Chlorophyll-protein Complexes Research Articles

A Dose-Dependent Effect of UV-328 on Photosynthesis: Exploring Light Harvesting and UV-B Sensing Mechanisms

Benzotriazole ultraviolet (UV) stabilizers (BUVs) have emerged as significant environmental contaminants, frequently detected in various ecosystems. While the toxicity of BUVs to aquatic organisms is well-documented, studies on their impact on plant life are scarce. Plants are crucial as they provide the primary source of energy and organic matter in ecosystems through photosynthesis. This study investigated the effects of UV-328 (2-(2-hydroxy-4′,6′-di-tert-amylphenyl) benzotriazole) on plant growth indices and photosynthesis processes, employing conventional physiological experiments, RNA sequencing (RNA-seq) analysis, and computational methods. Results demonstrated a biphasic response in plant biomass and the maximum quantum yield of PS II (Fv/Fm), showing improvement at a 50μM UV-328 treatment but reduction under 150μM UV-328 exposure. Additionally, disruption in thylakoid morphology was observed at the higher concentration. RNA-seq and qRT-PCR analysis identified key differentially expressed genes (light-harvesting chlorophyll-protein complex Ⅰ subunit A4, light-harvesting chlorophyll b-binding protein 3, UVR8, and curvature thylakoid 1A) related to photosynthetic light harvesting, UV-B sensing, and chloroplast structure pathways, suggesting they may contribute to the observed alterations in photosynthesis activity induced by UV-328 exposure. Molecular docking analyses further supported the binding affinity between these proteins and UV-328. Overall, this study provided comprehensive physiological and molecular insights, contributing valuable information to the evaluation of the potential risks posed by UV-328 to critical plant physiological processes.

TaCAP1 interacts with TaLHCB1s and positively regulates wheat resistance against stripe rust.

Actin filaments and their associated actin-binding proteins (ABPs) play key roles in plant innate immune signaling. CAP1, or cyclase-associated protein 1, is an important regulatory factor of the actin cytoskeleton-associated signaling network, and was hypothesized here to be involved in resistance against wheat stripe rust because TaCAP1 expression was up-regulated in response to Puccinia striiformis f. sp. tritici (Pst). Down regulation of TaCAP1 expression led to decreased resistance against Pst, in contrast to increased resistance upon TaCAP1 overexpressing, as demonstrated by the changes of phenotypes and hyphal growth. We found increased expression of pathogenesis-responsive or relative related genes and disease grade changed in TaCAP1 overexpressing plants. Our results also showed TaCAP1-regulated host resistance to Pst by inducing the production and accumulation of reactive oxygen species (ROS) and mediating the salicylic acid signaling pathway. Additionally, TaCAP1 interacted with chlorophyll a/b -binding proteins TaLHCB1.3 and TaLHCB1.4, also known as light-harvesting chlorophyll-protein complex II subunit B (LHCB), which belong to the light-harvesting complex II proteins family. Silencing of two TaLHCB1 genes showed higher susceptibility to Pst, which reduced wheat resistance against Pst. Therefore, the data presented herein further illuminate our understanding that TaCAP1 interacts with TaLHCB1s, and functions as a positive regulator of wheat resistance against the stripe rust.

Mechanisms Underlying the Differential Sensitivity to Mesotrione in Sweet Corn

Mesotrione is a widely used post-emergence herbicide for maize. The toxicity of mesotrione to maize (especially sweet corn) has been widely reported, and some sweet corn varieties are highly sensitive to mesotrione, which affects subsequent plant growth periods. However, the molecular mechanisms responsible for the differences in susceptibility to mesotrione are not known. By comparing changes in the transcriptome of mesotrione-tolerant line 301 and mesotrione-sensitive line 276 after mesotrione treatment, we found that the genes coding light-harvesting chlorophyll protein complex were induced in 301, and the genes coding loosening cell walls were overrepresented in 276. The net photosynthetic rate, maximum photochemical efficiency of leaf PSII, photochemical quenching of chlorophyll fluorescence, and the electron transport rate were significantly higher in 301 than in 276 after mesotrione treatment, and these effects became more severe as time passed. In addition, oxidative balance was also affected by mesotrione. Compared with 301, SOD, POD, and CAT activities were significantly reduced with longer exposure time in 276. The results suggested that sweet corn can mitigate herbicide mesotrione toxicity by improving photosynthesis, ROS scavenging, and cell wall synthesis.

Dissection of the E8 locus in two early maturing Canadian soybean populations.

Soybean [Glycine max (L.) Merr.] is a short-day crop for which breeders want to expand the cultivation range to more northern agro-environments by introgressing alleles involved in early reproductive traits. To do so, we investigated quantitative trait loci (QTL) and expression quantitative trait loci (eQTL) regions comprised within the E8 locus, a large undeciphered region (~7.0 Mbp to 44.5 Mbp) associated with early maturity located on chromosome GM04. We used a combination of two mapping algorithms, (i) inclusive composite interval mapping (ICIM) and (ii) genome-wide composite interval mapping (GCIM), to identify major and minor regions in two soybean populations (QS15524F2:F3 and QS15544RIL) having fixed E1, E2, E3, and E4 alleles. Using this approach, we identified three main QTL regions with high logarithm of the odds (LODs), phenotypic variation explained (PVE), and additive effects for maturity and pod-filling within the E8 region: GM04:16,974,874-17,152,230 (E8-r1); GM04:35,168,111-37,664,017 (E8-r2); and GM04:41,808,599-42,376,237 (E8-r3). Using a five-step variant analysis pipeline, we identified Protein far-red elongated hypocotyl 3 (Glyma.04G124300; E8-r1), E1-like-a (Glyma.04G156400; E8-r2), Light-harvesting chlorophyll-protein complex I subunit A4 (Glyma.04G167900; E8-r3), and Cycling dof factor 3 (Glyma.04G168300; E8-r3) as the most promising candidate genes for these regions. A combinatorial eQTL mapping approach identified significant regulatory interactions for 13 expression traits (e-traits), including Glyma.04G050200 (Early flowering 3/E6 locus), with the E8-r3 region. Four other important QTL regions close to or encompassing major flowering genes were also detected on chromosomes GM07, GM08, and GM16. In GM07:5,256,305-5,404,971, a missense polymorphism was detected in the candidate gene Glyma.07G058200 (Protein suppressor of PHYA-105). These findings demonstrate that the locus known as E8 is regulated by at least three distinct genomic regions, all of which comprise major flowering genes.

Fluorescence quenching in aggregates of fucoxanthin–chlorophyll protein complexes: Interplay of fluorescing and dark states

Diatoms, a major group of algae, account for about a quarter of the global primary production on Earth. These photosynthetic organisms face significant challenges due to light intensity variations in their underwater habitat. To avoid photodamage, they have developed very efficient non-photochemical quenching (NPQ) mechanisms. These mechanisms originate in their light-harvesting antenna – the fucoxanthin–chlorophyll protein (FCP) complexes. Spectroscopic studies of NPQ in vivo are often hindered by strongly overlapping signals from the photosystems and their antennae. Fortunately, in vitro FCP aggregates constitute a useful model system to study fluorescence (FL) quenching in diatoms. In this work, we present streak-camera FL measurements on FCPa and FCPb complexes, isolated from a centric diatom Cyclotella meneghiniana, and their aggregates. We find that spectra of non-aggregated FCP are dominated by a single fluorescing species, but the FL spectra of FCP aggregates additionally contain contributions from a redshifted emissive state. We relate this red state to a charge transfer state between chlorophyll c and chlorophyll a molecules. The FL quenching, on the other hand, is due to an additional dark state that involves incoherent energy transfer to the fucoxanthin carotenoids. Overall, the global picture of energy transfer and quenching in FCP aggregates is very similar to that of major light-harvesting complexes in higher plants (LHCII), but microscopic details between FCPs and LHCIIs differ significantly.

Melatonin-Mediated Enhancement of Photosynthetic Capacity and Photoprotection Improves Salt Tolerance in Wheat

The role of melatonin in plant growth and response to environmental stress has been widely demonstrated. However, the physiological and molecular regulation of salt tolerance in wheat seedlings by melatonin remains unclear. In this study, we investigated changes in phenotype, physiology, photosynthetic parameters, and transcript levels in wheat seedlings to reveal the role of melatonin in the regulation of salt tolerance in wheat. The results indicate that the application of exogenous melatonin significantly alleviates growth inhibition, reactive oxygen species accumulation, and membrane oxidative damage induced by salt stress in wheat. Additionally, exogenous melatonin increased antioxidant enzyme activity and regulated photosynthetic gas exchange. Transcriptomic data showed a significant up-regulation of genes encoding light-harvesting chlorophyll protein complex proteins in photosynthesis and genes related to chlorophyll and carotenoid biosynthesis under the influence of melatonin. These results suggest that exogenous melatonin improves salt tolerance in wheat seedlings by enhancing the antioxidant, photoprotective, and photosynthesis activities.

Continuous Cropping Inhibits Photosynthesis of Polygonatum odoratum.

Polygonatum odoratum (Mill.) Druce possesses widespread medicinal properties; however, the continuous cropping (CC) often leads to a severe consecutive monoculture problem (CMP), ultimately causing a decline in yield and quality. Photosynthesis is the fundamental process for plant growth development. Improving photosynthesis is one of the most promising approaches to increase plant yields. To better understand how P. odoratum leaves undergo photosynthesis in response to CC, this study analyzed the physiochemical indexes and RNA-seq. The physiochemical indexes, such as the content of chlorophyll (chlorophyll a, b, and total chlorophyll), light response curves (LRCs), and photosynthetic parameters (Fv/Fm, Fv/F0, Fm/F0, Piabs, ABS/RC, TRo/RC, ETo/RC, and DIo/RC) were all changed in P. odoratum under the CC system. Furthermore, 13,798 genes that exhibited differential expression genes (DEGs) were identified in the P. odoratum leaves of CC and first cropping (FC) plants. Among them, 7932 unigenes were upregulated, while 5860 unigenes were downregulated. Here, the DEGs encoding proteins associated with photosynthesis and carbon assimilation showed a significant decrease in expression under the CC system, such as the PSII protein complex, PSI protein complex, Cytochorome b6/f complex, the photosynthetic electron transport chain, light-harvesting chlorophyll protein complex, and Calvin cycle, etc., -related gene. This study demonstrates that CC can suppress photosynthesis and carbon mechanism in P. odoratum, pinpointing potential ways to enhance photosynthetic efficiency in the CC of plants.

Combining BN-PAGE and microscopy techniques to investigate pigment-protein complexes and plastid transitions in citrus fruit

BackgroundChlorophyll and carotenoids, the most widely distributed lipophilic pigments in plants, contribute to fruit coloration during development and ripening. These pigments are assembled with pigment-protein complexes localized at plastid membrane. Pigment-protein complexes are essential for multiple cellular processes, however, their identity and composition in fruit have yet to be characterized.ResultsBy using BN-PAGE technique in combination with microscopy, we studied pigment-protein complexes and plastid transformation in the purified plastids from the exocarp of citrus fruit. The discontinuous sucrose gradient centrifugation was used to isolate total plastids from kumquat fruit, and the purity of isolated plastids was assessed by microscopy observation and western blot analysis. The isolated plastids at different coloring stages were subjected to pigment autofluorescence observation, western blot, two-dimensional electrophoresis analysis and BN-PAGE assessment. Our results demonstrated that (i) chloroplasts differentiate into chromoplasts during fruit coloring, and this differentiation is accompanied with a decrease in the chlorophyll/carotenoid ratio; (ii) BN-PAGE analysis reveals the profiles of macromolecular protein complexes among different types of plastids in citrus fruit; and (iii) the degradation rate of chlorophyll-protein complexes varies during the transition from chloroplasts to chromoplasts, with the stability generally following the order of LHCII > PS II core > LHC I > PS I core.ConclusionsOur optimized methods for both plastid separation and BN-PAGE assessment provide an opportunity for developing a better understanding of pigment-protein complexes and plastid transitions in plant fruit. These attempts also have the potential for expanding our knowledge on the sub-cellular level synchronism of protein changes and pigment metabolism during the transition from chloroplasts to chromoplasts.

Transcriptomic insights into the molecular mechanism of wheat response to stripe rust fungus

The wheat crop (Triticum aestivum L.) is the widely cultivated and most important staple foods of worlds. Stripe (yellow) rust is prompted by Puccinia striiformis f. sp. tritici (Pst) to reduces the yield and grain quality of the wheat significantly. Although many resistant cultivars have been successfully used in wheat breeding, the size of the regulating network and the underlying molecular mechanisms of wheat to response Pst still unknown. Therefore, in order to identify differentially expression genes (DEGs) and the regulate network related to Pst resistance, 15 cDNA libraries were constructed from wheat with CYR34 infection. In this study, a highly susceptible cv. Chuanyu12 (CY12) was used to study the transcriptome profiles after being inoculated with Pst physiological race CYR34. The DEGs were investigated at 24h, 48h, 72h, and 7 days post-inoculation. Certain key genes and pathways of response for Pst-CYR34 in CY12 were identified. The results revealed that Pst-CYR34 inhibited the DEGs related to energy metabolism, biosynthesis, carbon fixation, phenylalanine metabolism, and plant hormone signaling pathways after post-inoculation at 24h, 48h, 72h, and 7d. Light-harvesting chlorophyll protein complex in photosystem I and photosystem II; F-type ATPase, cytochrome b6/f/complex, and photosynthetic electron transport; ethylene, salicylic acid (SA), and jasmonic acid (JA); and lignin and flavonoids biosynthesis in CY12 are among the down-regulated DEGs. The expression patterns of these DEGs were verified via Quantitative Real-time PCR analysis. Our results give insights into the foundation for further exploring the molecular mechanisms regulating networks of Pst response and opens the door for bread wheat Pst resistance breeding.

Effects of Tetracycline on Scenedesmus obliquus Microalgae Photosynthetic Processes.

Tetracycline (TC) antibiotics can be detected worldwide in the aquatic environment due to their extensive use and low utilization efficiency, and they may affect the physiological processes of non-target organisms. In this study, the acute and sub-acute toxicities of TC on the freshwater microalga Scenedesmus obliquus were investigated with an emphasis on algal photosynthesis and transcription alterations during an 8 d TC exposure. The results showed that the IC10, IC30 and IC50 values were 1.8, 4.1 and 6.9 mg/L, respectively. During sub-acute exposure, the microalgae of the IC10 treatment was able to recover comparable growth to that of the control by day 7, while significantly lower cell densities were observed in the IC30 and IC50 treatments at the end of the exposure. The photosynthetic efficiency Fv/FM of S. obliquus first decreased as the TC concentration increased and then returned to a level close to that of the control on day 8, accompanied by an increase in photosynthetic activities, including light harvesting, electron transport and energy dissipation. Transcriptomic analysis of the IC10 treatment (1.8 mg/L TC) revealed that 2157 differentially expressed genes were up-regulated and 1629 were down-regulated compared with the control. KEGG and GO enrichments demonstrated that 28 photosynthesis-related genes involving light-harvesting chlorophyll protein complex, photosystem I, photosystem II, photosynthetic electron transport and enzymes were up-regulated, which may be the factor responsible for the enhanced photosynthesis and recovery of the microalgae. Our work may be helpful not only for gaining a better understanding of the environmental risk of TC at concentrations close to the real levels in natural waters, but also for explaining photosynthesis and related gene transcription induced by antibiotics.

Characterization, stability, and feasibility of long-term use of light-absorbing components of aqueous spinach extract-based photogalvanic electrolyte

In the present work, the photogalvanic cells have been studied with respect to the photo-stability and the long-term use of the electrolyte based on crude aqueous spinach extract sensitizer for solar energy harvesting. Further, the nature of chemical components present in the old and photo-decayed electrolyte and their current generation capacity has also not been investigated so far otherwise it is of much significance for durable use of the same electrolyte in cells. In earlier studies, the steady-state photo-generation of current for about two hours from crude spinach extract-based cell has been shown during illumination. But, the data for only two hours of the steady-state current generation is not sufficient to show the feasibility of working with photogalvanic cells. Therefore, to fill this research gap of lack of characterization of sensitizers’ molecules of crude spinach extract and lack of study on long-term use of this electrolyte (crude spinach extract-surfactant-reductant-alkali-water), the present extensive study has been done. The observed spectrum of crude spinach extract resembles that of chlorophyll–protein complex showing it is the main chemical component in extract absorbing light. A strong acid adversely affects the extract’s photogalvanics and high pH is friendly to the physiological and photogalvanic activity of the extract. The spectra of illuminated and very old crude spinach extract-NaOH-Sodium lauryl sulfate (NaLS)-Fructose photogalvanic electrolyte solution show negligible absorbance (540–700 nm) and zero absorbance (at 700 nm) suggesting the absence of chlorophyll due to its photo-degradation. When this photo-degraded electrolyte is again illuminated, the power output obtained is nearly equal to that for the first time illuminated fresh electrolyte. The observed current at zero time and after 2641 h from the same electrolyte used in long term is 50 mA cm−2 and 40 mA cm−2, respectively. It means that the fresh crude spinach extract, as well as the photo-degraded extract at high pH, are almost equally capable of power generation.

High-Light-Induced Degradation of Photosystem II Subunits’ Involvement in the Albino Phenotype in Tea Plants

The light-sensitive (LS) albino tea plant grows albinic shoots lacking chlorophylls (Chls) under high-light (HL) conditions, and the albinic shoots re-green under low light (LL) conditions. The albinic shoots contain a high level of amino acids and are preferential materials for processing quality green tea. The young plants of the albino tea cultivars are difficult to be cultivated owing to lacking Chls. The mechanisms of the tea leaf bleaching and re-greening are unknown. We detected the activity and composition of photosystem II (PSII) subunits in LS albino tea cultivar “Huangjinya” (HJY), with a normal green-leaf cultivar “Jinxuan” (JX) as control so as to find the relationship of PSII impairment to the albino phenotype in tea. The PSII of HJY is more vulnerable to HL-stress than JX. HL-induced degradation of PSII subunits CP43, CP47, PsbP, PsbR. and light-harvest chlorophyll–protein complexes led to the exposure and degradation of D1 and D2, in which partial fragments of the degraded subunits were crosslinked to form larger aggregates. Two copies of subunits PsbO, psbN, and Lhcb1 were expressed in response to HL stress. The cDNA sequencing of CP43 shows that there is no difference in sequences of PsbC cDNA and putative amino acids of CP43 between HJY and JX. The de novo synthesis and/or repair of PSII subunits is considered to be involved in the impairment of PSII complexes, and the latter played a predominant role in the albino phenotype in the LS albino tea plant.

Comparative analyses of nutritional strategies among the species within the genus Paragymnodinium (Gymnodiniales, Dinophyceae).

The genus Paragymnodinium exhibits various nutritional strategies despite the fact that each species has pigmented plastids. This provides an excellent opportunity to investigate the evolution of the mode of nutrition and the function of the plastids through comparative studies. In this study, we analyzed the growth, pigment composition, absorption spectra, variable chlorophyll a fluorescence, and photosynthetic carbon fixation capabilities of P. stigmaticum, P. asymmetricum, and P. inerme. The autotrophic species P. asymmetricum and P. inerme without resorting to any nutrition from prey organisms displayed high PSII activity and carbon fixation rates. The pigment compositions of these two species were identical to those of other typical peridinin-containing type dinoflagellates. On the contrary, the phagotrophic species P. stigmaticum showed heterotrophic growth, and the variable chlorophyll a fluorescence properties and carbon fixation rates indicated significantly lower photosynthetic competence relative to those of the above two species. Paragymnodinium stigmaticum also contained peridinin, but pigment content ratios of peridinin, chlorophyll c2, and β-carotene were significantly different from those of other two species. The absorption spectrum analysis revealed a redshift in the peak of the Qy band of chlorophyll a in P. stigmaticum, presumably due to a change in chlorophyll-protein complexes. We conclude that such distinct differences in nutritional strategies between members of the genus Paragymnodinium would provide a platform for the hypothetical loss of photosynthetic function leading to colorless dinoflagellates.

Structure-based model of fucoxanthin-chlorophyll protein complex: Calculations of chlorophyll electronic couplings.

Diatoms are a group of marine algae that are responsible for a significant part of global oxygen production. Adapted to life in an aqueous environment dominated by the blue-green light, their major light-harvesting antennae-fucoxanthin-chlorophyll protein complexes (FCPs)-exhibit different pigment compositions than of plants. Despite extensive experimental studies, until recently the theoretical description of excitation energy dynamics in these complexes was limited by the lack of high-resolution structural data. In this work, we use the recently resolved crystallographic information of the FCP complex from Phaeodactylum tricornutum diatom [Wang et al., Science 363, 6427 (2019)] and quantum chemistry-based calculations to evaluate the chlorophyll transition dipole moments, atomic transition charges from electrostatic potential, and the inter-chlorophyll couplings in this complex. The obtained structure-based excitonic couplings form the foundation for any modeling of stationary or time-resolved spectroscopic data. We also calculate the inter-pigment Förster energy transfer rates and identify two quickly equilibrating chlorophyll clusters.

Isolation of fucoxanthin chlorophyll protein complexes of the centric diatom Thalassiosira pseudonana associated with the xanthophyll cycle enzyme diadinoxanthin de-epoxidase.

In the present study, low concentrations of the very mild detergent n-dodecyl-α-d-maltoside in conjunction with sucrose gradient ultracentrifugation were used to prepare fucoxanthin chlorophyll protein (FCP) complexes of the centric diatom Thalassiosira pseudonana. Two main FCP fractions were observed in the sucrose gradients, one in the upper part and one at high sucrose concentrations in the lower part of the gradient. The first fraction was dominated by the 18 kDa FCP protein band in SDS-gels. Since this fraction also contained other protein bands, it was designated as fraction enriched in FCP-A complex. The second fraction contained mainly the 21 kDa FCP band, which is typical for the FCP-B complex. Determination of the lipid composition showed that both FCP fractions contained monogalactosyl diacylglycerol as the main lipid followed by the second galactolipid of the thylakoid membrane, namely digalactosyl diacylglycerol. The negatively charged lipids sulfoquinovosyl diacylglycerol and phosphatidyl glycerol were also present in both fractions in pronounced concentrations. With respect to the pigment composition, the fraction enriched in FCP-A contained a higher amount of the xanthophyll cycle pigments diadinoxanthin (DD) and diatoxanthin (Dt), whereas the FCP-B fraction was characterized by a lower ratio of xanthophyll cycle pigments to the light-harvesting pigment fucoxanthin. Protein analysis by mass spectrometry revealed that in both FCP fractions the xanthophyll cycle enzyme diadinoxanthin de-epoxidase (DDE) was present. In addition, the analysis showed an enrichment of DDE in the fraction enriched in FCP-A but only a very low amount of DDE in the FCP-B fraction. In-vitro de-epoxidation assays, employing the isolated FCP complexes, were characterized by an inefficient conversion of DD to Dt. However, in line with the heterogeneous DDE distribution, the fraction enriched in FCP-A showed a more pronounced DD de-epoxidation compared with the FCP-B.

STN7 Kinase Is Essential for Arabidopsis thaliana Fitness under Prolonged Darkness but Not under Dark-Chilling Conditions.

Reversible phosphorylation of photosystem II light harvesting complexes (LHCII) is a well-established protective mechanism enabling efficient response to changing light conditions. However, changes in LHCII phosphorylation were also observed in response to abiotic stress regardless of photoperiod. This study aimed to investigate the impact of dark-chilling on LHCII phosphorylation pattern in chilling-tolerant Arabidopsis thaliana and to check whether the disturbed LHCII phosphorylation process will impact the response of Arabidopsis to the dark-chilling conditions. We analyzed the pattern of LHCII phosphorylation, the organization of chlorophyll–protein complexes, and the level of chilling tolerance by combining biochemical and spectroscopy techniques under dark-chilling and dark conditions in Arabidopsis mutants with disrupted LHCII phosphorylation. Our results show that during dark-chilling, LHCII phosphorylation decreased in all examined plant lines and that no significant differences in dark-chilling response were registered in tested lines. Interestingly, after 24 h of darkness, a high increase in LHCII phosphorylation was observed, co-occurring with a significant FV/FM parameter decrease. The highest drop of FV/FM was detected in the stn7-1 line–mutant, where the LHCII is not phosphorylated, due to the lack of STN7 kinase. Our results imply that STN7 kinase activity is important for mitigating the adverse effects of prolonged darkness.

Chlorophyll f can replace chlorophyll a in the soluble antenna of dinoflagellates.

Chlorophyll f is a new type of chlorophyll isolated from cyanobacteria. The absorption and fluorescence characteristics of chlorophyll f permit these oxygenic-photosynthetic organisms to thrive in environments where white light is scarce but far-red light is abundant. To explore the ligand properties of chlorophyll f and its energy transfer profiles we established two different in vitro reconstitution systems. The reconstituted peridinin-chlorophyll f protein complex (chlorophyll f-PCP) showed a stoichiometry ratio of 4:1 between peridinin and chlorophyll f, consistent with the peridinin:chlorophyll a ratio from native PCP complexes. Using emission wavelength at 712nm, the excitation fluorescence featured a broad peak at 453nm and a shoulder at 511nm confirming energy transfer from peridinin to chlorophyll f. In addition, by using a synthetic peptide mimicking the first transmembrane helix of light-harvesting chlorophyll proteins of plants, we report that chlorophyll f, similarly to chlorophyll b, did not interact with the peptide contrarily to chlorophyll a, confirming the accessory role of chlorophyll f in photosystems. The binding of chlorophyll f, even in the presence of chlorophylls a and b, by PCP complexes shows the flexibility of chlorophyll-protein complexes and provides an opportunity for the introduction of new chlorophyll species to extend the photosynthetic spectral range.

Salt Stress Affects Plastid Ultrastructure and Photosynthetic Activity but Not the Essential Oil Composition in Spearmint (Mentha spicata L. var. crispa "Moroccan").

High levels of soil salinity affect plant growth, reproduction, water and ion uptake, and plant metabolism in a complex manner. In this work, the effect of salt stress on vegetative growth, photosynthetic activity, and chloroplast ultrastructure of spearmint (Mentha spicata L. var. crispa “Moroccan”) was investigated. After 2 weeks of low concentration treatments (5, 25, and 50 mM NaCl) of freshly cut shoots, we observed that the stem-derived adventitious root formation, which is a major mean for vegetative reproduction among mints, was completely inhibited at 50 mM NaCl concentration. One-week-long, high concentration (150 mM NaCl) salt stress, and isosmotic polyethylene glycol (PEG) 6000 treatments were compared in intact (rooted) plants and freshly cut, i.e., rootless shoots. Our data showed that roots have an important role in mitigating the deleterious effects of both the osmotic (PEG treatment) and specific ionic components of high salinity stress. At 50 mM NaCl or above, the ionic component of salt stress caused strong and irreversible physiological alterations. The effects include a decrease in relative water content, the maximal and actual quantum efficiency of photosystem II, relative chlorophyll content, as well as disorganization of the native chlorophyll-protein complexes as revealed by 77 K fluorescence spectroscopy. In addition, important ultrastructural damage was observed by transmission electron microscopy such as the swelling of the thylakoid lumen at 50 mM NaCl treatment. Interestingly, in almost fully dry leaf regions and leaves, granum structure was relatively well retained, however, their disorganization occurred in leaf chloroplasts of rooted spearmint treated with 150 mM NaCl. This loss of granum regularity was also confirmed in the leaves of these plants using small-angle neutron scattering measurements of intact leaves of 150 mM NaCl-stressed rooted plants. At the same time, solid-phase microextraction of spearmint leaves followed by gas chromatography and mass spectrometry (GC/MS) analyses revealed that the essential oil composition of spearmint was unaffected by the treatments applied in this work. Taken together, the used spearmint cultivar tolerates low salinity levels. However, at 50 mM NaCl concentration and above, the ionic components of the stress strongly inhibit adventitious root formation and thus their clonal propagation, and severely damage the photosynthetic apparatus.

Enhanced photosynthetic nitrogen use efficiency and increased nitrogen allocation to photosynthetic machinery under cotton domestication.

Domestication involves dramatic phenotypic and physiological diversifications due to successive selection by breeders toward high yield and quality. Although photosynthetic nitrogen use efficiency (PNUE) is a major trait for understanding leaf nitrogen economy, it is unclear whether PNUE of cotton has been improved under domestication. Here, we investigated the effect of domestication on nitrogen allocation to photosynthetic machinery and PNUE in 25 wild and 37 domesticated cotton genotypes. The results showed that domesticated genotypes had higher nitrogen content per mass (Nm), net photosynthesis under saturated light (Asat), and PNUE but similar nitrogen content per area (Na) compared with wild genotypes. As expected, in both genotypes, PNUE was positively related to Asat but negatively correlated with Na. However, the relative contribution of Asat to PNUE was greater than the contribution from Na. Domesticated genotypes had higher nitrogen allocation to light-harvesting (NL, nitrogen in light-harvesting chlorophyll-protein complex), to bioenergetics (Nb, total nitrogen of cytochrome f, ferredoxin NADP reductase, and the coupling factor), and to Rubisco (Nr) than wild genotypes; however, the two genotype groups did not differ in PNUEp, the ratio of Asat to Np (itself the sum of NL, Nb, and Nr). Our results suggest that more nitrogen allocation to photosynthetic machinery has boosted Asat under cotton domestication. Improving the efficiency of nitrogen use in photosynthetic machinery might be future aim to enhance Asat of cotton.

Enrichment of chlorophyll catabolic enzymes in grana margins and their cooperation in catabolic reactions

During leaf senescence, chlorophyll a and b are degraded through several enzymatic reactions, including chlorophyll b reductase, 7-hydroxymethyl chlorophyll a reductase, and Mg-dechelatase. Considering that the intermediates of the chlorophyll breakdown pathway are highly photoreactive, cooperative and efficient reactions of chlorophyll metabolic enzymes may protect chloroplasts from potential photo-oxidative damage. Here, we investigated the sub-organellar localization and cooperative reactions of the enzymes involved in the chlorophyll breakdown pathway by the fractionation of thylakoid membranes and enzymatic assays using recombinant proteins. We found that these enzymes were enriched in the grana margin fraction. Furthermore, we found that chlorophyll b reductase and Mg-dechelatase efficiently catabolized chlorophylls bound to the chlorophyll–protein complexes when these two enzymes were mixed. These results suggest that the co-localization of chlorophyll catabolic enzymes enables efficient chlorophyll breakdown. The results from this study highlight a key step forward in the investigation of the photosystem breakdown process.