Irradiance and nutrient-dependent effects on photosynthetic electron transport in Arctic phytoplankton: A comparison of two chlorophyll fluorescence-based approaches to derive primary photochemistry

We employed Fast Repetition Rate fluorometry for high-resolution mapping of marine phytoplankton photophysiology and primary photochemistry in the Lancaster Sound and Barrow Strait regions of the Canadian Arctic Archipelago in the summer of 2019. Continuous ship-board analysis of chlorophyll a variable fluorescence demonstrated relatively low photochemical efficiency over most of the cruise-track, with the exception of localized regions within Barrow Strait, where there was increased vertical mixing and proximity to land-based nutrient sources. Along the full transect, we observed strong non-photochemical quenching of chlorophyll fluorescence, with relaxation times longer than the 5-minute period used for dark acclimation. Such long-term quenching effects complicate continuous underway acquisition of fluorescence amplitude-based estimates of photosynthetic electron transport rates, which rely on dark acclimation of samples. As an alternative, we employed a new algorithm to derive electron transport rates based on analysis of fluorescence relaxation kinetics, which does not require dark acclimation. Direct comparison of kinetics- and amplitude-based electron transport rate measurements demonstrated that kinetic-based estimates were, on average, 2-fold higher than amplitude-based values. The magnitude of decoupling between the two electron transport rate estimates increased in association with photophysiological diagnostics of nutrient stress. Discrepancies between electron transport rate estimates likely resulted from the use of different photophysiological parameters to derive the kinetics- and amplitude-based algorithms, and choice of numerical model used to fit variable fluorescence curves and analyze fluorescence kinetics under actinic light. Our results highlight environmental and methodological influences on fluorescence-based photochemistry estimates, and prompt discussion of best-practices for future underway fluorescence-based efforts to monitor phytoplankton photosynthesis.

Using Chlorophyll Fluorescence to Determine the Fate of Photons Absorbed by Phytoplankton in the World's Oceans

Approximately 45% of the photosynthetically fixed carbon on Earth occurs in the oceans in phytoplankton, which account for less than 1% of the world's photosynthetic biomass. This amazing empirical observation implies a very high photosynthetic energy conversion efficiency, but how efficiently is the solar energy actually used? The photon energy budget of photosynthesis can be divided into three terms: the quantum yields of photochemistry, fluorescence, and heat. Measuring two of these three processes closes the energy budget. The development of ultrasensitive, seagoing chlorophyll variable fluorescence and picosecond fluorescence lifetime instruments has allowed independent closure on the first two terms. With this closure, we can understand how phytoplankton respond to nutrient supplies on timescales of hours to months and, over longer timescales, to changes in climate. Expected final online publication date for the Annual Review of Marine Science, Volume 14 is January 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

Chlorophyll fluorescence-based estimates of photosynthetic electron transport in Arctic phytoplankton assemblages

We employed Fast Repetition Rate fluorometry for high-resolution mapping of marine phytoplankton photophysiology and primary productivity in the Lancaster Sound and Barrow Strait regions of the Canadian Arctic Archipelago in the summer of 2019. Continuous ship-board analysis of chlorophyll a variable fluorescence demonstrated relatively low photochemical efficiency over most of the cruise-track, with the exception of localized regions within Barrow Strait where there was increased vertical mixing and proximity to land-based nutrient sources. Along the full transect, we observed strong non-photochemical quenching of chlorophyll fluorescence, with relaxation times longer than the 5-minute period used for dark acclimation. Such long-term quenching effects complicate continuous underway acquisition of fluorescence amplitude-based estimates of photosynthetic electron transport rates, which rely on dark acclimation of samples. As an alternative, we employed a new algorithm to derive electron transport rates based on analysis of fluorescence relaxation kinetics, which does not require dark acclimation. Direct comparison of kinetics- and amplitude-based electron transport rate measurements demonstrated kinetic-based estimates were, on average, 2-fold higher than amplitude-based values. The magnitude of decoupling between the two electron transport rate estimates increased in association with photophysiological diagnostics of nutrient stress. Discrepancies between electron transport rate estimates likely resulted from the use of different photophysiological parameters to derive the kinetics- and amplitude-based algorithms, and choice of numerical model used to fit variable fluorescence curves and analyze fluorescence kinetics under actinic light. Our results highlight environmental and methodological influences on fluorescence-based productivity estimates, and prompt discussion of best-practices for future underway fluorescence-based efforts to monitor phytoplankton photosynthesis.

Ascorbic Acid-Induced Photosynthetic Adaptability of Processing Tomatoes to Salt Stress Probed by Fast OJIP Fluorescence Rise

In this study, the protective role of exogenous ascorbic acid (AsA) on salt-induced inhibition of photosynthesis in the seedlings of processing tomatoes under salt stress has been investigated. Plants under salt stress (NaCl, 100 mmol/L) were foliar-sprayed with AsA (0.5 mmol/L), lycorine (LYC, 0.25 mmol/L, an inhibitor of key AsA synthesis enzyme l-galactono-γ-lactone dehydrogenase activity), or AsA plus LYC. The effects of AsA on fast OJIP fluorescence rise curve and JIP parameters were then examined. Our results demonstrated that applying exogenous AsA significantly changed the composition of O-J-I-P fluorescence transients in plants subjected to salt stress both with and without LYC. An increase in basal fluorescence (Fo) and a decrease in maximum fluorescence (Fm) were observed. Lower K- and L-bands and higher I-band were detected on the OJIP transient curves compared, respectively, with salt-stressed plants with and without LYC. AsA application also significantly increased the values of normalized total complementary area (Sm), relative variable fluorescence intensity at the I-step (VI), absorbed light energy (ABS/CSm), excitation energy (TRo/CSm), and reduction energy entering the electron transfer chain beyond QA (ETo/CSm) per reaction centre (RC) and electron transport flux per active RC (ETo/RC), while decreasing some others like the approximated initial slope of the fluorescence transient (Mo), relative variable fluorescence intensity at the K-step (VK), average absorption (ABS/RC), trapping (TRo/RC), heat dissipation (DIo/RC) per active RC, and heat dissipation per active RC (DIo/CSm) in the presence or absence of LYC. These results suggested that exogenous AsA counteracted salt-induced photoinhibition mainly by modulating the endogenous AsA level and redox state in the chloroplast to promote chlorophyll synthesis and alleviate the damage of oxidative stress to photosynthetic apparatus. AsA can also raise the efficiency of light utilization as well as excitation energy dissipation within the photosystem II (PSII) antennae, thus increasing the stability of PSII and promoting the movement of electrons among PS1 and PSII in tomato seedling leaves subjected to salt stress.

Inhibition of Primary Photosynthesis in Desiccating Antarctic Lichens Differing in Their Photobionts, Thallus Morphology, and Spectral Properties

Five macrolichens of different thallus morphology from Antarctica (King George Island) were used for this ecophysiological study. The effect of thallus desiccation on primary photosynthetic processes was examined. We investigated the lichens’ responses to the relative water content (RWC) in their thalli during the transition from a wet (RWC of 100%) to a dry state (RWC of 0%). The slow Kautsky kinetics of chlorophyll fluorescence (ChlF) that was recorded during controlled dehydration (RWC decreased from 100 to 0%) and supplemented with a quenching analysis revealed a polyphasic species-specific response of variable fluorescence. The changes in ChlF at a steady state (Fs), potential and effective quantum yields of photosystem II (FV/FM, ΦPSII), and nonphotochemical quenching (NPQ) reflected a desiccation-induced inhibition of the photosynthetic processes. The dehydration-dependent fall in FV/FM and ΦPSII was species-specific, starting at an RWC range of 22–32%. The critical RWC for ΦPSII was below 5%. The changes indicated the involvement of protective mechanisms in the chloroplastic apparatus of lichen photobionts at RWCs of below 20%. In both the wet and dry states, the spectral reflectance curves (SRC) (wavelength 400–800 nm) and indices (NDVI, PRI) of the studied lichen species were measured. Black Himantormia lugubris showed no difference in the SRCs between wet and dry state. Other lichens showed a higher reflectance in the dry state compared to the wet state. The lichen morphology and anatomy data, together with the ChlF and spectral reflectance data, are discussed in relation to its potential for ecophysiological studies in Antarctic lichens.

Growth and Physiological Responses of Norway Spruce (Picea abies (L.) H. Karst) Supplemented with Monochromatic Red, Blue and Far-Red Light

Monochromatic red light (R) supplementation is more efficient than blue light (B) in promoting Norway spruce (Picea abies (L.) H. Karst) growth. Transcriptome analysis has revealed that R and B may regulate stem growth by regulating phytohormones and secondary metabolites; however, the effects of light qualities on physiological responses and related gene expression in Norway spruce require further study. In the present study, three-year-old Norway spruce seedlings received sunlight during the daytime were exposed to monochromatic B (460 mm), monochromatic R (660 nm), monochromatic far-red light (FR, 730 nm), and a combination of three monochromatic lights (control, R:FR:B = 7:1:1) using light-emitting diode (LED) lamps for 12 h after sunset for 90 day. Growth traits, physiological responses, and related gene expression were determined. The results showed that light quality significantly affected Norway spruce growth. The stem height, root collar diameter, and current-year shoot length of seedlings treated with R were 2%, 10% and 12% higher, respectively, than those of the control, whereas seedlings treated with B and FR showed significantly lower values of these parameters compared with that of the control. The net photosynthetic rate (Pn) of seedlings under R treatment was 10% higher than that of the control, whereas the Pn values of seedlings treated with FR and B were 22% and 33%, respectively, lower than that of the control. The ratio of phosphoenolpyruvate carboxylase to ribulose-1,5-bisphosphate carboxylase/oxygenase (PEPC/Rubisco) of seedlings after the R treatment (0.581) was the highest and 3.98 times higher than that of the seedlings treated with B. Light quality significantly affected the gibberellic acid (GAs) levels, which was 13% higher in seedlings treated with R (6.4 g/100 ng) than that of the control, whereas, the GAs level of seedlings treated with B and FR was 17% and 19% lower, respectively, than that of the control. In addition, seedlings treated with R achieved the lowest ratio of leaf chlorophyll content to fresh weight (8.7). Compared to the R and control treatments, seedlings received FR treatment had consistently lower values of the quantum yield of electron transport beyond QA− (primary quinone, ϕEo) and efficiency, with which a trapped exciton moves an electron into the electron transport chain beyond QA− (ψo), while higher values of the relatively variable fluorescence at the J step and normalized relatively variable fluorescence at the K step (Wk). The values of ϕEo, ψO, VJ and Wk in seedlings treated with B were similar to those in the control group. The expression of genes associated with light signal transduction, such as PHYTOCHROME C (PHYC), ELONGATED HYPOCOTYL5 (HY5), CONSTITUTIVE PHOTOMORPHOGENIC 1-2 (COP1-2), and PHYTOCHROMEINTERACTING FACTOR 3 (PIF3), was significantly higher in seedlings under B treatment than those under other light treatments. Nevertheless, significant differences were not observed in the expression of COP1-2, HY5, and PIF3 between the R treatment and the control. The expression value of COP1-2 was significantly lower in R than FR light treatments. In conclusion, compared with the control, R promotes, whereas B and FR inhibit Norway spruce growth, which was accompanied by physiological changes and genes expression regulation that may be relate to a changing phytochrome photostationary state (PSS) with the supplemental R in seedlings.

Evidence for variable chlorophyll fluorescence of photosystem I in vivo

Room temperature fluorescence in vivo and its light-induced changes are dominated by chlorophyll a fluorescence excited in photosystem II, F(II), peaking around 685 nm. Photosystem I fluorescence, F(I), peaking around 730 nm, so far has been assumed to be constant in vivo. Here, we present evidence for significant contributions of F(I) to variable fluorescence in the green unicellular alga Chlorella vulgaris, the cyanobacterium Synechococcus leopoliensis and a light-green ivy leaf. A Multi-Color-PAM fluorometer was applied for measurements of the polyphasic fluorescence rise (O-I1-I2-P) induced by strong 440 nm light in a dilute suspension of Chlorella, with detection alternating between emission above 700 nm (F > 700) and below 710 nm (F < 710). By averaging 10 curves each of the F > 700 and F < 710 recordings even small differences could be reliably evaluated. After equalizing the amplitudes of the O-I1 phase, which constitutes a specific F(II) response, the O-I1-I2 parts of the two recordings were close to identical, whereas the I2-P phase was larger in F > 700 than in F < 710 by a factor of 1.42. In analogous measurements with Synechococcus carried out in the dark state 2 using strong 625 nm actinic light, after O-I1 equalization the I2-P phase in F > 700 exceeded that in F < 710 even by a factor of 1.99. In measurements with Chlorella, the I2-P phase and with it the apparent variable fluorescence of PS I, Fv(I), were suppressed by moderate actinic background light and by the plastoquinone antagonist DBMIB. Analogous measurements with leaves are rendered problematic by unavoidable light intensity gradients and the resulting heterogenic origins of F > 700 and F < 710. However, a light-green young ivy leaf gave qualitatively similar results as those obtained with the suspensions, thus strongly suggesting the existence of Fv(I) also in leaves.

Chloroplast pigments and photochemical efficiency of West Indian cherry under salt stress and potassium-phosphorus fertilization

The objective of this study was to evaluate chloroplast pigments and photochemical efficiency of West Indian cherry cv. BRS 366 Jaburu as a function of irrigation water salinity and potassium-phosphorus fertilization combinations in the second year of cultivation. The experiment was carried out in a protected environment in Campina Grande, Brazil. Treatments were distributed in randomized blocks, in a 5 × 4 factorial scheme, corresponding to five levels of electrical conductivity of irrigation water ECw (0.6, 1.4, 2.2, 3.0 and 3.8 dS m-1) and four combinations of potassium-phosphorus fertilization (100/100, 85/85, 60/60 and 45/45% of the K2O/P2O5 recommendation for the second year of cultivation 200 g of K2O and 120 g of P2O5 per plant per year) with three replicates. Irrigation with saline waters hampered the biosynthesis of chloroplast pigments and the photochemical efficiency of West Indian cherry cv. BRS 366 Jaburu in the second year of cultivation. Water salinity from 2.6 dS m-1 reduced the maximum fluorescence, variable fluorescence, and quantum efficiency of photosystem II of West Indian cherry plants cv. BRS 366 Jaburu. Fertilization with 60/60 and 85/85% of the K2O/P2O5 recommendation promotes an increase in the synthesis of chlorophylls a and b, respectively, in the first and second production cycles of the second year of cultivation. Supply of 85/85% of the K2O/P2O5 recommendation promoted an increase in maximum and variable fluorescence in plants subjected to water salinities of 0.6, 2.2, and 3.8 dS m-1 in the second cycle and reduced the initial fluorescence, regardless of the salinity level in the first and second production cycles of West Indian cherry.