Three basil plant varieties (Ocimum basilicum var. Genovese, Ocimum × citriodorum, and Ocimum basilicum var. purpurascens) were grown under moderate light (about 300 μmol photons m-2 s-1) in a glasshouse or growth chamber and then either transferred to an open field (average daily dose: 29.2 kJ m-2 d-1) or additionally exposed to UV-B irradiation in a growth chamber (29.16 kJ m-2 d-1), to reveal the variety-specific and light-specific acclimation responses. Total antioxidant capacity (TAC), phenolic profile, ascorbate content, and class III peroxidase (POD) activity were used to determine the antioxidant status of leaves under all four light regimes. Exposure to high solar irradiation at the open field resulted in an increase in TAC, total hydroxycinnamic acids (HCAs, especially caffeic acid), flavonoids, and epidermal UV-absorbing substances in all three varieties, as well as a two-fold increase in the leaf dry/fresh weight ratio. The supplemental UV-B irradiation induced preferential accumulation of HCAs (rosmarinic acid) over flavonoids, increased TAC and POD activity, but decreased the ascorbate content in the leaves, and inhibited the accumulation of epidermal flavonoids in all basil varieties. Furthermore, characteristic leaf curling and UV-B-induced inhibition of plant growth were observed in all basil varieties, while a pro-oxidant effect of UV-B was indicated with H2O2 accumulation in the leaves and spotty leaf browning. The extent of these morphological changes, and oxidative damage depended on the basil cultivar, implies a genotype-specific tolerance mechanism to high doses of UV-B irradiation.
- MeSH
- Antioxidants * pharmacology MeSH
- Ocimum basilicum * MeSH
- Flavonoids MeSH
- Ascorbic Acid MeSH
- Plant Leaves MeSH
- Hydrogen Peroxide MeSH
- Sunlight MeSH
- Publication type
- Journal Article 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
- Bacterial Proteins genetics metabolism MeSH
- Photosystem I Protein Complex genetics metabolism MeSH
- Photosystem II Protein Complex genetics metabolism MeSH
- Carotenoids metabolism MeSH
- Light * MeSH
- Synechocystis metabolism radiation effects MeSH
- Thylakoids metabolism radiation effects MeSH
- Cell Size radiation effects MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Light quality significantly influences plant metabolism, growth and development. Recently, we have demonstrated that leaves of barley and other plant species grown under monochromatic green light (500-590 nm) accumulated a large pool of chlorophyll a (Chl a) intermediates with incomplete hydrogenation of their phytyl chains. In this work, we studied accumulation of these geranylgeranylated Chls a and b in pigment-protein complexes (PPCs) of Arabidopsis plants acclimated to green light and their structural-functional consequences on the photosynthetic apparatus. We found that geranylgeranylated Chls are present in all major PPCs, although their presence was more pronounced in light-harvesting complex II (LHCII) and less prominent in supercomplexes of photosystem II (PSII). Accumulation of geranylgeranylated Chls hampered the formation of PSII and PSI super- and megacomplexes in the thylakoid membranes as well as their assembly into chiral macrodomains; it also lowered the temperature stability of the PPCs, especially that of LHCII trimers, which led to their monomerization and an anomaly in the photoprotective mechanism of non-photochemical quenching. Role of geranylgeranylated Chls in adverse effects on photosynthetic apparatus of plants acclimated to green light is discussed.
Plant survival in temperate zones requires efficient cold acclimation, which is strongly affected by light and temperature signal crosstalk, which converge in modulation of hormonal responses. Cold under low light conditions affected Arabidopsis responses predominantly in apices, possibly because energy supplies were too limited for requirements of these meristematic tissues, despite a relatively high steady-state quantum yield. Comparing cold responses at optimal light intensity and low light, we found activation of similar defence mechanisms-apart from CBF1-3 and CRF3-4 pathways, also transient stimulation of cytokinin type-A response regulators, accompanied by fast transient increase of trans-zeatin in roots. Upregulated expression of components of strigolactone (and karrikin) signalling pathway indicated involvement of these phytohormones in cold responses. Impaired response of phyA, phyB, cry1 and cry2 mutants reflected participation of these photoreceptors in acquiring freezing tolerance (especially cryptochrome CRY1 at optimal light intensity and phytochrome PHYA at low light). Efficient cold acclimation at optimal light was associated with upregulation of trans-zeatin in leaves and roots, while at low light, cytokinin (except cis-zeatin) content remained diminished. Cold stresses induced elevation of jasmonic acid and salicylic acid (in roots). Low light at optimal conditions resulted in strong suppression of cytokinins, jasmonic and salicylic acid.
The benefits of in vitro plant cultivation are mainly due to very high multiplication rate. Cultivation of plant material in vitro can be carried out during the whole year regardless of the time of the year or weather conditions. We create artificial conditions in the lab (heat, light, humidity), and we can regulate these conditions at any time. For the preservation of cultivar identity, we recommend establishing in vitro cultures from shoot tips usually larger than 0.2 mm. In practice, in vitro cultivation of plants uses these growth regulators to achieve organogenesis, for example, root formation, prolonged growth, or multiplication. During each subculture, these cultures are then transferred on a solid agar medium in the form of actively growing multiple shoots with a well-differentiated shoot tip containing meristematic area. Cytokinins are important for cell division and causes branching of plants. Auxins, both endogenous and exogenous, act at as a trigger for the differentiation and formation of root primordia. Morphological characteristics (formation of leaves or callus) and shoot development should be observed during in vitro multiplication and after transfer to ex vitro conditions.
- MeSH
- Acclimatization physiology MeSH
- Cytokinins pharmacology MeSH
- Culture Media chemistry pharmacology MeSH
- Indoleacetic Acids pharmacology MeSH
- Meristem drug effects growth & development MeSH
- Cell Proliferation drug effects physiology MeSH
- Plant Growth Regulators pharmacology MeSH
- Rosaceae drug effects growth & development MeSH
- In Vitro Techniques MeSH
- Tissue Culture Techniques methods MeSH
- Plant Shoots drug effects growth & development MeSH
- Publication type
- Journal Article MeSH
BACKGROUND AND AIMS: UVB radiation can rapidly induce gene regulation leading to cumulative changes for plant physiology and morphology. We hypothesized that a transgenerational effect of chronic exposure to solar short UV modulates the offspring's responses to UVB and blue light, and that the transgenerational effect is genotype dependent. METHODS: We established a factorial experiment combining two Vicia faba L. accessions, two parental UV treatments (full sunlight and exclusion of short UV, 290-350 nm), and four offspring light treatments from the factorial combination of UVB and blue light. The accessions were Aurora from southern Sweden, and ILB938 from Andean region of Colombia and Ecuador. KEY RESULTS: The transgenerational effect influenced morphological responses to blue light differently in the two accessions. In Aurora, when UVB was absent, blue light increased shoot dry mass only in plants whose parents were protected from short UV. In ILB938, blue light increased leaf area and shoot dry mass more in plants whose parents were exposed to short UV than those that were not. Moreover, when the offspring was exposed to UVB, the transgenerational effect decreased in ILB938 and disappeared in Aurora. For flavonoids, the transgenerational effect was detected only in Aurora: parental exposure to short UV was associated with a greater induction of total quercetin in response to UVB. Transcript abundance was higher in Aurora than in ILB938 for both CHALCONE SYNTHASE (99-fold) and DON-GLUCOSYLTRANSFERASE 1 (19-fold). CONCLUSIONS: The results supported both hypotheses. Solar short UV had transgenerational effects on progeny responses to blue and UVB radiation, and they differed between the accessions. These transgenerational effects could be adaptive by acclimation of slow and cumulative morphological change, and by early build-up of UV protection through flavonoid accumulation on UVB exposure. The differences between the two accessions aligned with their adaptation to contrasting UV environments.
- MeSH
- Adaptation, Physiological MeSH
- Sunlight * MeSH
- Ultraviolet Rays * MeSH
- Vicia faba genetics growth & development physiology MeSH
- Publication type
- Journal Article MeSH
Chlorophyll fluorescence kinetic analysis has become an important tool in basic and applied research on plant physiology and agronomy. While early systems recorded the integrated kinetics of a selected spot or plant, later systems enabled imaging of at least the slower parts of the kinetics (20-ms time resolution). For faster events, such as the rise from the basic dark-adapted fluorescence yield to the maximum (OJIP transient), or the fluorescence yield decrease during reoxidation of plastoquinone A after a saturating flash, integrative systems are used because of limiting speed of the available imaging systems. In our new macroscopic and microscopic systems, the OJIP or plastonique A reoxidation fluorescence transients are directly imaged using an ultrafast camera. The advantage of such systems compared to nonimaging measurements is the analysis of heterogeneity of measured parameters, for example between the photosynthetic tissue near the veins and the tissue further away from the veins. Further, in contrast to the pump-and-probe measurement, direct imaging allows for measuring the transition of the plant from the dark-acclimated to a light-acclimated state via a quenching analysis protocol in which every supersaturating flash is coupled to a measurement of the fast fluorescence rise. We show that pump-and-probe measurement of OJIP is prone to artifacts, which are eliminated with the direct measurement. The examples of applications shown here, zinc deficiency and cadmium toxicity, demonstrate that this novel imaging platform can be used for detection and analysis of a range of alterations of the electron flow around PSII.
- MeSH
- Arabidopsis cytology metabolism MeSH
- Brassicaceae cytology drug effects metabolism MeSH
- Chlorophyll chemistry metabolism MeSH
- Equipment Design MeSH
- Fluorescence MeSH
- Microscopy, Fluorescence instrumentation methods MeSH
- Photosynthesis MeSH
- Glycine max cytology drug effects metabolism MeSH
- Kinetics MeSH
- Plant Leaves cytology MeSH
- Mesophyll Cells metabolism MeSH
- Plastoquinone metabolism MeSH
- Zinc metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
The membrane-embedded FtsH proteases found in bacteria, chloroplasts, and mitochondria are involved in diverse cellular processes including protein quality control and regulation. The genome of the model cyanobacterium Synechocystis sp PCC 6803 encodes four FtsH homologs designated FtsH1 to FtsH4. The FtsH3 homolog is present in two hetero-oligomeric complexes: FtsH2/3, which is responsible for photosystem II quality control, and the essential FtsH1/3 complex, which helps maintain Fe homeostasis by regulating the level of the transcription factor Fur. To gain a more comprehensive insight into the physiological roles of FtsH hetero-complexes, we performed genome-wide expression profiling and global proteomic analyses of Synechocystis mutants conditionally depleted of FtsH3 or FtsH1 grown under various nutrient conditions. We show that the lack of FtsH1/3 leads to a drastic reduction in the transcriptional response to nutrient stress of not only Fur but also the Pho, NdhR, and NtcA regulons. In addition, this effect is accompanied by the accumulation of the respective transcription factors. Thus, the FtsH1/3 complex is of critical importance for acclimation to iron, phosphate, carbon, and nitrogen starvation in Synechocystis.plantcell;31/12/2912/FX1F1fx1.
- MeSH
- Acclimatization genetics MeSH
- Bacterial Proteins genetics metabolism MeSH
- Nitrogen deficiency metabolism MeSH
- Gene Expression MeSH
- Phosphates deficiency metabolism MeSH
- Phosphorylation MeSH
- Photosystem II Protein Complex chemistry genetics metabolism MeSH
- Metalloproteases genetics metabolism MeSH
- Mutation MeSH
- Phosphate-Binding Proteins genetics metabolism MeSH
- Proteolysis MeSH
- Proteome genetics metabolism MeSH
- Proteomics MeSH
- Gene Expression Regulation, Bacterial genetics MeSH
- Regulon genetics MeSH
- Repressor Proteins genetics metabolism MeSH
- Ribosomal Proteins genetics metabolism MeSH
- Synechocystis enzymology metabolism MeSH
- Transcription Factors genetics metabolism MeSH
- Carbon deficiency metabolism MeSH
- Nutrients deficiency metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Cold acclimation modifies the photosynthetic machinery and enables plants to survive at sub-zero temperatures, whereas in warm habitats, many species suffer even at non-freezing temperatures. We have measured chlorophyll a fluorescence (ChlF) and CO2 assimilation to investigate the effects of cold acclimation, and of low temperatures, on a cold-sensitive Arabidopsis thaliana accession C24. Upon excitation with low intensity (40 µmol photons m- 2 s- 1) ~ 620 nm light, slow (minute range) ChlF transients, at ~ 22 °C, showed two waves in the SMT phase (S, semi steady-state; M, maximum; T, terminal steady-state), whereas CO2 assimilation showed a linear increase with time. Low-temperature treatment (down to - 1.5 °C) strongly modulated the SMT phase and stimulated a peak in the CO2 assimilation induction curve. We show that the SMT phase, at ~ 22 °C, was abolished when measured under high actinic irradiance, or when 3-(3, 4-dichlorophenyl)-1, 1- dimethylurea (DCMU, an inhibitor of electron flow) or methyl viologen (MV, a Photosystem I (PSI) electron acceptor) was added to the system. Our data suggest that stimulation of the SMT wave, at low temperatures, has multiple reasons, which may include changes in both photochemical and biochemical reactions leading to modulations in non-photochemical quenching (NPQ) of the excited state of Chl, "state transitions," as well as changes in the rate of cyclic electron flow through PSI. Further, we suggest that cold acclimation, in accession C24, promotes "state transition" and protects photosystems by preventing high excitation pressure during low-temperature exposure.
A proper spatial distribution of photosynthetic pigment-protein complexes - PPCs (photosystems, light-harvesting antennas) is crucial for photosynthesis. In plants, photosystems I and II (PSI and PSII) are heterogeneously distributed between granal and stromal thylakoids. Here we have described similar heterogeneity in the PSI, PSII and phycobilisomes (PBSs) distribution in cyanobacteria thylakoids into microdomains by applying a new image processing method suitable for the Synechocystis sp. PCC6803 strain with yellow fluorescent protein-tagged PSI. The new image processing method is able to analyze the fluorescence ratios of PPCs on a single-cell level, pixel per pixel. Each cell pixel is plotted in CIE1931 color space by forming a pixel-color distribution of the cell. The most common position in CIE1931 is then defined as protein arrangement (PA) factor with xy coordinates. The PA-factor represents the most abundant fluorescence ratio of PSI/PSII/PBS, the 'mode color' of studied cell. We proved that a shift of the PA-factor from the center of the cell-pixel distribution (the 'median' cell color) is an indicator of the presence of special subcellular microdomain(s) with a unique PSI/PSII/PBS fluorescence ratio in comparison to other parts of the cell. Furthermore, during a 6-h high-light (HL) treatment, 'median' and 'mode' color (PA-factor) of the cell changed similarly on the population level, indicating that such microdomains with unique PSI/PSII/PBS fluorescence were not formed during HL (i.e. fluorescence changed equally in the whole cell). However, the PA-factor was very sensitive in characterizing the fluorescence ratios of PSI/PSII/PBS in cyanobacterial cells during HL by depicting a 4-phase acclimation to HL, and their physiological interpretation has been discussed.
