This study summarizes the response of cyanobacterium Spirulina subsalsa HKAR-19 under simulated light conditions of photosynthetically active radiation (PAR), PAR+UV-A (PA), and PAR+UV-A+UV-B (PAB). Exposure to UV radiation caused a significant (P < 0.05) decrease in chlorophyll a, phycocyanin, and total protein. In contrast, total carotene content increased significantly (P < 0.05) under PA and PAB with increasing irradiation time. The photosynthetic efficiency of photosystem II also decreased significantly in PA and PAB radiation. We have also recorded a decrease in the fluorescence emission intensity of phycocyanin under PA and PAB exposure. The phycocyanin fluorescence shifted towards shorter wavelengths (blue-shift) after 72 h of PA and PAB exposure. Intracellular reactive oxygen species (ROS) levels increased significantly in PA and PAB. Fluorescence microscopic images showed an increase in green fluorescence, indicating ROS generation in UV radiation. We have also quantified ROS generation using green and red fluorescence ratio represented as G/R ratio. A 2-6-fold increase in antioxidative enzymes activity was observed to overcome the damaging effects caused by UV stress as compared to untreated control cultures. The lipid peroxidation was assessed in terms of malondialdehyde content which increases significantly (P < 0.05) as the duration of exposure increases. These results suggest that a combined effect of PAR, UV-A, and UV-B was more deleterious than an individual one.
- MeSH
- antioxidancia * metabolismus MeSH
- chlorofyl a metabolismus MeSH
- chlorofyl * metabolismus MeSH
- fotosyntéza * účinky záření MeSH
- fotosystém II - proteinový komplex metabolismus MeSH
- fykokyanin * metabolismus MeSH
- karotenoidy metabolismus MeSH
- peroxidace lipidů účinky záření MeSH
- reaktivní formy kyslíku * metabolismus MeSH
- Spirulina * účinky záření metabolismus MeSH
- ultrafialové záření * MeSH
- Publikační typ
- časopisecké články MeSH
Dunaliella salina is a rich source of carotenoids. Carotenoid production is induced under specific conditions, i.e., high light intensity, high salt concentration, nutrient limitation, and suboptimal temperatures in this microalga. The control of environmental factors is vital for high productivity of carotenoids. In this paper, the effect of different ethanol concentrations in combination with nitrogen deficiency was investigated to induce carotenoid production in D. salina CCAP 19/18. Also, some biochemical and molecular parameters were investigated in response to ethanol in the cells. It was shown that ethanol at 0.5% concentration increased cell number but, at 5% concentration, reduced cell viability compared to the control. The highest carotenoid production was achieved at 3% ethanol concentration, which was 1.46 fold higher than the nitrogen deficiency condition. Investigation of the 3 carotenoid biosynthesis genes revealed that their expression levels increased at 3% ethanol concentration, and the phytoene synthase gene was the most upregulated one. Lipid peroxidation increased at both 3% and 5% ethanol concentrations. At 3% concentration, the activity of catalase and superoxide dismutase increased, but no significant changes were seen at 5% ethanol concentration. Peroxidase activity reduced at both 3% and 5% concentrations. Moreover, proline and reducing sugar content increased at 3% concentration while decreased at 5% ethanol concertation. The results showed that at 3% ethanol concentration, higher carotenoid productivity was associated with an increase in other intracellular responses (molecular and biochemical). Ethanol as a controllable element may be beneficial to increase carotenoid production even under inappropriate environmental conditions in D. salina.
- MeSH
- antioxidancia * MeSH
- dusík MeSH
- karotenoidy * metabolismus MeSH
- superoxiddismutasa metabolismus MeSH
- světlo MeSH
- Publikační typ
- časopisecké články MeSH
Improving yield, nutritional value and tolerance to abiotic stress are major targets of current breeding and biotechnological approaches that aim at increasing crop production and ensuring food security. Metabolic engineering of carotenoids, the precursor of vitamin-A and plant hormones that regulate plant growth and response to adverse growth conditions, has been mainly focusing on provitamin A biofortification or the production of high-value carotenoids. Here, we show that the introduction of a single gene of the carotenoid biosynthetic pathway in different tomato cultivars induced profound metabolic alterations in carotenoid, apocarotenoid and phytohormones pathways. Alterations in isoprenoid- (abscisic acid, gibberellins, cytokinins) and non-isoprenoid (auxin and jasmonic acid) derived hormones together with enhanced xanthophyll content influenced biomass partitioning and abiotic stress tolerance (high light, salt, and drought), and it caused an up to 77% fruit yield increase and enhanced fruit's provitamin A content. In addition, metabolic and hormonal changes led to accumulation of key primary metabolites (e.g. osmoprotectants and antiaging agents) contributing with enhanced abiotic stress tolerance and fruit shelf life. Our findings pave the way for developing a new generation of crops that combine high productivity and increased nutritional value with the capability to cope with climate change-related environmental challenges.
Life on Earth depends on photosynthesis, the conversion of light energy into chemical energy. Plants collect photons by light harvesting complexes (LHC)-abundant membrane proteins containing chlorophyll and xanthophyll molecules. LHC-like proteins are similar in their amino acid sequence to true LHC antennae, however, they rather serve a photoprotective function. Whether the LHC-like proteins bind pigments has remained unclear. Here, we characterize plant LHC-like proteins (LIL3 and ELIP2) produced in the cyanobacterium Synechocystis sp. PCC 6803 (hereafter Synechocystis). Both proteins were associated with chlorophyll a (Chl) and zeaxanthin and LIL3 was shown to be capable of quenching Chl fluorescence via direct energy transfer from the Chl Qy state to zeaxanthin S1 state. Interestingly, the ability of the ELIP2 protein to quench can be acquired by modifying its N-terminal sequence. By employing Synechocystis carotenoid mutants and site-directed mutagenesis we demonstrate that, although LIL3 does not need pigments for folding, pigments stabilize the LIL3 dimer.
- MeSH
- chlorofyl metabolismus MeSH
- karotenoidy metabolismus MeSH
- multimerizace proteinu MeSH
- mutace MeSH
- přenos energie MeSH
- proteiny chloroplastové chemie genetika metabolismus MeSH
- proteiny huseníčku chemie genetika metabolismus MeSH
- sbalování proteinů MeSH
- Synechocystis genetika metabolismus MeSH
- vazba proteinů MeSH
- xanthofyly metabolismus MeSH
- zeaxanthiny genetika metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Light plays an essential role in photosynthesis; however, its excess can cause damage to cellular components. Photosynthetic organisms thus developed a set of photoprotective mechanisms (e.g., non-photochemical quenching, photoinhibition) that can be studied by a classic biochemical and biophysical methods in cell suspension. Here, we combined these bulk methods with single-cell identification of microdomains in thylakoid membrane during high-light (HL) stress. We used Synechocystis sp. PCC 6803 cells with YFP tagged photosystem I. The single-cell data pointed to a three-phase response of cells to acute HL stress. We defined: (1) fast response phase (0-30 min), (2) intermediate phase (30-120 min), and (3) slow acclimation phase (120-360 min). During the first phase, cyanobacterial cells activated photoprotective mechanisms such as photoinhibition and non-photochemical quenching. Later on (during the second phase), we temporarily observed functional decoupling of phycobilisomes and sustained monomerization of photosystem II dimer. Simultaneously, cells also initiated accumulation of carotenoids, especially ɣ-carotene, the main precursor of all carotenoids. In the last phase, in addition to ɣ-carotene, we also observed accumulation of myxoxanthophyll and more even spatial distribution of photosystems and phycobilisomes between microdomains. We suggest that the overall carotenoid increase during HL stress could be involved either in the direct photoprotection (e.g., in ROS scavenging) and/or could play an additional role in maintaining optimal distribution of photosystems in thylakoid membrane to attain efficient photoprotection.
- MeSH
- bakteriální proteiny genetika metabolismus MeSH
- fotosystém I - proteinový komplex genetika metabolismus MeSH
- fotosystém II - proteinový komplex genetika metabolismus MeSH
- karotenoidy metabolismus MeSH
- světlo * MeSH
- Synechocystis metabolismus účinky záření MeSH
- tylakoidy metabolismus účinky záření MeSH
- velikost buňky účinky záření MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Here, we propose a possible photoactivation mechanism of a 35-kDa blue light-triggered photoreceptor, the Orange Carotenoid Protein (OCP), suggesting that the reaction involves the transient formation of a protonated ketocarotenoid (oxocarbenium cation) state. Taking advantage of engineering an OCP variant carrying the Y201W mutation, which shows superior spectroscopic and structural properties, it is shown that the presence of Trp201 augments the impact of one critical H-bond between the ketocarotenoid and the protein. This confers an unprecedented homogeneity of the dark-adapted OCP state and substantially increases the yield of the excited photoproduct S*, which is important for the productive photocycle to proceed. A 1.37 Å crystal structure of OCP Y201W combined with femtosecond time-resolved absorption spectroscopy, kinetic analysis, and deconvolution of the spectral intermediates, as well as extensive quantum chemical calculations incorporating the effect of the local electric field, highlighted the role of charge-transfer states during OCP photoconversion.
Technologies based on synthetic biology to produce bacterial natural carotenoids depend on information regarding their biosynthesis. Although the biosynthetic pathway of common carotenoids is known, there are carotenoids whose pathways are not completely described. This work aimed to mine the genome of the deep-sea bacterium Erythrobacter citreus LAMA 915, an uncommon bacterium that forms yellow colonies under cultivation. This work further explores the potential application of the carotenoids found and low-cost substrates for bacterial growth. A combined approach of genome mining and untargeted metabolomics analysis was applied. The carotenoid erythroxanthin sulfate was detected in E. citreus LAMA 915 cell extract. A proposal for carotenoid biosynthesis by this bacterium is provided, involving the genes crtBIYZWG. These are responsible for the biosynthesis of carotenoids from the zeaxanthin pathway and their 2,2'-hydroxylated derivatives. E. citreus LAMA 915 extracts showed antioxidant and sun protection effects. Based on the high content of proteases and lipases, it was possible to rationally select substrates for bacterial growth, with residual oil from fish processing the best low-cost substrate selected. This work advances in the understanding of carotenoid biosynthesis and provides a genetic basis that can be further explored as a biotechnological route for carotenoid production.
- MeSH
- biosyntetické dráhy * genetika MeSH
- karotenoidy * metabolismus MeSH
- Sphingomonadaceae * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
The aim of this study was to characterize carotenoids and flavonoids present in French marigold flowers (Tagetes patula "Durango Red") as well as to assess the effects of harvest dates and postharvest treatments on these compounds. Carotenoids and flavonoids were quantified using their respective standards or semiquantified using relevant related standards. Lutein and its derivatives largely dominated the flower carotenoids, while the flavonoids were diverse with patuletin leading the list. Of the different postharvest treatments, ensilage leads to the highest content of carotenoids (from 5.0 to 7.8 g kg-1 dw) and flavonoids (from 19 to 50 g kg-1 dw). The composition of individual flavonoids was greatly influenced by different postharvest treatments, while the influence of harvest dates was secondary. Ensilage and drying induced separate metabolic pathways leading to degradation of high-molecular glycosidic compounds, converting the compounds either to their aglycones or into the intermediate flavonoid glycosides. We conclude that according to the intended industrial application, different postharvest techniques can be employed to acquire desired flavonoids on a large scale.
- MeSH
- časové faktory MeSH
- flavonoidy chemie metabolismus MeSH
- karotenoidy chemie metabolismus MeSH
- květy chemie růst a vývoj metabolismus MeSH
- lutein chemie metabolismus MeSH
- rostlinné extrakty chemie metabolismus MeSH
- Tagetes chemie růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Lichens survive harsh weather of Antarctica as well as of other hostile environments worldwide. Therefore, this investigation is important to understand the evolution of life on Earth in relation to their stress tolerance strategy. We have used chlorophyll a fluorescence (ChlF) and Raman spectroscopy, respectively, to monitor the activation/deactivation of photosynthesis and carotenoids in three diverse Antarctic lichens, Dermatocarpon polyphyllizum (DP), Umbilicaria antarctica (UA), and Leptogium puberulum (LP). These lichens, post 4 h or 24 h of hydration, showed differences in their ChlF transients and values of major ChlF parameters, e.g., in the maximum quantum efficiency of PSII photochemistry (Fv/Fm), and yields of fluorescence and heat dissipation (Φf,d), of effective quantum efficiency of PSII photochemistry (ΦPSII) and of non-photochemical quenching (Φnpq), which may be due to quantitative and/or qualitative differences in the composition of their photobionts. For understanding the kinetics of hydration-induced activation of photosynthesis, we screened ΦPSII of these lichens and reported its non-linear stimulation on a minute time scale; half of the activation time (t1/2) was fastest ~4.05 ± 0.29 min for DP, which was followed by 5.46 ± 0.18 min for UA, and 13.95 ± 1.24 min for LP. Upon drying of fully activated lichen thallus, there was a slow decay, in hours, of relative water content (RWC) as well as of Fv/Fm. Raman spectral signatures were different for lichens having algal (in DP and UA) and cyanobacteria (in LP) photobionts, and there was a significant shift in ν1(C=C) Raman band of carotenoids post 24 h hydration as compared to their value at a dry state or post 4 h of hydration; this shift was decreased, when drying, in DP and LP but not in UA. We conclude that hydration nonlinearly activated photosynthetic apparatus/reactions of these lichens in minute time range but there was a de-novo synthesis of chlorophylls as well as of carotenoids post 24 h. Their dehydration-induced deactivation, however, was comparatively slow, in hours range, and there seemed a degradation of synthesized chlorophylls and carotenoids post dryness. We conclude that in extremophilic lichens, their photosynthetic partners, in particular, possess a complex survival and photoprotective strategy to be successful in the extreme terrestrial environments in Antarctica.
- MeSH
- Ascomycota MeSH
- chlorofyl a MeSH
- chlorofyl MeSH
- fluorescence MeSH
- fotosyntéza MeSH
- karotenoidy metabolismus MeSH
- lišejníky * metabolismus MeSH
- Ramanova spektroskopie MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Antarktida MeSH
The effects of combining naturally evolved photosynthetic pigment-protein complexes with inorganic functional materials, especially plasmonically active metallic nanostructures, have been a widely studied topic in the last few decades. Besides other applications, it seems to be reasonable using such hybrid systems for designing future biomimetic solar cells. In this paper, we describe selected results that point out to various aspects of the interactions between photosynthetic complexes and plasmonic excitations in Silver Island Films (SIFs). In addition to simple light-harvesting complexes, like peridinin-chlorophyll-protein (PCP) or the Fenna-Matthews-Olson (FMO) complex, we also discuss the properties of large, photosynthetic reaction centers (RCs) and Photosystem I (PSI)-both prokaryotic PSI core complexes and eukaryotic PSI supercomplexes with attached antenna clusters (PSI-LHCI)-deposited on SIF substrates.
- MeSH
- chlorofyl a metabolismus MeSH
- fluorescenční spektrometrie metody MeSH
- formaldehyd chemie MeSH
- fotosyntéza * MeSH
- fotosystém I - proteinový komplex metabolismus MeSH
- glukosa chemie MeSH
- karotenoidy metabolismus MeSH
- nanostruktury chemie ultrastruktura MeSH
- stříbro chemie MeSH
- světlosběrné proteinové komplexy metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
