. Genetic analysis demonstrated that the phenotype of ygl1 was caused by a recessive mutation in a nuclear gene. The ygl1 locus was mapped to chromosome 5 and isolated by map-based cloning. Sequence analysis revealed that it encodes the Chl synthase and its identity was verified by transgenic complementation. A missense mutation was found in a highly conserved residue of YGL1 in the ygl1 mutant, resulting in reduction of the enzymatic activity. YGL1 is constitutively expressed in all tissues, and its expression is not significantly affected in the ygl1 mutant. Interestingly, the mRNA expression of the cab1R gene encoding the Chl a/b-binding protein was severely suppressed in the ygl1 mutant. Moreover, the expression of some nuclear genes associated with Chl biosynthesis or chloroplast development was also affected in ygl1 seedlings. These results indicate that the expression of nuclear genes encoding various chloroplast proteins might be feedback regulated by the level of Chl or Chl precursors.     

Epidermal UV-shielding and photosystem II adjustment in wild type and chlorina f2 mutant of barley during exposure to increased PAR and UV radiation

The wild-type barley (WT; Hordeum vulgare L.) and its chlorophyll (Chl) b-less mutant chlorina f2 (clo f2) grown under shaded conditions in a greenhouse were transferred to outdoor conditions in early June with predominantly bright sunny days. During 6 days following transfer of plants we monitored the content of photosynthetic pigments, functional state of photosystem II (PSII) by means of Chl fluorescence induction kinetics and epidermal UV-shielding efficiency using Chl fluorescence imaging technique. Clo f2 mutant was more sensitive to exposure to an enhanced natural solar irradiance than WT barley. Nevertheless, clo f2 as well as WT were able to cope with stressful outdoor conditions, as was documented by the recovery of Chl a content and the maximal photochemical efficiency of PSII (F_V/F_M) after an initial decline. This was due to the immediate carotenoid-mediated photoprotection, reflected by strongly increased total carotenoids content and thermal energy dissipation localized within light-harvesting complexes of PSII (assessed by non-photochemical quenching of minimal fluorescence level). The positive acclimation response was further documented by an enhanced light-saturated electron transport rate through PSII (ETR). Based on the ratios of blue- to UV-excited Chl fluorescence we found that for both WT and clo f2 epidermal UV-shielding increased clearly after transfer to outdoor conditions and reached a saturation level after 3 days. In comparison with WT, clo f2 exhibited lower ability to induce UV-shielding. The kinetics of UV-shielding development during the outdoor treatment was different for the particular leaf regions. We suggest that this is related to the different age and developmental stage of the tissue along the leaf blade. The complementarity of carotenoid-mediated photoprotection and UV-shielding in acclimation of the assimilatory apparatus to increased visible and UV radiation is discussed.     

Photoinhibition and photoprotection in senescent leaves of field-grown wheat plants

Photosynthesis, photosystem II (PSII) photochemistry, photoinhibition and the xanthophyll cycle in the senescent flag leaves of wheat (Triticum aestivum L.) plants grown in the field were investigated. Compared to the non-senescent leaves, photosynthetic capacity was significantly reduced in senescent flag leaves. The light intensity at which photosynthesis was saturated also declined significantly. The light response curves of PSII photochemistry indicate that a down-regulation of PSII photochemistry occurred in senescent leaves in particular at high light. The maximal efficiency of PSII photochemistry in senescent flag leaves decreased slightly when measured at predawn but substantially at midday, suggesting that PSII function was largely maintained and photoinhibition occurred in senescent leaves when exposed to high light. At midday, PSII efficiency, photochemical quenching and the efficiency of excitation capture by open PSII centers decreased considerably, while non-photochemical quenching increased significantly. Moreover, compared with the values at early morning, a greater decrease in CO₂ assimilation rate was observed at midday in senescent leaves than in control leaves. The levels of antheraxanthin and zeaxanthin via the de-epoxidation of violaxanthin increased in senescent flag leaves from predawn to midday. An increase in the xanthophyll cycle pigments relative to chlorophyll was observed in senescent flag leaves. The results suggest that the xanthophyll cycle was activated in senescent leaves due to the decrease in CO₂ assimilation capacity and the light intensity for saturation of photosynthesis and that the enhanced formation of antheraxanthin and zeaxanthin at high light may play an important role in the dissipation of excess light energy and help to protect photosynthetic apparatus from photodamage. Our results suggest that the well-known function of the xanthophyll cycle to safely dissipate excess excitation energy is also important for maintaining photosynthetic function during leaf senescence.     

Gene cloning and functional analysis of yellow green leaf3 (ygl3) gene during the whole-plant growth stage in rice

Chlorophyll is an important photosynthetic pigment in the process of photosynthesis in plants and photosynthetic bacteria. Genes involved in chlorophyll biosynthesis in Arabidopsis and photosynthetic bacteria have been well documented. In rice, however, these genes have not been fully annotated. In this paper, a yellow-green leaf gene, yellow green leaf3 (ygl3) was cloned and analyzed. ygl3 encodes magnesium chelation ChlD (D) subunit, a key enzyme for chlorophyll synthesis, resulting in a yellow-green leaf phenotype in all growth stages in rice. Expression content of ygl3 is highest in the leaf blades, followed by the leaf sheaths, while there is virtually no expression of the gene in the stems and seeds. The sub-cellular structure and protein content of the photosynthetic system of the ygl3 mutant were revealed by transmission electron microscopy, BN-PAGE, and western blotting. The results show that the mutation of the ygl3 gene indirectly leads to a decrease in the protein content of the photosynthetic system and severely obstructs the formation of granum thylakoids.     

Young Leaf Chlorosis 1, a chloroplast-localized gene required for chlorophyll and lutein accumulation during early leaf development in rice

Chlorophyll (Chl) and lutein are the two most abundant and essential components in photosynthetic apparatus, and play critical roles in plant development. In this study, we characterized a rice mutant named young leaf chlorosis 1 (ylc1) from a ⁶⁰Co-irradiated population. Young leaves of the ylc1 mutant showed decreased levels of Chl and lutein compared to those of wild type, and transmission electron microscopy analysis revealed that the thylakoid lamellar structures were obviously loosely arranged. Whereas, the mutant turns green gradually and approaches normal green at the maximum tillering stage. The Young Leaf Chlorosis 1 (YLC1) gene was isolated via map-based cloning and identified to encode a protein of unknown function belonging to the DUF3353 superfamily. Complementation and RNA-interference tests confirmed the role of the YLC1 gene, which expressed in all tested rice tissues, especially in the leaves. Real-time PCR analyses showed that the expression levels of the genes associated with Chl biosynthesis and photosynthesis were affected in ylc1 mutant at different temperatures. In rice protoplasts, the YLC1 protein displayed a typical chloroplast location pattern. The N-terminal 50 amino acid residues were confirmed to be necessary and sufficient for chloroplast targeting. These data suggested that the YLC1 protein may be involved in Chl and lutein accumulation and chloroplast development at early leaf development in rice.     

Response of xanthophyll cycle and chloroplastic antioxidant enzymes to chilling stress in tomato over-expressing glycerol-3-phosphate acyltransferase gene

Over-expression of chloroplastic glycerol-3-phosphate acyltransferase gene (LeGPAT) increased unsaturated fatty acid contents in phosphatidylglycerol (PG) of thylakoid membrane in tomato. The effect of this increase on the xanthophyll cycle and chloroplast antioxidant enzymes was examined by comparing wild type (WT) tomato with the transgenic (TG) lines at chilling temperature (4 °C) under low irradiance (100 μmol m⁻² s⁻¹). Net photosynthetic rate and the maximal photochemical efficiency of photosystem (PS) 2 (F_v/F_m) in TG plants decreased more slowly during chilling stress and F_v/F_m recovered faster than that in WT plants under optimal conditions. The oxidizable P700 in both WT and TG plants decreased during chilling stress under low irradiance, but recovered faster in TG plants than in the WT ones. During chilling stress, non-photochemical quenching (NPQ) and the de-epoxidized ratio of xanthophyll cycle in WT plants were lower than those of TG tomatoes. The higher activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX) in TG plants resulted in the reduction of O₂ ^−· and H₂O₂ contents during chilling stress. Hence the increase in content of unsaturated fatty acids in PG by the over-expression of LeGPAT could alleviate photoinhibition of PS2 and PS1 by improving the de-epoxidized ratio of xanthophyll cycle and activities of SOD and APX in chloroplast.     

Photosystem 2 photochemistry and pigment composition of a yellow mutant of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) under different irradiances

A yellow leaf colouration mutant (named ycm) generated from rice T-DNA insertion lines was identified with less grana lamellae and low thylakoid membrane protein contents. At weak irradiance [50 μmol(photon) m⁻² s⁻¹], chlorophyll (Chl) contents of ycm were ≈20 % of those of WT and Chl a/b ratios were 3-fold that of wild type (WT). The leaf of ycm showed lower values in the actual photosystem 2 (PS2) efficiency (Φ_PS2), photochemical quenching (q_P), and the efficiency of excitation capture by open PS2 centres 1 (F_v′/F_m′) than those of WT, except no difference in the maximal efficiency of PS2 photochemistry (F_v/F_m). With progress in irradiance [100 and 200 μmol(photon) m⁻² s⁻¹], there was a change in the photosynthetic pigment stoichiometry. In ycm, the increase of total Chl contents and the decrease in Chl a/b ratio were observed. Φ_PS2, q_P, and F_v′/F_m′ of ycm increased gradually along with the increase of irradiance but still much less than in WT. The increase of xanthophyll ratio [(Z+A)/(V+A+Z)] associated with non-photochemical quenching (q_N) was found in ycm which suggested that ycm dissipated excess energy through the turnover of xanthophylls. No significant differences in pigment composition were observed in WT under various irradiances, except Chl a/b ratio that gradually decreased. Hence the ycm mutant developed much more tardily than WT, which was caused by low photon energy utilization independent of irradiance.     

Xanthophyll cycle induction by anaerobic conditions under low light in Chlamydomonas reinhardtii

The effect of anaerobiosis on the induction of the xanthophyll cycle was investigated in Chlamydomonas reinhardtii. The results showed that, anaerobiosis obtained by either sulfur starvation or by bubbling nitrogen in the culture grown in complete medium induced the xanthophyll cycle even when cultures were exposed to low light conditions. The zeaxanthin content reached 35 mmol mol^?1 Chl a, after 110 h in anaerobic sulfur-starved cultures, and 30 mmol mol^?1 Chl a within 24 h in sulfur replete cultures bubbled with nitrogen. Both starved and non-starved cultures grown under aerobic conditions, did not exhibit any sizeable increase in the zeaxanthin content. Chlorophyll fluorescence measurements revealed a decrease in the maximum photochemical quantum yield of PSII (F_v/F_m) by more than 50 %. The chlorophyll fluorescence kinetics (OJIP) analysis showed a strong rise at the J-step indicating a strong reduction of Q_A. Our findings demonstrated that anaerobiosis in low light exposed cultures induced the xanthophyll cycle through a strong increase of the level of plastoquinone pool reduction, which was associated to the formation of a trans-thylakoid membranes proton gradient, while in dark anaerobic cultures, no appreciable induction of xanthophyll cycle could be observed, despite the sizeable increase in non–photochemical quenching.     

The role of the xanthophyll cycle and of lutein in photoprotection of photosystem II

Photoprotection of photosystem II (PSII) is essential to avoid the light-induced damage of the photosynthetic apparatus due to the formation of reactive oxygen species (=photo-oxidative stress) under excess light. Carotenoids are known to play a crucial role in these processes based on their property to deactivate triplet chlorophyll (3Chl*) and singlet oxygen (1O?*). Xanthophylls are further assumed to be involved either directly or indirectly in the non-photochemical quenching (NPQ) of excess light energy in the antenna of PSII. This review gives an overview on recent progress in the understanding of the photoprotective role of the xanthophylls zeaxanthin (which is formed in the light in the so-called xanthophyll cycle) and lutein with emphasis on the NPQ processes associated with PSII of higher plants. The current knowledge supports the view that the photoprotective role of Lut is predominantly restricted to its function in the deactivation of 3Chl*, while zeaxanthin is the major player in the deactivation of excited singlet Chl (1Chl*) and thus in NPQ (non-photochemical quenching). Additionally, zeaxanthin serves important functions as an antioxidant in the lipid phase of the membrane and is likely to act as a key component in the memory of the chloroplast with respect to preceding photo-oxidative stress. This article is part of a Special Issue entitled: Photosystem II.     

Adaptation of the Photosynthetic Apparatus in Maize Leaves as a Result of Nitrogen Limitation : Relationships between Electron Transport and Carbon Assimilation

In maize (Zea mays L., cv Contessa), nitrogen (NO₃⁻) limitation resulted in a reduction in shoot growth and photosynthetic capacity and in an increase in the leaf zeaxanthin contents. Nitrogen deficiency had only a small effect on the quantum yield of CO₂ assimilation but a large effect on the light-saturated rate of photosynthesis. Linear relationships persisted between the quantum yield of CO₂ assimilation and that of photosystem II photochemistry in all circumstances. At high irradiances, large differences in photochemical quenching and nonphotochemical quenching of Chl a fluorescence as well as the ratio of variable to maximal fluorescence (Fv/Fm) were apparent between nitrogen-deficient plants and nitrogen-replete controls, whereas at low irradiances these parameters were comparable in all plants. Light intensity-dependent increases in nonphotochemical quenching were greatest in nitrogen-deficient plants as were the decreases in Fv/Fm ratio. In nitrogen-deficient plants, photochemical quenching decreased with increasing irradiance but remained higher than in controls at high irradiances. Thermal dissipative processes were enhanced as a result of nitrogen deficiency (nonphotochemical quenching was elevated and Fv/Fm was lowered) allowing PSII to remain relatively oxidised even when carbon metabolism was limited via nitrogen limitation.     

Photosynthetic responses of Coffea arabica leaves to a short-term high light exposure in relation to N availability

It is known that the coffee (Coffea arabica L.) plant which is originally from shade habitats would have a limited ability to grow under full sun. Previous work has shown that nitrogen fertilisation can reduce the leaf damage when the plants are exposed to high light intensities during several days. In the present work we aimed to study the effects of the high irradiance during the first hours and evaluate the positive contribution of nitrogen fertilisation in the case of short-term exposure to strong light. Young plants (1.5–2 years old) grown in 1.5 kg of a mixed soil were supplemented with a nutrient solution containing 15 mM nitrogen in the form of NH₄NO₃, every 7 days (2N treatment), 15 days (1N treatment) and 45 days (0N treatment). Top mature leaves were exposed to a photosynthetic photon flux density of 1 500 μmol m^?2 s^?1 for a maximal period of 8 h, and changes in photosynthesis and pigment composition were monitored along the period of high light exposure. Photosynthetic capacity, leaf conductance to water vapour, electron transport capacity and maximum carboxylation activity, as well as some leaf fluorescence parameters (minimal fluorescence, photochemical efficiency of PSII and quantum yield of photosynthetic electron transport) were reduced by the stress, with a generally stronger impact observed in the 0N plants. The photochemical quenching was affected only in the 0N plants, while the non-photochemical quenching increased in 2N plants but decreased in the 0N ones. The results showed that 2N plants presented a better initial status of the photosynthetic parameters and of the content of photoprotective pigments. Those plants showed ability to trigger some protective mechanisms, as observed by the tendency to increase the xanthophyll pool content, specially in zeaxanthin and in non-photochemical quenching. Also, protein content presented a tendency to increase after 1.5 h, which was maintained until the end of the high light period. We conclude that nitrogen availability is a key factor in the acclimation process to high light.     

Photosynthesis and chlorophyllafluorescence during flag leaf senescence of field-grown wheat plants

The characteristics of photosynthetic gas exchange, chlorophyll a fluorescence, and xanthophyll cycle pigments during flag leaf senescence of field-grown wheat plants were investigated. With senescence progressing, the light-saturated net CO₂ assimilation rate expressed either on a basis of leaf area or chlorophyll decreased significantly. The apparent quantum yield of net photosynthesis decreased when expressed on a leaf area basis but increased when expressed on a chlorophyll basis. The maximal efficiency of PSII photochemistry decreased very little while actual PSII efficiency, photochemical quenching, and the efficiency of excitation capture by open PSII centers decreased considerably. At the same time, non-photochemical quenching increased significantly. A substantial decrease in the contents of violaxanthin and zeaxanthin, but a slight decrease in the content of antheraxanthin were observed. However, the de-epoxidation status of the xanthophyll cycle was positively correlated with progressive senescence. This increase was due mainly to a smaller decrease in zeaxanthin than in violaxanthin. Our results suggest that PSII apparatus remained functional, but a down-regulation of PSII occurred under the steady state of photosynthesis in senescent flag leaves. Such a down-regulation was associated with the closure of PSII centers and an enhanced xanthophyll cycle-related thermal dissipation in the PSII antennae.     

The aba Mutant of Arabidopsis thaliana (L.) Heynh. Has Reduced Chlorophyll Fluorescence Yields and Reduced Thylakoid Stacking

It has been shown that the aba mutant of Arabidopsis thaliana (L.) Heynh. is impaired in epoxy-carotenoid biosynthesis and accumulates the epoxy-carotenoid precursor, zeaxanthin (C.D. Rock, J.A.D. Zeevaart [1991] Proc Natl Acad Sci USA 88: 7496-7499). In addition to providing conclusive evidence for the indirect pathway of abscisic acid biosynthesis from epoxy-carotenoids, the aba mutation offers a powerful means to study the function of xanthophylls (oxygenated carotenoids) in photosynthesis. We measured in vivo the chlorophyll (Chl) fluorescence parameters F_o (initial), F_m (maximum), F_v (variable = F_m − F_o), and t_½ (half-rise time of fluorescence induction) of wild-type (WT) and three allelic aba mutants. The mutant genotypes had significantly lower F_o and F_m values relative to those of WT. The F_v/F_m ratio and t_½, which are parameters affected by photochemical efficiency, photosystem II (PSII), and plastoquinone pool sizes, were similar in the aba alleles and WT. Because the aba genotypes accumulate high levels of zeaxanthin, which is involved in nonphotochemical quenching of Chl fluorescence, we propose that the reduced fluorescence yields in the aba genotypes are a consequence of the accumulated zeaxanthin. Measurement of PSII oxygen evolution rates in isolated thylakoid membranes of WT and aba-4 confirmed that quantum efficiency was not altered in aba-4 but indicated that the mutant had reduced PSII activity in vitro. Electron microscopy revealed an abnormal chloroplast ultrastructure in the aba plants: the mutants had significantly fewer thylakoid lamellae per granum stack but significantly more grana per chloroplast, as well as more chloroplasts per cell than WT. Immunoblot analysis established that aba-4 had normal levels of the Chl a/b-binding core polypeptide of PSII (CP29) and the PSII light-harvesting Chl a/b-binding complex. These results provide evidence for the role of zeaxanthin in nonphotochemical fluorescence quenching and suggest involvement of epoxy-carotenoids and/or zeaxanthin in thylakoid stacking and PSII activity.     

Monitoring cashew seedlings during interactions with the fungus <Emphasis Type="Italic">Lasiodiplodia theobromae</Emphasis> using chlorophyll fluorescence imaging

The chlorophyll (Chl) fluorescence imaging technique was applied to cashew seedlings inoculated with the fungus Lasiodiplodia theobromae to assess any disturbances in the photosynthetic apparatus of the plants before the onset of visual symptoms. Two-month-old cashew plants were inoculated with mycelium of L. theobromae isolate Lt19 or Lt32. Dark-adapted and light-acclimated whole plants or previously labelled, single, mature leaf from each plant were evaluated weekly for Chl fluorescence parameters. From 21 to 28 days, inoculation with both isolates resulted in the significantly lower maximal photochemical quantum yield of PSII (F_v/F_m) than those for control samples, decreasing from values of 0.78 to 0.62. In contrast, the time response of the measured fluorescence transient curve from dark-acclimated plants increased in both whole plants and single mature leaves in inoculated plants compared with controls. The F_v/F_m images clearly exhibited photosynthetic perturbations 14 days after inoculation before any visual symptoms appeared. Additionally, decays in the effective quantum yield of PSII photochemistry and photochemical quenching coefficient were also observed over time. However, nonphotochemical quenching increased during the evaluation period. We conclude that F_v/F_m images are the effective way of detecting early metabolic perturbations in the photosynthetic apparatus of cashew seedlings caused by gummosis in both whole plants and single leaves and could be potentially employed in larger-scale screening systems.     

Ecophysiological responses of Caragana korshinskii Kom. under extreme drought stress: Leaf abscission and stem survives

Caragana korshinskii Kom. is a perennial xerophytic shrub, well known for its ability to resist drought. In order to study ecophysiological responses of C. korshinskii under extreme drought stress and subsequent rehydration, diurnal patterns of gas exchange and chlorophyll (Chl) fluorescence parameters of photosystem II as well as Chl content were analyzed. Plant responses to extreme drought included (1) leaf abscission and using stem for photosynthesis, (2) improved instantaneous water-use efficiency, (3) decreased photosynthetic rate and partly closed stomata owing to leaf abscission and low water status, (4) decreased maximum photochemical efficiency of photosystem II (PSII) (variable to maximum fluorescence ratio, F_v/F_m), quantum efficiency of noncyclic electron transport of PSII, and Chl a and Chl b. Four days after rehydration, new leaves budded from stems. In the rewatered plants, the chloroplast function was restored, the gas exchange and Chl fluorescence returned to a similar level as control plant. The above result indicated that maintaining an active stem system after leaf abscission during extreme drought stress may be the foundation which engenders these mechanisms rapid regrowth for C. korshinskii in arid environment.