Chlorophyll Bleaching Research Articles

Preharvest Application of Sodium Nitroprusside Alleviates Yellowing of Chinese Flowering Cabbage via Modulating Chlorophyll Metabolism and Suppressing ROS Accumulation.

Chinese flowering cabbage is prone to senescence and yellowing after harvest, leading to a huge postharvest loss. Nitric oxide (NO) is a multifunctional plant growth regulator, but the effect of preharvest application of NO on the storage quality of Chinese flowering cabbage remains unclear. Preharvest application of 50 mg L-1 sodium nitroprusside (SNP, a NO donor) to the roots obviously reduced leaf yellowing in Chinese flowering cabbage during storage. Proteomic analysis reveals 198 differentially expressed proteins (DEPs) in SNP-treated plants compared to the control. The main DEPs were significantly enriched in chlorophyll metabolisms, phenylpropanoid synthesis, and antioxidant pathways. SNP treatment enhanced chlorophyll biosynthesis and suppressed chlorophyll-degradation-related proteins and genes. It also modulated flavonoid-biosynthesis-related genes, and 21 significantly regulated flavonoids were identified in SNP-treated plants. The enhanced antioxidant capacity in SNP-treated plants was able to decrease chlorophyll catabolism by inhibiting peroxidase-mediated chlorophyll bleaching. Collectively, preharvest SNP treatment modulated chlorophyll metabolism and preserved chlorophyll content in leaves during storage. Moreover, SNP treatment enhanced flavonoid synthesis, suppressed reactive oxygen species accumulation, and delayed the senescence process, thereby maintaining leaf greening in Chinese flowering cabbage. These findings highlight the role of exogenous NO in alleviating yellowing of leafy vegetables.

Temporal variation of 2-MIB and geosmin production by Pseudanabaena galeata and Phormidium ambiguum exposed to high-intensity light.

This study demonstrated the temporal variation of 2-methylisoborneol (2-MIB) and geosmin (GSM) production of two filamentous cyanobacteria species Pseudanabaena galeata (NIES-512; planktonic) and Phormidium ambiguum (NIES-2119; benthic) exposed to high light intensity (950-1000 μmol m-2 s-1 photosynthetically active radiation). The production of 2-MIB and GSM was quantified together with oxidative stress, chlorophyll content, and cellular protein content. The relative chlorophyll bleaching and cell degradations were compared through microscopic images. The 2-MIB production of P.galeata increased by over 42 ± 17% on the second day of exposure and remained leveled through the exposure period. P.ambiguum showed a continuous increase of 2-MIB until the 10th day, recording a 95 ± 4% increment. The GSM production was elevated until the fourth day of exposure by 46 ± 10% for P.galeata and by 74 ± 21% on the second day for P.ambiguum and reduced with prolonged exposure for both species. The chlorophyll content of P.galeata was reduced by 62 ± 7% on the second day, and that of P.ambiguum was reduced by 52 ± 9% on the fourth day and remained low. Protein and H2 O2 contents of both species were changed inconsistently. Exposure to high-intensity light can photobleach and deteriorate cells of both species, but elevations in odorous compounds can be expected.

HyPRP1, A Tomato Multipotent Regulator, Negatively Regulates Tomato Resistance to Sulfur Dioxide Toxicity and Can Also Reduce Abiotic Stress Tolerance of Escherichia coli and Tobacco

Abiotic stresses have led to an extensive decline in global crop production and quality. As one of the abiotic stress factors, sulfur dioxide (SO2) causes severe oxidative damage to plant tissues. Based on our previous study, a tomato hybrid-proline-rich protein 1 (HyPRP1) was found to be involved in abiotic stress and SO2 metabolism, though the gene functions remained largely unknown. In this study, the function analysis of the HyPRP1 gene was extended, and DNA methylation analysis, subcellular localization, and cis-element analysis were performed to investigate the features of this gene. The DNA methylation analysis implied that the HyPRP1 gene was hypermethylated and the methylation density in the leaf differed from that in the flower and fruit. Subcellular localization analysis identified HyPRP1 localized in the cytoplasm and plasma membrane in vivo. The E. coli cells harboring SlHyPRP1 showed reduced salt and drought resistance. In tomato, when SO2 toxicity occurred, the HyPRP1 RNAi knockdown lines accumulated more sulfates and less hydrogen peroxide (H2O2) and showed minimal leaf necrosis and chlorophyll bleaching. In tobacco, the overexpression of HyPRP1 reduced tolerance against salt stresses exerted by NaCl. We conclude that the heterologous expression of tomato HyPRP1 in E. coli and tobacco reduces abiotic stress tolerance and negatively regulates the resistance to sulfur dioxide toxicity by scavenging H2O2 and sulfite in tomato.

Molecular Characterization of Pseudomonas syringae pv. coriandricola and Biochemical Changes Attributable to the Pathological Response on Its Hosts Carrot, Parsley, and Parsnip.

Bacterial leaf spot caused by the plant pathogenic bacterium Pseudomonas syringae pv. coriandricola (Psc) was observed on carrot, parsnip, and parsley grown on a vegetable farm in the Vojvodina Province of Serbia. Nonfluorescent bacterial colonies were isolated from diseased leaves and characterized using different molecular techniques. Repetitive element PCR fingerprinting with five oligonucleotide primers (BOX, ERIC, GTG5, REP, and SERE) and the randomly amplified polymorphic DNA-PCR with the M13 primer revealed identical fingerprint patterns for all tested strains. Multilocus sequence analysis of four housekeeping genes (gapA, gltA, gyrB, and rpoD) showed a high degree (99.8 to 100%) of homology with sequences of Psc strains deposited in the Plant-Associated Microbes Database and NCBI database. The tested strains caused bacterial leaf spot symptoms on all three host plants. Host-strain specificity was not found in cross-pathogenicity tests, but the plant response (peroxidase induction and chlorophyll bleaching) was more pronounced in carrot and parsley than in parsnip.

Species-species interactions modulate copper toxicity under different visible light conditions

Combination of biotic and abiotic factors influences the effects of naturally occurring or anthropogenic chemicals on photosynthetic microorganisms in the aquatic environment. Nonetheless, the combined effects of physical stressors and species-species interaction on chemicals’ toxicity are still poorly understood. The present study examines the responses of the green alga Chlamydomonas reinhardtii and the cyanobacterium Synechocystis sp. alone and in mixtures to copper exposure under increasing visible light intensities. Cell growth, chlorophyll bleaching, oxidative stress and membrane permeability were determined by flow cytometry in both mono- and multi-species tests. The results revealed that species-species interactions influenced copper toxicity under different light regimes at 4 h and 48 h - exposure. For a given light condition, monocultures of Synechocystis sp. were more sensitive to copper than those of C. reinhardtii. In long-term incubation C. reinhardtii sensitivity to copper diminished in presence of Synechocystis sp. under low-intensity light, however it was enhanced under high-intensity light. The present results revealed the complex interplay between visible light intensity variations, species-species interaction and copper effects to phytoplankton in long- term exposure.

AtPep3 is a hormone-like peptide that plays a role in the salinity stress tolerance of plants

Peptides encoded by small coding genes play an important role in plant development, acting in a similar manner as phytohormones. Few hormone-like peptides, however, have been shown to play a role in abiotic stress tolerance. In the current study, 17 Arabidopsis genes coding for small peptides were found to be up-regulated in response to salinity stress. To identify peptides leading salinity stress tolerance, we generated transgenic Arabidopsis plants overexpressing these small coding genes and assessed survivability and root growth under salinity stress conditions. Results indicated that 4 of the 17 overexpressed genes increased salinity stress tolerance. Further studies focused on AtPROPEP3, which was the most highly up-regulated gene under salinity stress. Treatment of plants with synthetic peptides encoded by AtPROPEP3 revealed that a C-terminal peptide fragment (AtPep3) inhibited the salt-induced bleaching of chlorophyll in seedlings. Conversely, knockdown AtPROPEP3 transgenic plants exhibited a hypersensitive phenotype under salinity stress, which was complemented by the AtPep3 peptide. This functional AtPep3 peptide region overlaps with an AtPep3 elicitor peptide that is related to the immune response of plants. Functional analyses with a receptor mutant of AtPep3 revealed that AtPep3 was recognized by the PEPR1 receptor and that it functions to increase salinity stress tolerance in plants. Collectively, these data indicate that AtPep3 plays a significant role in both salinity stress tolerance and immune response in Arabidopsis.

Bleaching of biofilm-forming algae induced by UV-C treatment: a preliminary study on chlorophyll degradation and its optimization for an application on cultural heritage.

Green microalgae colonizing stone surfaces represent a major problem for the conservation of heritage monuments, since they lead to biodegradation and aesthetic issues. Previous studies in La Glacière show cave (France) have demonstrated that UV-C may have a strong effect on microalgae, thus leading to chlorophyll bleaching, which was increased when biofilms were maintained under VIS-light condition unlike to those maintained in the dark. To understand the physiological mechanisms underlying this response and in order to optimize in situ treatment, 30kJm-2 UV-C exposure times were applied to Chlorophyta Chlorella sp. and chlorophyll degradation kinetics were then monitored. UV-C irradiation was enough to inhibit photosynthesis and to directly kill all algal cells. Results also showed that chlorophyll a was degraded faster than chlorophyll b and that 14h were necessary for complete degradation of all the present chlorophyll. In addition, our results highlighted the importance of visible light exposition after UV-C treatment which leading to chlorophyll bleaching. Irradiated algae cultivated in the dark were still green 5days after treatment while cultivated samples in the light lost their green color after 14h. An efficient UV-C treatment applicable to show caves and other heritage monuments was proposed.

Effects of UV-C radiation on Chlorella vulgaris, a biofilm-forming alga

Photosynthetic biofilms proliferating on heritage monuments represent a major threat for curators leading to biodegradation and esthetic issues. Previous studies demonstrated that UV-C, used as a tool for biofilm eradication, is a promising avenue to combat microbial proliferation. In this study, this environmentally friendly method was tested on biofilm-forming Chlorella vulgaris suspension. Algal physiological response to UV-C was then assessed. Results showed that > 10 kJ m−2 UV-C exposure was enough to directly kill cells whereas low UV-C exposure reduced quantum yield of photosystem II and inhibited both respiration and photosynthesis. Clear relationships between UV-C exposure times and physiological responses were found. In addition, the use of VIS-light after UV-C treatment enhances chlorophyll bleaching. Our findings contribute to a better understanding of the physiological responses of Chlorella vulgaris to UV-C radiation allowing thus an optimization of the UV-C treatment reported in our previous studies.

Thermo-Induced Maintenance of Photo-oxidoreductases Underlies Plant Autotrophic Development

Chlorophyll biosynthesis enables autotrophic development of developing seedlings. Upon light exposure, the chlorophyll precursor protochlorophyllide produces reactive oxygen species (ROS). Developingseedlings acquire photosynthetic competence through the action of protochlorophyllide oxidoreductases (PORs) that convert protochlorophyllide to chlorophyllide, reducing ROS production that would otherwise induce cellular damage and chlorophyll bleaching. Here, we show that FCA mediates the thermostabilization of PORs to trigger the conversion of protochlorophyllide to chlorophyllide in developing seedlings. FCA also facilitates the thermal induction of POR genes through histone acetylation that promotes the accessibility of RNA polymerasesto the gene promoters. The combined action of FCA maintains PORs at warm temperatures, shifting the chlorophyll-ROS balance toward autotrophic development. We propose that the FCA-mediated thermal adaptation of autotrophic development allows developing seedlings to cope with the heat-absorbing soil surface layer under natural conditions. The thermal adaptive mechanism would provide apotential basis for studying crop performance at warm temperatures.

Improvement of biomass accumulation of potato plants by transformation of cyanobacterial photorespiratory glycolate catabolism pathway genes

Transgenic potato (Solanum tuberosum L. cv. Desiree) plants expressing components of a novel cyanobacterial photorespiratory glycolate catabolism pathway were developed. Transgenic plant expressing glcD1 (glycolate dehydrogenase I) gene was referred to as synGDH and transgenic plants expressing gcl (glyoxylate carboligase) and tsr (tartronic semialdehyde reductase) genes simultaneously were designated as synGT. Both synGDH and synGT plants showed stable gene transformation, integration and expression. Enhanced glyoxylate contents in synGDH plants were detected as compared to synGT and non-transgenic (NT) plants. Phenotypic evaluation revealed that synGDH plants accumulated 11 % higher dry weight, while, tuber weight was 38 and 16 % higher than NT and synGT, respectively. Upon challenging the plants in high temperature and high light conditions synGDH plants maintained higher Fv/Fm and showed less bleaching of chlorophyll as compared to synGT and NT plants. These results indicate that genetic transformation of complete pathway in one plant holds promising outcomes in terms of biomass accumulation to meet future needs for food and energy.

Symbiodinium sp. cells produce light-induced intra- and extracellular singlet oxygen, which mediates photodamage of the photosynthetic apparatus and has the potential to interact with the animal host in coral symbiosis.

Coral bleaching is an important environmental phenomenon, whose mechanism has not yet been clarified. The involvement of reactive oxygen species (ROS) has been implicated, but direct evidence of what species are involved, their location and their mechanisms of production remains unknown. Histidine-mediated chemical trapping and singlet oxygen sensor green (SOSG) were used to detect intra- and extracellular singlet oxygen ((1) O2 ) in Symbiodinium cultures. Inhibition of the Calvin-Benson cycle by thermal stress or high light promotes intracellular (1) O2 formation. Histidine addition, which decreases the amount of intracellular (1) O2 , provides partial protection against photosystem II photoinactivation and chlorophyll (Chl) bleaching. (1) O2 production also occurs in cell-free medium of Symbiodinium cultures, an effect that is enhanced under heat and light stress and can be attributed to the excretion of (1) O2 -sensitizing metabolites from the cells. Confocal microscopy imaging using SOSG showed most extracellular (1) O2 around the cell surface, but it is also produced across the medium distant from the cells. We demonstrate, for the first time, both intra- and extracellular (1) O2 production in Symbiodinium cultures. Intracellular (1) O2 is associated with photosystem II photodamage and pigment bleaching, whereas extracellular (1) O2 has the potential to mediate the breakdown of symbiotic interaction between zooxanthellae and their animal host during coral bleaching.

Ambient UV-B radiation inhibits the growth and physiology of Brassica napus L. on the Qinghai-Tibetan plateau

Increased amounts of ultraviolet-B (UV-B) radiation reaching the Earth's surface through stratospheric ozone depletion are expected to have a negative effect on plant responses. However, the reliability of extrapolating indoor experiments to infer plant responses under field conditions has been questioned. Here, we report the growth and physiological responses of Brassica napus L. crops grown on the Qinghai-Tibetan plateau to different levels of ambient UV-B radiation (100%, 70%, and 25%). Here, we aimed to obtain a realistic evaluation of the effect of high UV-B radiation on B. napus L. crops in this region. We used three experimental groups: control (ambient UV-B radiation), T1 (25% exclusion of solar UV-B), and T2 (70% exclusion of solar UV-B). Compared to the control, exclusion of solar UV-B radiation enhanced specific leaf weight (SLW) and caused plant height increased with a significant increase in biomass. Ambient UV-B radiation caused the UV-B absorbing compounds of the leaves to increase, while chlorophyll a, b, and (a+b) content decreased. No significant differences in carotenoid content were detected among the three groups. Compared to the control, exclusion of solar UV-B radiation reduced antioxidant enzyme activity. Moreover, the results showed that exposure to UV-B radiation caused B. napus to (i) increase UV-B absorbing compounds to reduce the transmittance of UV photons through the leaf tissue, (ii) enhance antioxidant enzyme activity to scavenge reactive oxygen species (ROS), and (iii) increase carotenoids to prevent oxidative damage. However, the bleaching of chlorophyll a and damage to the photosynthetic apparatus by solar UV-B radiation caused a reduction in the photosynthetic rate.

Cellular and molecular damage caused by high UV-C irradiation of the cave-harvested green alga Chlorella minutissima: Implications for cave management

Algal biofilms in caves visited by tourists are a significant threat to karstic formations. They cause a dirty greenish appearance on cave formations and provoke biophysical and biochemical erosion. Seeking an alternative to the use of chemicals, we investigated the effects of high doses of UV-C irradiation on the green algae Chlorella minutissima-dominating biofilms in the Moidons Cave (Jura, France). The algae were harvested and subjected to laboratory subculturing under conditions similar to the conditions in the cave (low temperatures and a short photoperiod with weak light intensities). After one or two doses of UV-C of 150 or 300 kJ m−2, the samples were incubated for 21 days, and the physiological parameters (pigment concentrations and photosynthetic activity) were monitored every seven days. Hydrogen peroxide (H2O2) and malondialdehyde (MDA) concentrations were also measured to detect oxidative stress, and analyses of DNA integrity were performed. The results showed that all UV-C treatments caused chlorophyll bleaching with completely inhibited metabolic activity. In addition, high UV-C doses caused oxidative stress and programmed cell death. Irradiation was tested on a natural biofilm that proliferated in the Moidons Cave. Colorimetric measurements of the treated biofilm indicated that UV-C treatment caused significant chlorophyll bleaching.

Phytotoxic effects of selected N-benzyl-benzoylhydroxamic acid metallo-oxygenase inhibitors: investigation into mechanism of action

Treatment of Arabidopsis thaliana with 100 μM hydroxamic acids F1 and F2, found previously to inhibit carotenoid cleavage dioxygenase enzyme CCD1, was found to cause chlorophyll bleaching and phytotoxicity. A further set of hydroxamic acid analogues was synthesised, and these compounds were found to be phytotoxic towards A. thaliana at 16–400 μM, and to show some phytoxicity towards broad-leaved weeds C. album and S. media at 100 μM. Compound F1 was found to inhibit p-hydroxy-phenylpyruvate dioxygenase (HPPD), a known herbicide target (IC50 30 μM), but compounds F5 and F8 showed no inhibition of HPPD, despite F8 showing higher levels of phytotoxicity. Plants grown in the presence of F1 or F5 that were treated with 50 μM homogentisic acid showed partial recovery of growth, indicating some inhibition of HPPD in planta. These are the first hydroxamic acid inhibitors reported for HPPD, but the results indicate that inhibition of HPPD is only partly responsible for the observed phytotoxicity, and that another unknown metalloenzyme is also targeted by these compounds.

Sulfite reductase protects plants against sulfite toxicity.

Plant sulfite reductase (SiR; Enzyme Commission 1.8.7.1) catalyzes the reduction of sulfite to sulfide in the reductive sulfate assimilation pathway. Comparison of SiR expression in tomato (Solanum lycopersicum 'Rheinlands Ruhm') and Arabidopsis (Arabidopsis thaliana) plants revealed that SiR is expressed in a different tissue-dependent manner that likely reflects dissimilarity in sulfur metabolism between the plant species. Using Arabidopsis and tomato SiR mutants with modified SiR expression, we show here that resistance to ectopically applied sulfur dioxide/sulfite is a function of SiR expression levels and that plants with reduced SiR expression exhibit higher sensitivity than the wild type, as manifested in pronounced leaf necrosis and chlorophyll bleaching. The sulfite-sensitive mutants accumulate applied sulfite and show a decline in glutathione levels. In contrast, mutants that overexpress SiR are more tolerant to sulfite toxicity, exhibiting little or no damage. Resistance to high sulfite application is manifested by fast sulfite disappearance and an increase in glutathione levels. The notion that SiR plays a role in the protection of plants against sulfite is supported by the rapid up-regulation of SiR transcript and activity within 30 min of sulfite injection into Arabidopsis and tomato leaves. Peroxisomal sulfite oxidase transcripts and activity levels are likewise promoted by sulfite application as compared with water injection controls. These results indicate that, in addition to participating in the sulfate assimilation reductive pathway, SiR also plays a role in protecting leaves against the toxicity of sulfite accumulation.

Improved Cold and Drought Tolerance of Doubled Haploid Maize Plants Selected for Resistance to Prooxidant tert-Butyl Hydroperoxide

To improve the abiotic stress tolerance of maize (Zea mays L.), doubled haploid (DH) plants were produced by in vitro selection of microspores exposed to tert-butyl hydroperoxide (t-BuOOH) as a powerful prooxidant This study investigated the tolerance of the progenies of t-BuOOH-selected DH lines to oxidative stress, cold and drought in controlled environment pot experiments by analyses of photosynthetic electron transport and CO2 assimilation processes, chlorophyll bleaching and lipid peroxidation of leaves. Our results demonstrated that the t-BuOOH-selected DH plants exhibited enhanced tolerance not only to oxidative stress-induced by t-BuOOH but also to cold and drought stresses. In addition, they showed elevated activities of antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, catalase, glutathione reductase and glutathione S-transferase when compared with the DH lines derived from microspores that were not exposed to t-BuOOH and to the original hybrid plants. The results suggest that the simultaneous up-regulation of several antioxidant enzymes may contribute to the oxidative and cold stress tolerance of the t-BuOOH-selected DH lines, and that the in vitro microspore selection represents a potential way to improve abiotic stress tolerance in maize.

Photoinduced changes in photosystem II pigments

The photosynthetic apparatus in higher plants performs two seemingly opposing tasks: efficient harvest of sunlight, but also rapid and harmless dissipation of excess light energy as heat to avoid deleterious photodamage. In order to study this process in pigment-protein supercomplexes of photosystem II (PSII), 77 K fluorescence and room temperature resonance Raman (RR) spectroscopy were applied to investigate the changes in structure and spectral properties of the pigments in spinach PSII membranes. The high-light treatment results in a strong quenching of the fluorescence (being largest when the excitation is absorbed by carotenoids) and a red-shift of the main maximum. Decomposition of the fluorescence spectra into four bands revealed intensive quenching of F685 and F695 bands, possible bleaching of chlorophyll a, enhanced extent of light harvesting complexes (LHCII) aggregation and increased energy transfer to aggregated LHCII. The analysis of RR spectra revealed the predominant contribution of ß-carotene (ß-Car) upon 457.8 and 488 nm excitations and lutein (Lut) at 514.5 nm. During prolonged exposure to strong light no significant bleaching of ß-Car and weak photobleaching of Lut is observed. The results will contribute to the efforts to produce more efficient and robust solar cells when exposed to fluctuations in light intensity.

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Purification and characterization of a stable oxygen-evolving Photosystem II complex from a marine centric diatom, Chaetoceros gracilis

Oxygen-evolving Photosystem II particles (crude PSII) retaining a high oxygen-evolving activity have been prepared from a marine centric diatom, Chaetoceros gracilis (Nagao et al., 2007). The crude PSII, however, contained a large amount of fucoxanthin chlorophyll a/c-binding proteins (FCP). In this study, a purified PSII complex which was deprived of major components of FCP was isolated by one step of anion exchange chromatography from the crude PSII treated with Triton X-100. The purified PSII was still associated with the five extrinsic proteins of PsbO, PsbQ', PsbV, Psb31 and PsbU, and showed a high oxygen-evolving activity of 2135 μmol O2 (mg Chl a)−1 h−1 in the presence of phenyl-p-benzoquinone which was virtually independent of the addition of CaCl2. This activity is more than 2.5-fold higher than the activity of the crude PSII. The activity was completely inhibited by 3-(3,4)-dichlorophenyl-(1,1)-dimethylurea (DCMU). The purified PSII contained 42 molecules of Chl a, 2 molecules of diadinoxanthin and 2 molecules of Chl c on the basis of two molecules of pheophytin a, and showed typical absorption and fluorescence spectra similar to those of purified PSIIs from the other organisms. In this study, we also found that the crude PSII was significantly labile, as a significant inactivation of oxygen evolution, chlorophyll bleaching and degradation of PSII subunits were observed during incubation at 25 °C in the dark. In contrast, these inactivation, bleaching and degradation were scarcely detected in the purified PSII. Thus, we succeeded for the first time in preparation of a stable PSII from diatom cells.

Photocatalytic inhibition of cyanobacterial growth using silver-doped TiO2 under UV-C light

Capacity of the silver-doped TiO2 under UV-C light to restrain cyanobacterial growth was explored with Anabaena sp. PCC 7120 and Microcystis aeruginosa as test species. The survival, chlorophyll bleaching, photosynthetic activity, DNA breakages, antioxidant enzyme activities, lipid peroxidation, and cellular morphologic structure of test cyanobacteria were analyzed. The results indicate that the test cyanobacteria with UV-C photocatalysis by silver-doped TiO2 sufferd from effects of reactive oxygen species, which promote the damage of the cell wall and the peroxidation of cell membranes, and subsequently, aggravate the losses of cell organelle and viability. The results suggest that UV-C photocatalysis by the silver ions doped TiO2 could be a promising method to prevent fast and excessive growth of cyanobacteria in eutrophic water sources.