Thylakoid Structure Research Articles

Response of photosynthetic capacity and antioxidative system of chloroplast in two wucai (Brassica campestris L.) genotypes against chilling stress.

Chilling stress during the growing season could cause a series of changes in wucai (Brassica campestris L.). WS-1 (chilling-tolerant genotype) and Ta2 (chilling-sensitive genotype) were sampled in present study to explore the chilling tolerance mechanisms. Our results indicated that photosynthetic parameters exhibited lower level in Ta2 than in WS-1 under chilling stress. The rapid chlorophyll fluorescence dynamics curve showed that chilling resulted in a greater inactivation of photosystem II reaction center in Ta2. Reactive oxygen species and malondialdehyde content of chloroplast in Ta2 were higher than WS-1. The ascorbate-glutathione cycle in chloroplast of WS-1 played a more crucial role than Ta2, which was confirmed by higher activities of antioxidant enzymes including Ascorbate peroxidase, Glutathione reductase, Monodehydroascorbate reductase and Dehydroascorbate reductase and higher content of AsA and GSH. In addition, the ultrastructure of chloroplasts in Ta2 was more severely damaged. After low temperature stress, the shape of starch granules in Ta2 changed from elliptical to round and the volume became larger than that of WS-1. The thylakoid structure of Ta2 also became dispersed from the original tight arrangement. Combined with our previous study under heat stress, WS-1 can tolerant both chilling stress and heat stress, which was partly due to a stable photosynthetic system and the higher active antioxidant system in plants, in comparison to Ta2.

Phenotype and TMT-based quantitative proteomics analysis of Brassica napus reveals new insight into chlorophyll synthesis and chloroplast structure

The conversion of light energy into chemical energy in leaves is very important for plant growth and development. During this process, chlorophylls and their derivatives are indispensable as their fundamental role in the energy absorption and transduction activities. Chlorophyll variation mutants are important materials for studying chlorophyll metabolism, chloroplast biogenesis, photosynthesis and related physiological processes. Here, a chlorophyll-reduced mutant (crm1) was isolated from ethyl methanesulfonate (EMS) mutagenized Brassica napus. Compared to wild type, crm1 showed yellow leaves, reduced chlorophyll content, fewer thylakoid stacks and retarded growth. Quantitative mass spectrometry analysis with Tandem Mass Tag (TMT) isobaric labeling showed that totally 4575 proteins were identified from the chloroplast of Brassica napus leaves, and 466 of which displayed differential accumulations between wild type and crm1. The differential abundance proteins were found to be involved in chlorophyll metabolism, photosynthesis, phagosome and proteasome. Our results suggest that the decreased abundance of chlorophyll biosynthetic enzymes, proteins involved in photosynthesis might account for the reduced chlorophyll content, impaired thylakoid structure, and reduction of plant productivity. The increased abundance of proteins involved in phagosome and proteasome pathways might allow plants to adapt the proteome to environmental conditions to ensure growth and survival due to chlorophyll reduction. Biological significancePhotosynthesis, which consists of light and dark reactions, is fundamental to biomass production. Chloroplast is regarded as the main site for photosynthesis. During photosynthesis, the pigment chlorophyll is essential for light harvesting and energy transfer. This work provides new insights into protein expression patterns, and enables the identification of many attractive candidates for investigation of chlorophyll biosynthesis, chloroplast structure and photosynthesis in Brassica napus. These findings may be applied to improve the photosynthetic efficiency by genetic engineering in crops.

Impaired Plastid Ribosomal Protein L3 Causes Albino Seedling Lethal Phenotype in Rice

The chloroplast is a semiautonomous photosynthetic organelle that is essential for plant growth and development, particularly in crops via manipulation of its photosynthetic capacity and the biosynthesis of carbon skeletons. Plastid ribosomal proteins (PRPs) are crucial for the establishment of the transcription/translation apparatus during chloroplast differentiation. In this study, we isolated and characterized T-DNA-tagged rice mutants with defective chloroplasts, named prpl3, that exhibited a distinct albino seedling lethality. Transmission electronic microscopy (TEM) observations showed that the grana stacks in the mutant were not properly formed, with disrupted thylakoid structures in their chloroplasts. Chlorophyll content was also significantly reduced in the leaves of prpl3 mutant seedlings. PRPL3 contains nuclear genes encoding PRPs localized to the chloroplasts, and prpl3 represents a novel mutant presentation of an impaired PRPL3 gene. Our findings also demonstrated that PRPL3 is responsible for phenotypic alterations by generating additional mutant alleles thereof using CRISPR/Cas9 systems. Expression levels of genes involved in photosynthesis and chloroplast development, including plastidial transcription and translation and photosynthesis, were altered in the prpl3 mutant. These results collectively demonstrate that nuclear-encoded PRPL3 is indispensable for the proper development of chloroplasts in rice.

GhYGL1d, a pentatricopeptide repeat protein, is required for chloroplast development in cotton

BackgroundThe pentatricopeptide repeat (PPR) gene family, which contains multiple 35-amino acid repeats, constitutes one of the largest gene families in plants. PPR proteins function in organelles to target specific transcripts and are involved in plant development and growth. However, the function of PPR proteins in cotton is still unknown.ResultsIn this study, we characterized a PPR gene YELLOW-GREEN LEAF (GhYGL1d) that is required for cotton plastid development. The GhYGL1d gene has a DYW domain in C-terminal and is highly express in leaves, localized to the chloroplast fractions. GhYGL1d share high amino acid-sequence homology with AtECB2. In atecb2 mutant, overexpression of GhYGL1d rescued the seedling lethal phenotype and restored the editing of accD and ndhF transcripts. Silencing of GhYGL1d led to the reduction of chlorophyll and phenotypically yellow-green leaves in cotton. Compared with wild type, GhYGL1d-silenced cotton showed significant deformations of thylakoid structures. Furthermore, the transcription levels of plastid-encoded polymerase (PEP) and nuclear-encoded polymerase (NEP) dependent genes were decreased in GhYGL1d-silenced cotton.ConclusionsOur data indicate that GhYGL1d not only contributes to the editing of accD and ndhF genes, but also affects the expression of NEP- and PEP-dependent genes to regulate the development of thylakoids, and therefore regulates leaf variegation in cotton.

Effects of exogenous putrescine on the ultrastructure of and calcium ion flow rate in lettuce leaf epidermal cells under drought stress

The alleviation effects of exogenous putrescine treatment on the ultrastructure of and calcium ion flow rate in lettuce (Lactuca sativa L.) leaf cells under drought stress were studied. Lettuce seedlings were treated with foliar sprays of 0.1 mM putrescine for 8 days, after which drought stress was simulated by using 10% polyethylene glycol 6000. The morphological characteristics of the seedlings and the calcium ion flow rate across stomatal guard cells were subsequently determined, and the leaf cell ultrastructure was observed via transmission electron microscopy. Under drought stress, the morphological characteristics of the seedlings decreased, and calcium ion influx was predominant in the guard cells. In addition, compared to that under control conditions, the stomatal density under drought stress conditions increased significantly, the open/closed stoma ratio was lower, and the degree of stomatal opening was smaller. Exogenous putrescine sprays effectively reduced the stomatal density, increased the degree of stomatal opening, and increased the proportion of open stomata. In addition, the chloroplasts became round in shape, the thylakoid structure became blurry in appearance, the number of starch grains decreased, many osmium granules were produced, and plasmolysis occurred in the mesophyll cells. However, the chloroplasts were elongated, the thylakoid structure was clear, the starch grains were abundant, few osmium granules were produced, and plasmolysis did not occur. The above results show that, by altering the leaf cell ultrastructure as well as the flow rate and direction of calcium ions in guard cells, exogenous putrescine effectively improves the drought tolerance of lettuce.

Physiological characterization and transcriptome analysis of a chlorosis mutant in pak choi

Leaf color mutants are not only ideal resources for studying the physiological mechanisms of plant photosynthesis, but they are also selection markers for hybrid production. In this study, the chlorosis mutant ‘564Y’ and its wild-type counterpart ‘564’ were used to explore physiological characteristics and gene expression in pak choi. Compared with those of wild type, the dry weight and fresh weight of ‘564Y’ were significantly reduced, but the hypocotyl length of ‘564Y’ was longer. Pigment analysis showed that the chlorophyll a and carotenoid contents of ‘564Y’ were lower than those of wild type. However, there was no significant difference in chlorophyll b content between these two varieties. The photosynthetic capacity of ‘564Y’ was lower than that of ‘564’, and the photosynthetic electron transfer ability and light energy conversion efficiency were weaker in the former. Transmission electron microscopy showed that the chloroplast structure of ‘564Y’ was not compact, the thylakoid structure was irregular, and the number of matrix, grana, and stacking layers was decreased. Transcriptome analysis revealed 2958 differentially expressed genes (DEGs), and 78 gene ontology terms and 35 Kyoto Encyclopedia of Genes and Genomes pathways were significantly enriched. Among the DEGs, nine genes were associated with the chlorophyll synthesis pathway (Bra031690, Bra012595, Bra005677, Bra022628, Bra026410, Bra032155, Bra039206, Bra029875, and Bra012511) and one gene was associated with photosynthesis (Bra038011). Among these genes, 3 were identified as related to auxin metabolism too (Bra032155, Bra039206, and Bra029875). Their differential expression in the ‘564Y’ mutant and wild-type plant was confirmed by qRT-PCR. These results lay the foundation for future genetic and functional genomic studies in pak choi.

Spatial organization of thylakoid network in higher plants

ABSTRACTAn intricate network of internal plastid membrane is formed in chloroplasts during plant ontogenesis. The network develops directly from a simple membrane system in proplastids or through a paracrystalline tubular prolamellar body (PLB) in etioplasts. A complex spatial structure of PLB interconnected with porous prothylakoids (PTs) is formed when the seedling growth proceeds without light. Upon illumination, PLB transforms through a multistage process into a lamellar system of stacked (grana) and loosely arranged (stroma) thylakoids. The chloroplast thylakoid network, the site of photochemical reactions, is one of the most important as well as most complicated membrane systems in nature. Thus, extensive studies on its possible spatial rearrangements are important for a better understanding of the crucial role of the thylakoid structure in the regulation of photosynthesis. Complementary structural methods are necessary to provide a complex image of spatial thylakoid arrangement. Therefore, in this review, we describe the results obtained by methods using both fixed and in vivo samples. The main goal of this review is to present the state of research on the three-dimensional (3D) structure of internal plastid membrane network, its spatial arrangement during plant ontogenesis and its modifications caused by different factors. Influence of different membrane components on the formation of the thylakoid network shape is also described. Additionally, we summarize recent 3D structural results in the field of thylakoid dynamics induced by environmental factors, especially of light-triggered changes related to rearrangements of the photosynthetic complexes.

Brassinosteroids Regulate Antioxidant System and Protect Chloroplast Ultrastructure of Autotoxicity-Stressed Cucumber (Cucumis sativus L.) Seedlings

Autotoxicity is a common problem being faced in protected vegetable cultivation system. Phytoremediation of plant autotoxicity is an emerging concept to minimize deterioration of soil environment and reduction of yield and quality of vegetable crops. Brassinosteroids (BRs) have been reported as a potential phytohormone to assist phytoremediation. However, the effects of BRs-induced autotoxicity stress on plant growth, photosynthesis and antioxidant defense system are poorly understood. Hence, we focused on the changes in physiological characteristics and ultrastructure of cucumber leaves in response to the application of 24-epibrassinolide (EBR) under autotoxicity stress conditions. The results showed that leaf area, plant height, fresh weight and dry weight of cucumber were obviously decreased under autotoxicity stress conditions. EBR application obviously improved the phenotypic characteristics of cucumber seedlings. Chlorophyll content, net photosynthetic rate, stomatal conductance and transpiration rate of cucumber leaves were markedly reduced under autotoxicity stress conditions. Application of EBR improved the photosynthetic pigments (chlorophyll a by 15.80%, chlorophyll b by 18.70% and total chlorophyll content by 17.30%), net photosynthetic rate by 36.40% and stomatal opening of leaves under autotoxicity stress conditions. EBR application also maintained the integrity of chloroplast and thylakoid structures under autotoxicity stress conditions. The activity of catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX) and antioxidative compounds ascorbate (AsA) and reduced glutathione (GSH) contents were markedly decreased, however, these were obviously increased after EBR application under autotoxicity stress. EBR application also increased the soluble sugar and protein, and proline concentration by 59.70%, 7.22% and 36.58%, respectively in the leaves of cucumber, decreased malondialdehyde by 24.13% and reactive oxygen species contents (H2O2 by 35.17%, O2− by 12.01% and •OH by 16.59%), and reduced the relative permeability of the cell membrane by 14.31%. These findings suggest that EBR application enhanced the photosynthetic capacity of leaves, maintained the integrity of chloroplast and thylakoid structures, and effectively alleviated the damage of membrane caused by lipid peroxidation and root damage under autotoxicity stress conditions. The growth inhibition effect of autotoxicity stress on cucumber was reduced by EBR application.

Subdiffraction-resolution live-cell imaging for visualizing thylakoid membranes.

The chloroplast is the chlorophyll-containing organelle that produces energy through photosynthesis. Within the chloroplast is an intricate network of thylakoid membranes containing photosynthetic membrane proteins that mediate electron transport and generate chemical energy. Historically, electron microscopy (EM) has been a powerful tool for visualizing the macromolecular structure and organization of thylakoid membranes. However, an understanding of thylakoid membrane dynamics remains elusive because EM requires fixation and sectioning. To improve our knowledge of thylakoid membrane dynamics we need to consider at least two issues: (i) the live-cell imaging conditions needed to visualize active processes invivo; and (ii) the spatial resolution required to differentiate the characteristics of thylakoid membranes. Here, we utilize three-dimensional structured illumination microscopy (3D-SIM) to explore the optimal imaging conditions for investigating the dynamics of thylakoid membranes in living plant and algal cells. We show that 3D-SIM is capable of examining broad characteristics of thylakoid structures in chloroplasts of the vascular plant Arabidopsis thaliana and distinguishing the structural differences between wild-type and mutant strains. Using 3D-SIM, we also visualize thylakoid organization in whole cells of the green alga Chlamydomonas reinhardtii. These data reveal that high light intensity changes thylakoid membrane structure in C.reinhardtii. Moreover, we observed the green alga Chromochloris zofingiensis and the moss Physcomitrella patens to show the applicability of 3D-SIM. This study demonstrates that 3D-SIM is a promising approach for studying the dynamics of thylakoid membranes in photoautotrophic organisms during photoacclimation processes.

Effect and mechanism of TiO2 nanoparticles on the photosynthesis of Chlorella pyrenoidosa

Titanium dioxide nanoparticles (n-TiO2) have been used in numerous applications, which results in their release into aquatic ecosystems and impact algal populations. A possible toxic mechanism of n-TiO2 on algae is via the disruption of the photosynthetic biochemical pathways, which yet remains to be demonstrated. In this study, Chlorella pyrenoidosa was exposed to different concentrations (0, 0.1, 1, 5, 10, and 20 mg/L) of a type of anatase n-TiO2, and the physiological, biochemical, and molecular responses involved in photosynthesis were investigated. The 96 h half growth inhibition concentration (IC50) of the n-TiO2 to algae was determined to be 9.1 mg/L. A variety of cellular and sub-cellular damages were observed, especially the blurry lamellar structure of thylakoids, indicating the n-TiO2 impaired the photosynthetic function of chloroplasts. Malondialdehyde (MDA) and glutathione disulfide (GSSG) significantly increased while the glutathione (GSH) content decreased. This implies the increased consumption of GSH by the increased intracellular oxidative stress upon n-TiO2 was insufficient to eliminate the lipid peroxidation. The contents of photosynthetic pigments, including chlorophyll a (Chl a) and phycobiliproteins (PBPs) in the exposed algal cells increased along with the up-regulation of genes encoding Chl a and photosystem II (PS II), which could be explained by a compensatory effect to overcome the toxicity induced by the n-TiO2. On the other hand, the photosynthetic activity was significantly inhibited, indicating the impairment on the photosynthesis via damaging the reaction center of PS II. In addition, lower productions of adenosine triphosphate (ATP) and glucose, together with the change of gene expressions suggested that the n-TiO2 disrupted the material and energy metabolisms in the photosynthesis. These findings support a paradigm shift of the toxic mechanism of n-TiO2 from physical and oxidative damages to metabolic disturbances, and emphasize the threat to the photosynthesis of algae in contaminated areas.

Effects of Cr2O3 nanoparticles on the chlorophyll fluorescence and chloroplast ultrastructure of soybean (Glycine max).

Chromic oxide nanoparticles (Cr2O3 NPs) are widely used in commercial factories and can cause serious environmental problems. However, the mechanism behind Cr2O3 NP-induced phytotoxicity remains unknown. In this study, the effects of Cr2O3 NPs on the growth, chlorophyll fluorescence, SEM-EDS analysis, and chloroplast ultrastructure of soybean (Glycine max) were investigated to evaluate its phytotoxicity. The growth of soybean treated with various Cr2O3 NP suspensions (0.01, 0.05, 0.1, and 0.5gL-1) was significantly inhibited. Specially, shoot and root biomass decreased by 9.9 and 46.3%, respectively. Besides, the maximum quantum yield of PSII (Fv/Fm) as well as the photochemical quenching (qP) decreased by 8-22 and 30-37%, respectively, indicating that the photosynthetic system was damaged when treated with Cr2O3 NPs. Moreover, the inhibition was confirmed by the reduction of Rubisco and MDH enzyme activity (by 54.5-86.4 and 26.7-96.5%, respectively). Overall, results indicated that the damage was caused by the destruction of chloroplast thylakoid structure, which subsequently reduced the photosynthetic rate. Our research suggests that Cr2O3 NPs can be transported and cause irreversible damage to soybean plants by inhibiting the activity of electron acceptors (NADP+) and destroying ultrastructure of chloroplasts, providing insights into plant toxicity issues. Graphical abstract ᅟ.

Effects of light intensities after anthesis on the photosynthetic characteristics and chloroplast ultrastructure in mesophyll cell of summer maize (Zea mays L. )

We examined the changes of photosynthetic characteristics and chloroplast ultrastructure in mesophyll cell of summer maize in response to different light intensities in the field, with the summer maize hybrid Denghai 605 as experimental material. Two treatments of both shading (S) and increasing light (L) from flowering to physiological maturity stage were designed, with the ambient sunlight treatment as control (CK). Under shading treatment, poorly developed thylakoid structure, blurry lamellar structure, loose granum, large gap between slices and warping granum were the major characteristics in chloroplast. Meanwhile, photosynthetic rate (Pn), transpiration rate, stomatal conductance, chlorophyll content, and actual photo-chemical efficiency (ΦPSII) decreased, whereas the maximal photochemical efficiency and non-photochemical quenching increased, which resulted in decreases in grain yield under shading treatment. However, a better development was observed in chloroplasts for L treatment, with the number of grana and lamellae increased and lamellae arranged compactly. In addition, Pn and ΦPSII increased under L treatment, which increased grain yield. The chloroplast arrangement dispersed in mesophyll cells and chloroplast ultrastructure was destroyed after shading, and then chlorophyll synthesis per unit leaf area and photosynthetic capacity decreased. In contrast, the number of grana and lamellae increased and lamellae arranged compactly after increasing light, which are beneficial for corn yield.

Arbuscular mycorrhizal fungi alleviate the heavy metal toxicity on sunflower (Helianthus annuus L.) plants cultivated on a heavily contaminated field soil at a WEEE-recycling site

An 8-week pot experiment was conducted to investigate the growth and responses of sunflower (Helianthus annuus L.) to arbuscular mycorrhizal (AM) fungal inoculations on a heavily heavy metal (HM)-contaminated (H) soil and a lightly HM-enriched (L) soil, both of which were collected from a waste electrical and electronic equipment (WEEE)-recycling site. Compared with the L soil, the H soil induced significantly larger (P<0.05) concentrations of Cd, Cu, Pb, Cr, Zn and Ni in sunflower (except for root Cr and shoot Ni), which impaired the thylakoid lamellar folds in leaves. The biomasses and P concentrations of shoots and roots, as well as the total P acquisitions per pot were all significantly decreased (P<0.05). Both Funneliformis mosseae (Fm) and F. caledonium (Fc) inoculation significantly increased (P<0.05) root mycorrhizal colonization. For the L soil, AM fungal inoculations had no significant effects on the soil-plant system, except for a decrease of soil pH and increases of soil available P and DTPA-extractable Zn concentrations with the Fm-inoculated treatment. For the H soil, however, AM fungal inoculations significantly increased (P<0.05) the biomasses and P concentrations of shoots and roots, as well as the total P acquisitions per pot, and significantly reduced (P<0.05) the concentrations of HMs in shoots (except for Cu and Pb with Fm- and Fc- inoculated treatments, respectively) and alleviated the toxicity symptoms of HMs in thylakoid structure of leaves. AM fungal inoculations in the H soil also significantly increased (P<0.05) the shoot uptake of HMs (except for Cr), and tended to decrease the total concentrations of HMs in soils. This suggests the potential application of AM fungi for both reducing HM stress and promoting phytoextraction of HM-contaminated soils caused by WEEE recycling.

Grana-Localized Proteins, RIQ1 and RIQ2, Affect the Organization of Light-Harvesting Complex II and Grana Stacking in Arabidopsis.

Grana are stacked thylakoid membrane structures in land plants that contain PSII and light-harvesting complex II proteins (LHCIIs). We isolated two Arabidopsis thaliana mutants, reduced induction of non-photochemical quenching1 (riq1) and riq2, in which stacking of grana was enhanced. The curvature thylakoid 1a (curt1a) mutant was previously shown to lack grana structure. In riq1 curt1a, the grana were enlarged with more stacking, and in riq2 curt1a, the thylakoids were abnormally stacked and aggregated. Despite having different phenotypes in thylakoid structure, riq1, riq2, and curt1a showed a similar defect in the level of nonphotochemical quenching of chlorophyll fluorescence (NPQ). In riq curt1a double mutants, NPQ induction was more severely affected than in either single mutant. In riq mutants, state transitions were inhibited and the PSII antennae were smaller than in wild-type plants. The riq defects did not affect NPQ induction in the chlorophyll b-less mutant. RIQ1 and RIQ2 are paralogous and encode uncharacterized grana thylakoid proteins, but despite the high level of identity of the sequence, the functions of RIQ1 and RIQ2 were not redundant. RIQ1 is required for RIQ2 accumulation, and the wild-type level of RIQ2 did not complement the NPQ and thylakoid phenotypes in riq1 We propose that RIQ proteins link the grana structure and organization of LHCIIs.

Leaf senescence: an overview

Senescence constitutes the last phase of plant growth and is characterized by a series of degenerative events that decrease metabolic activities and cause the death of cells, tissues and organs. Yellowing of leaves is a morphological indicator of senescence, which results due to loss of green pigments, i.e., chlorophylls. Alteration in mitochondrial and thylakoid structure, shrinkage of chloroplast and nucleus are prominent effects observed during senescence. Essential macromolecules like carbohydrates, proteins, lipids, nucleic acids, and photosynthetic pigments such as chlorophyll and carotenoids are degraded during senescence and these degraded constituents play important role in nutrient recycling mechanisms, which are mediated by senescence associated genes (SAGs). Sugar senescing mechanism, down regulation of photosynthetic genes, phytohormones, reactive oxygen species and SAGs play important role in signal transduction pathways to initiate progress and terminate the senescence process. The levels of heavy metals have increased in the environment due to various anthropogenic activities, which produce phytotoxic effects and reduce productivities. This review paper describes various biochemical/metabolic changes associated with leaf senescence.

Effects of shading on the photosynthetic characteristics and mesophyll cell ultrastructure of summer maize.

A field experiment was conducted to study the effects of shading on the photosynthetic characteristics and mesophyll cell ultrastructure of two summer maize hybrids Denghai605 (DH605) and Zhengdan958 (ZD958). The ambient sunlight treatment was used as control (CK) and shading treatments (40% of ambient sunlight) were applied at different growth stages from silking (R1) to physiological maturity (R6) (S1), from the sixth leaf stage (V6) to R1 (S2), and from seeding to R6 (S3), respectively. The net photosynthetic rate (P n) was significantly decreased after shading. The greatest reduction of P n was found at S3 treatment, followed by S1 and S2 treatments. P n of S3 was decreased by 59 and 48% for DH605, and 39 and 43% for ZD958 at tasseling and milk-ripe stages, respectively, compared to that of CK. Additionally, leaf area index (LAI) and chlorophyll content decreased after shading. In terms of mesophyll cell ultrastructure, chloroplast configuration of mesophyll cells dispersed, and part of chloroplast swelled and became circular. Meanwhile, the major characteristics of chloroplasts showed poorly developed thylakoid structure at the early growth stage, blurry lamellar structure, loose grana, and a large gap between slices and warping granum. Then, plasmolysis occurred in mesophyll cells and the endomembrane system was destroyed, which resulted in the dissolution of cell membrane, karyotheca, mitochondria, and some membrane structures. The damaged mesophyll cell ultrastructure led to the decrease of photosynthetic capacity, and thus resulted in significant yield reduction by 45, 11, and 84% in S1, S2, and S3 treatments, respectively, compared to that of CK.

Live-cell visualization of excitation energy dynamics in chloroplast thylakoid structures.

The intricate molecular processes underlying photosynthesis have long been studied using various analytic approaches. However, the three-dimensional (3D) dynamics of such photosynthetic processes remain unexplored due to technological limitations related to investigating intraorganellar mechanisms in vivo. By developing a system for high-speed 3D laser scanning confocal microscopy combined with high-sensitivity multiple-channel detection, we visualized excitation energy dynamics in thylakoid structures within chloroplasts of live Physcomitrella patens cells. Two distinct thylakoid structures in the chloroplast, namely the grana and stroma lamellae, were visualized three-dimensionally in live cells. The simultaneous detection of the shorter (than ~670 nm) and longer (than ~680 nm) wavelength regions of chlorophyll (Chl) fluorescence reveals different spatial characteristics—irregular and vertical structures, respectively. Spectroscopic analyses showed that the shorter and longer wavelength regions of Chl fluorescence are affected more by free light-harvesting antenna proteins and photosystem II supercomplexes, respectively. The high-speed 3D time-lapse imaging of the shorter and longer wavelength regions also reveals different structural dynamics—rapid and slow movements within 1.5 seconds, respectively. Such structural dynamics of the two wavelength regions of Chl fluorescence would indicate excitation energy dynamics between light-harvesting antenna proteins and photosystems, reflecting the energetically active nature of photosynthetic proteins in thylakoid membranes.

The differences between two tea varieties in their response to natural cold conditions

ABSTRACTIn this study, two tea varieties (Camellia sinensis cv. CsCr02 and CsCr03) were studied under natural cold conditions to investigate the differences of thylakoid membrane structure, fatty acid desaturase (FAD) genes, and fatty acid (FA) composition in response to low temperatures. Quantitative real-time polymerase chain reaction assay confirmed that the expression of CsFAD8 in leaves of the two tea varieties was much higher than that of CsFAD7. The expression of both genes in CsCr03 was higher than in CsCr02. Electron microscopy showed that the damage to the thylakoid membrane in CsCr03 was less severe than in CsCr02 during the decrease of natural temperature. Changes in thylakoid membrane integrity coincided with the gradual decline in chlorophyll concentration and chlorosis of the tea leaves. With the decrease of natural temperature, a decline in the trienoic FA species (16:3 and 18:3) and an increase in the dienoic species (18:2) were observed in the two varieties. These results indicate that CsCr03 has higher cold adaptability compared to CsCr02.

Different response of photosystem II to short and long-term drought stress in Arabidopsis thaliana.

Short- and long-term drought stress on photosystem II (PSII) and oxidative stress were studied in Arabidopsis thaliana. Under drought stress, chlorophyll (Chl) content, Chl fluorescence, relative water content and oxygen evolution capacity gradually decreased, and the thylakoid structure was gradually damaged. Short-term drought stress caused a rapid disassembly of the light-harvesting complex II (LHCII). However, PSII dimers kept stable under the short-term drought stress and significantly decreased only after 15 days of drought stress. Immunoblotting analysis of the thylakoid membrane proteins showed that most of the photosystem proteins decreased after the stress, especially for Lhcb5, Lhcb6 and PsbQ proteins. However, surprisingly, PsbS significantly increased after the long-term drought stress, which is consistent with the substantially increased non-photochemical quenching (NPQ) after the stress. Our results suggest that the PSII-LHCII supercomplexes and LHCII assemblies play an important role in preventing photo-damages to PSII under drought stress.

Albino Leaf1 That Encodes the Sole Octotricopeptide Repeat Protein Is Responsible for Chloroplast Development.

Chloroplast, the photosynthetic organelle in plants, plays a crucial role in plant development and growth through manipulating the capacity of photosynthesis. However, the regulatory mechanism of chloroplast development still remains elusive. Here, we characterized a mutant with defective chloroplasts in rice (Oryza sativa), termed albino leaf1 (al1), which exhibits a distinct albino phenotype in leaves, eventually leading to al1 seedling lethality. Electronic microscopy observation demonstrated that the number of thylakoids was reduced and the structure of thylakoids was disrupted in the al1 mutant during rice development, which eventually led to the breakdown of chloroplast. Molecular cloning revealed that AL1 encodes the sole octotricopeptide repeat protein (RAP) in rice. Genetic complementation of Arabidopsis (Arabidopsis thaliana) rap mutants indicated that the AL1 protein is a functional RAP. Further analysis illustrated that three transcript variants were present in the AL1 gene, and the altered splices occurred at the 3' untranslated region of the AL1 transcript. In addition, our results also indicate that disruption of the AL1 gene results in an altered expression of chloroplast-associated genes. Consistently, proteomic analysis demonstrated that the abundance of photosynthesis-associated proteins is altered significantly, as is that of a group of metabolism-associated proteins. More specifically, we found that the loss of AL1 resulted in altered abundances of ribosomal proteins, suggesting that RAP likely also regulates the homeostasis of ribosomal proteins in rice in addition to the ribosomal RNA. Taken together, we propose that AL1, particularly the AL1a and AL1c isoforms, plays an essential role in chloroplast development in rice.