Chloroplast Ultrastructure Research Articles

Unraveling subcellular functional traits: Adaptive insights into chloroplast ultrastructure in nonmodel species.

This essay discusses how the ultrastructural changes in chloroplasts, particularly the mechanisms of thylakoid membrane unstacking, help maintain the photosynthetic performance of photosystem II (PSII) under stress conditions. This phenomenon may facilitate the repair of damaged PSII by providing access to the repair machinery. It is argued that this PSII repair mechanism accelerates PSII recovery, optimizing photosynthetic processes in stressed plants. Although some studies demonstrate the relationship between thylakoid membrane unstacking in stress conditions, these studies were developed with model species under controlled conditions. Thus, this essay serves as a validation tool for these previous studies, because it demonstrates that the relationships between ultrastructural changes in chloroplasts and the functioning of PSII are essential acclimative strategies for nonmodel plants to survive the constant edaphoclimatic changes of natural environments. Understanding these subcellular dynamics can significantly inform biologists about the plastic potential of plants, especially in heterogeneous environments. An integrated approach in future studies is necessary, highlighting the importance of exploring plant functional traits at multiple scales, because subcellular characteristics have great potential to understand plant acclimatization.

Aluminium bioavailability and toxicity disrupted chloroplast structure and inhibited inorganic carbon utilization and nutrient uptake in Vallisneria natans at acidic and alkaline pH

Vallisneria natans, as submerged aquatic plants, face significant threats from aluminium (Al) toxicity. While the effects of Al at low pH on terrestrial plants have been extensively studied, there is a lack of research on the impacts of both low and high pH on chloroplast ultrastructure and nutrient uptake in submerged plants. This research is important as it aims to fill this gap by exposing the leaves of Vallisneria natans to 100 μM Al at varying pH levels (4.5, 5.5, 7.5, and 9.5) for 48 hours. The results showed that inorganic carbon (CT), CO2, and HCO3 content increased at extreme pH levels (4.5 and 9.5), suggesting decreased inorganic carbon utilization under Al stress. Additionally, photosystem II efficiency and electron transport rate were significantly reduced at extreme pH levels, highlighting the sensitivity of V. natans to Al. Chlorophyll a and total chlorophyll content were significantly lower at pH 4.5 compared to pH 7.5. Chloroplast structural disruptions were evident at extreme pH levels coupled with Al exposure, whereas minimal injury was observed at pH 5.5 and 7.5. The study also noted vacuole enlargement, altered plasma membrane permeability, and hematoxylin staining, indicating Al accumulation in leaves. ICP analysis revealed increased Al content at extreme pH levels, underscoring heightened Al bioavailability and toxicity. Significant reductions in macro and micronutrient content (P, Mg, K, Fe, Zn, B, Mn) were observed, likely due to Al-induced root and cell damage and altered nutrient uptake. These findings emphasize the complex interplay between Al exposure, pH fluctuations, and their cascading effects on the physiology and elemental composition of Vallisneria natans, highlighting the need for further research and environmental management strategies.

Long-term effects of silver nanoparticles and mineral nutrition components on the photosynthetic processes, chloroplast ultrastructure and productivity of Solanum lycopersicum plants

The effects of silver nanoparticles (AgNPs), both alone and in combination with mineral nutrients, on the growth and photosynthesis of Solanum lycopersicum plants during ontogeny were studied. The experiment involved weekly applications of 10 μmol of AgNPs for 15 weeks in a greenhouse over a summer period. A comprehensive characterization of the AgNPs was performed via TEM, ESI/EELS, and zeta potential measurements before and throughout the experiment. The activity of PSII, stomatal conductivity, photosynthesis, transpiration and respiration rates were measured, and the photosynthetic pigments, chloroplast ultrastructure, and dry and fresh masses of leaves, roots, and fruits were assessed. The results indicated that combining AgNPs with mineral nutrients increased PSII activity and the photosynthesis rate and altered the chloroplast ultrastructure. However, the use of mineral nutrients or AgNPs alone did not induce these changes. Atomic absorption spectrometry detected AgNPs in all the plant organs except the fruits. The highest fruit yield was associated with Veni Prisma®, a commercial product containing colloidal silver, which also caused desynchronized fruit maturation. This study hypothesizes that the synergistic effect of AgNPs and mineral nutrients enhances silver accumulation in chloroplasts, improving light utilization and photosynthetic efficiency, particularly under low light, thus increasing fruit quantity and dry mass. Conversely, long-term use of AgNPs alone was accompanied by silver accumulation outside the chloroplasts and did not lead to increased photosynthesis or an increase in fresh fruit mass.

Agrobacterium-mediated transformation of B. juncea reveals that BjuLKP2 functions in plant yellowing.

A stable Agrobacterium-mediated transformation system was constructed for B. juncea, and BjuLKP2 was overexpressed, leading to plant yellowing. A stable and efficient transformation system is necessary to verify gene functions in plants. To establish an Agrobacterium-mediated transformation system for B. juncea, various factors, including the explant types, hormone combination and concentration, infection time and concentration, were optimized. Eventually, a reliable system was established, and two BjuLKP2 overexpression (OE) lines, which displayed yellowing of cotyledons, shoot tips, leaves and flower buds, as well as a decrease in total chlorophyll content, were generated. qRT-PCR assays revealed significant upregulation of five chlorophyll synthesis genes and downregulation of one gene in the BjuLKP2 OE line. Furthermore, antioxidant capacity assays revealed reduced activities of APX, CAT and SOD, while POD activity increased in the BjuLKP2 OE26. Additionally, the kinetic determination of chlorophyll fluorescence induction suggested a decrease in the photosynthetic ability of BjuLKP2 OE26. GUS assays revealed the expression of BjuLKP2 in various tissues, including the roots, hypocotyls, cotyledons, leaf vasculature, trichomes, sepals, petals, filaments, styles and stigma bases, but not in seeds. Scanning electron revealed alterations in chloroplast ultrastructure in both the sponge and palisade tissue. Collectively, these findings indicate that BjuLKP2 plays a role in plant yellowing through a reduction in chlorophyll content and changes in chloroplasts structure.

Exogenous melatonin delays yellowing in harvested broccoli by maintaining chloroplast integrity

Yellowing of broccoli is a crucial limiting factor for its commercial value and consumer acceptance during postharvest. In this study, the impacts of exogenous melatonin (MEL) on chlorophyll content and fluorescence, as well as ultrastructure and membrane lipid metabolism of chloroplasts in broccoli were investigated during postharvest. The results showed that MEL treatment (200 μmol L−1) maintained the chlorophyll content, chloroplast autofluorescence and integral structure, and reduced the level ofserotonin in the chloroplasts in broccoli. Also, MEL treatment inhibited the membrane lipid peroxidation of chloroplasts, as indicated by low levels of superoxide anion (O2-), hydrogen peroxide (H2O2) and malondialdehyde (MDA), and high levels of endogenous MEL. In addition, the stability and fluidity of chloroplast membranes were also better maintained in the treated broccoli via increasing the contents of phosphatidylglyceroland (PG), monogalactosyldiglyceride (MGDG), digalactosyldiglyceride (DGDG) and unsaturated fatty acids as well as decreasing saturated fatty acid content and the activities of lipoxygenase (LOX) and lipase (LPS). Thus, the application of MEL facilitated the maintenance of chloroplast integrity, thus contributing to yellowing postponement and the extension of the storage life of broccoli.

Alteration in lipid metabolism is involved in nitrogen deficiency response in wheat seedlings

Changes of membrane lipid composition contribute to plant adaptation to various abiotic stresses. Here, a comparative study was undertaken to investigate the mechanisms of how lipid alteration affects plant growth and development under nitrogen (N) deficiency. Two wheat cultivars: the N deficiency-tolerant cultivar Xiaoyan 6 (XY) and the N deficiency-sensitive cultivar Aikang 58 (AK) were used to test if the high N-deficiency tolerance was related with lipid metabolism. The results showed that N deficiency inhibited the morpho-physiological parameters in both XY and AK cultivars, which showed a significant decrease in biomass, N content, photosynthetic efficiency, and lipid contents. However, these decreases were more pronounced in AK than XY. In addition, XY showed a notable increase in fatty acid unsaturation, relatively well-maintained chloroplast ultrastructure, and minimized damage of lipid peroxidation and enhanced PSII activity under N-deficient condition, as compared with AK. Transcription levels of many genes involved in lipid biosynthesis and fatty acid desaturation were up-regulated in response to N deficiency in two wheat cultivars, while the expressions were much higher in XY than AK under N deficiency. These results highlight the importance of alterations in lipid metabolism in N deficiency tolerance in wheat. High levels of lipid content and unsaturated fatty acids maintained the membrane structure and function, contributing to high photosynthesis and antioxidant capacities, thereby improved the tolerance to N deficiency.

2,2',4,4'-Tetrabromodiphenyl Ether (BDE-47) at Environmental Levels Influenced Photosynthesis in the Mangrove Species Kandelia obovata.

2,2',4,4'-tetra-bromodiphenytol ether (BDE-47) is one of the ubiquitous organic pollutants in mangrove sediments. To reveal the toxic effects of BDE-47 on mangrove plants, the mangrove species Kandelia obovate was used to investigate the photosynthetic capacity effects and the molecular mechanisms involved after BDE-47 exposure at environment-related levels (50, 500, and 5000 ng g-1 dw). After a 60-day exposure, the photosynthetic capacity was inhibited in K. obovata seedlings, and a decrease in the stomatal density and damage in the chloroplast ultrastructure in the leaves were found. Transcriptome sequencing showed that, following exposure to BDE-47, gene expression in photosynthesis-related pathways was predominantly suppressed in the leaves. The bioinformatics analysis indicated that BDE-47 exerts toxicity by inhibiting photosystem I activity and chlorophyll a/b-binding protein-related genes in the leaves of K. obovata. Thus, this study provides preliminary theoretical evidence for the toxic mechanism effect of BDE-47 on photosynthesis in mangrove species.

Response of selenium pools to drought stress by regulating physio‑biochemical attributes and anatomical changes in Gentiana macrophylla

Selenium (Se), as a vital stress ameliorant, possesses a beneficial effect on mediating detrimental effects of environmental threats. However, the mechanisms of Se in mitigating the deleterious effects of drought are still poorly understood. Gentiana macrophylla Pall. is a well-known Chinese medicinal herb, and its root, as the main medicinal site, has significant therapeutic effects. The purpose of this experiment was to investigate the functions of Se on the seedling growth and physiobiochemical characteristics in G. macrophylla subjected to drought stress. The changes in microstructure and chloroplast ultrastructure of G. macrophylla leaves under drought exposure were characterized by scanning electron microscopy (SEM), scanning electron microscopes and energy dispersive X-Ray spectroscope (SEM-EDX), and transmission electron microscopy (TEM), respectively. Results revealed that drought stress induced a notable increase in oxidative toxicity in G. macrophylla, as evidenced by elevated levels of hydrogen peroxide (H2O2), lipid peroxidation (MDA), enhanced antioxidative response, decreased plant photosynthetic function, and inhibited plant growth. Chloroplasts integrity with damaged membranes and excess osmiophilic granule were observed in the drought-stressed plants. Se supplementation notably recovered the stomatal morphology, anatomical structure damage, and chloroplast ultrastructure of G. macrophylla leaves caused by drought exposure. Exogenous Se application markedly enhanced SPAD, photosynthetic stomatal exchange parameters, and photosystem II activity. Se supplementation significantly promoted the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT), while reducing levels of MDA, superoxide anion (O2–.) and H2O2, and improving membrane integrity. Furthermore, the ameliorative effects of Se were also suggested by increased contents of osmotic substances (soluble sugar and proline), boosted content of gentiopicroside and loganinic acid in roots, and alleviated the inhibition in plant growth and biomass. Fourier transform infrared (FTIR) analysis of Se-treated G. macrophylla roots under drought stress demonstrated that Se-stimulated metabolites including O-H, C-H, N-H, C-N, and CO functional groups, were involved in resisting drought stress. Correlation analysis indicated an obvious negative correlation between growth parameters and MDA, O2–. and H2O2 content, while a positive correlation with photosynthetic gas exchange parameters. Principal component analysis (PCA) results explained the total variance into two principal components contributing the maximum (93.50 %) among the drought exposure with or without Se due to the various experiment indexes. In conclusion, Se exerts beneficial properties on drought-induced detrimental effects in G. macrophylla by relieving oxidative stress, improving photosynthesis indexes, PSII activity, regulating anatomical changes, altering levels of gentiopicroside and loganinic acid, and promoting growth of drought-stressed G. macrophylla.

Different Oligosaccharides Induce Coordination and Promotion of Root Growth and Leaf Senescence during Strawberry and Cucumber Growth

Oligosaccharides, as a wide type of polysaccharide, have a broad antimicrobial spectrum and promote development as plant growth stimulants. To investigate the regulation effects of different oligosaccharides on the dynamic changes of chlorophyll content, leaf fluorescence, root activity and morphology, and chloroplast ultrastructure, as well as the yields and yield components of strawberry and cucumber, typical greenhouse experiments were conducted over two years (2021–2022). The experimental plants were foliar sprayed with tap water (CK), chitosan oligosaccharide (CSOS), and mixed oligosaccharides (MixOS) five times before flowering. The conventional management (CM) was conducted as a conventional control. The findings of the present study suggest that the application of MixOS has the greatest regulation effects on delayed leaf senescence, well-developed roots, and higher fruit productions of strawberry and cucumber. Exogenous MixOS resulted in significant increases in SPAD values, maximum photochemical efficiency (Fv/Fm), and photochemical quenching coefficiency (qP); they were increased by 1.94–28.96%, 5.41–33.89%, and 9.93–62.07%, compared to the CSOS, CM, and CK treatments, respectively. The orderly and steady structure of thylakoids in the chloroplast, and the randomly distributed starch grains, could be clearly observed in the MixOS treatment, while the non-photochemical quenching (NPQ) was correspondingly reduced by 19.04–45.92%. Meanwhile, the remarkable promotion of root activity and root surface morphology indicators (i.e., root length, surface area, average diameter, and volume) could be observed when exposed to the MixOS treatments, and the total yields of strawberry and cucumber were all increased by 12.40–25.57%. These findings suggest that the mixed oligosaccharides mainly promote the coordinated growth of root and shoot, which leads to the improved yields of strawberry and cucumber.

Understanding the interplay between strigolactone and nitric oxide in alleviating cadmium-induced toxicity in Artemisia annua

Metal stress severely limits the development and productivity of medicinal plants like A. annua. Strigolactone (SL), a relatively new plant hormone, and nitric oxide (NO) have demonstrated potential in mitigating HM-related symptoms in plants. Both SLs and NO are regulatory signals that play different roles under stressful conditions. At the moment, it is unclear how SLs and NO interact in plants under cadmium (Cd) stress. In this study, Artemisia annua was utilized to study the role and interaction of SL and NO for Cd stress tolerance. To investigate this, we undertook a detailed physiological, anatomical, and histochemical examination to determine the mechanisms by which SLs and NO reduce the effects of Cd stress on A. annua plants. Cadmium exposure hindered the growth of A. annua plants, influenced several chlorophyll fluorescence attributes, altered antioxidant enzyme activity, lowered cell viability, and caused lipid peroxidation, oxidative damage, and chloroplast ultrastructure abnormalities. A. annua plants grew better when treated with 4 μM synthetic SL analogue GR24 and 200 μM NO donor sodium nitroprusside. Meanwhile, under Cd stress, GR24 and/or SNP spray effectively boosted chlorophyll and carotenoid content, increased antioxidant enzyme activity, and elevated proline and glutathione levels. Treatment with SNP and/or GR24 improved chloroplast ultrastructure and chlorophyll fluorescence characteristics in plants under Cd-stress, resulting in better photosynthetic performance. It also reduced Cd accumulation, increased membrane permeability, and regulated stomatal behavior, resulting in enhanced stomatal conductance in Cd-stressed plants. The application of these growth regulators to plants under Cd-stress mitigated the decrease in glandular trichome dimensions and artemisinin content as observed from the HPLC results. Our results indicate that GR24 and NO, either individually or in combination, can be very effective in reducing Cd-induced damage in A. annua.

Differential Transcription Profiling Reveals the MicroRNAs Involved in Alleviating Damage to Photosynthesis under Drought Stress during the Grain Filling Stage in Wheat.

To explore the possible novel microRNA (miRNA) regulatory pathways in Zhengmai 1860, a newly cultivated drought-tolerant wheat (Triticum aestivum L.) cultivar, miRNA transcriptome sequencing of the flag leaves of Zhengmai 1860, drought-sensitive variety Zhoumai 18, and drought-resistant variety Bainong 207 was performed during the grain filling stage. We also observed changes in the chloroplast ultrastructure, phytohormone levels, and antioxidant- and photosynthesis-related physiological indicators in three wheat varieties. The results showed that the flag leaves of the drought-tolerant variety Zhengmai 1860 had higher chlorophyll contents and net photosynthetic rates than those of Zhoumai 18 under drought stress during the grain filling stage; in addition, the chloroplast structure was more complete. However, there was no significant difference between Zhengmai 1860 and Bainong 207. MiRNA transcriptome analysis revealed that the differential expression of the miRNAs and mRNAs exhibited variable specificity. The KEGG pathway enrichment results indicated that most of the genes were enriched in the MAPK signaling pathway, plant hormone signal transduction, photosynthetic antennae protein, and amino acid and carbohydrate metabolism. In the drought-tolerant cultivar Zhengmai 1860, tae-miR408 was targeted to regulate the allene oxide synthase (AOS) gene, inhibit its expression, reduce the AOS content, and decrease the synthesis of jasmonic acid (JA) and abscisic acid (ABA). The results of this study suggest that Zhengmai 1860 could improve the photosynthetic performance of flag leaves by inhibiting the expression of genes involved in the JA pathway through miRNAs under drought conditions. Moreover, multiple miRNAs may target chlorophyll, antioxidant enzymes, phytohormone signal transduction, and other related pathways; thus, it is possible to provide a more theoretical basis for wheat molecular breeding.

Alterations in the Anatomy and Ultrastructure of Leaf Blade in Norway Maple (Acer platanoides L.) Growing on Mining Sludge: Prospects of Using This Tree Species for Phytoremediation.

Alterations in leaf architecture can be used as an indicator of the substrate toxicity level as well as the potential of a given plant species in the phytoremediation of polluted areas, e.g., mining sludge. In this work, we demonstrated, for the first time, the nature and scale of alterations in leaf architecture at the tissue and cellular levels occurring in Norway maple growing on mining sludge originating from a copper mine in Lubin (Poland). The substrate differs from other mine wastes, e.g., calamine or serpentine soils, due to an extremely high level of arsenic (As). Alterations in leaf anatomy predominantly included the following: (1) a significant increase in upper epidermis thickness; (2) a significant decrease in palisade parenchyma width; (3) more compact leaf tissue organization; (4) the occurrence of two to three cell layers in palisade parenchyma in contrast to one in the control; (5) a significantly smaller size of cells building palisade parenchyma. At the cellular level, the alterations included mainly the occurrence of local cell wall thickenings-predominantly in the upper and lower epidermis-and the symptoms of accelerated leaf senescence. Nevertheless, many chloroplasts showed almost intact chloroplast ultrastructure. Modifications in leaf anatomy could be a symptom of alterations in morphogenesis but may also be related to plant adaptation to water deficit stress. The occurrence of local cell wall thickenings can be considered as a symptom of a defence strategy involved in the enlargement of apoplast volume for toxic elements (TE) sequestration and the alleviation of oxidative stress. Importantly, the ultrastructure of leaf cells was not markedly disturbed. The results suggested that Norway maple may have good phytoremediation potential. However, the general shape of the plant, the significantly smaller size of leaves, and accelerated senescence indicated the high toxicity of the mining sludge used in this experiment. Hence, the phytoremediation of such a substrate, specifically including use of Norway maple, should be preceded by some amendments-which are highly recommended.

Physiological and Structural Changes in Leaves of Platycrater arguta Seedlings Exposed to Increasing Light Intensities.

Understanding the light adaptation of plants is critical for conservation. Platycrater arguta, an endangered deciduous shrub endemic to East Asia, possesses high ornamental and phylogeographic value. However, the weak environmental adaptability of P. arguta species has limited its general growth and conservation. To obtain a deeper understanding of the P. arguta growth conditions, we examined the leaf morphology and physiology via anatomical and chloroplast ultrastructural analyses following exposure to different natural light intensities (full light, 40%, and 10%). The findings indicated that P. arguta seedings in the 10% light intensity had significantly improved leaf morphological characteristics and specific leaf area compared to those exposed to other intensities. The net photosynthetic rate, chlorophyll (Chl) content, photosynthetic nitrogen use efficiency (PNUE), and photosynthetic phosphorus use efficiency (PPUE) exhibited marked increases at a 10% light intensity compared to both 40% light and full light intensities, whereas the light compensation point and dark respiration levels reached their lowest values under the 10% light condition. With reduced light, leaf thickness, palisade tissue, spongy tissue, and stomatal density significantly decreased, whereas the stomatal length, stomatal width, and stomatal aperture were significantly elevated. When exposed to 10% light intensity, the ultrastructure of chloroplasts was well developed, chloroplasts and starch grain size, the number of grana, and thylakoids all increased significantly, while the number of plastoglobules was significantly reduced. Relative distance phenotypic plasticity index analysis exhibited that P. arguta adapts to varying light environments predominantly by adjusting PPUE, Chl b, PNUE, chloroplast area, and the activity of PSII reaction centers. We proposed that P. arguta efficiently utilizes low light to reconfigure its energy metabolism by regulating its leaf structure, photosynthetic capacity, nutrient use efficiency, and chloroplast development.

Transcriptomic analysis of hub genes regulating albinism in light- and temperature-sensitive albino tea cultivars 'Zhonghuang 1' and 'Zhonghuang 2'.

Albino tea cultivars have high economic value because their young leaves contain enhanced free amino acids that improve the quality and properties of tea. Zhonghuang 1 (ZH1) and Zhonghuang 2 (ZH2) are two such cultivars widely planted in China; however, the environmental factors and molecular mechanisms regulating their yellow-leaf phenotype remain unclear. In this study, we demonstrated that both ZH1 and ZH2 are light- and temperature-sensitive. Under natural sunlight and low-temperature conditions, their young shoots were yellow with decreased chlorophyll and an abnormal chloroplast ultrastructure. Conversely, young shoots were green with increased chlorophyll and a normal chloroplast ultrastructure under shading and high-temperature conditions. RNA-seq analysis was performed for high light and low light conditions, and pairwise comparisons identified genes exhibiting different light responses between albino and green-leaf cultivars, including transcription factors, cytochrome P450 genes, and heat shock proteins. Weighted gene coexpression network analyses of RNA-seq data identified the modules related to chlorophyll differences between cultivars. Genes involved in chloroplast biogenesis and development, light signaling, and JA biosynthesis and signaling were typically downregulated in albino cultivars, accompanied by a decrease in JA-ILE content in ZH2 during the albino period. Furthermore, we identified the hub genes that may regulate the yellow-leaf phenotype of ZH1 and ZH2, including CsGDC1, CsALB4, CsGUN4, and a TPR gene (TEA010575.1), which were related to chloroplast biogenesis. This study provides new insights into the molecular mechanisms underlying leaf color formation in albino tea cultivars.

Effects of different light intensity on leaf color changes in a Chinese cabbage yellow cotyledon mutant.

Leaf color is one of the most important phenotypic features in horticultural crops and directly related to the contents of photosynthetic pigments. Most leaf color mutants are determined by the altered chlorophyll or carotenoid, which can be affected by light quality and intensity. Our previous study obtained a Chinese cabbage yellow cotyledon mutant that exhibited obvious yellow phenotypes in the cotyledons and the new leaves. However, the underlying mechanisms in the formation of yellow cotyledons and leaves remain unclear. In this study, the Chinese cabbage yellow cotyledon mutant 19YC-2 exhibited obvious difference in leaf color and abnormal chloroplast ultrastructure compared to the normal green cotyledon line 19GC-2. Remarkably, low-intensity light treatment caused turn-green leaves and a significant decrease in carotenoid content in 19YC-2. RNA-seq analysis revealed that the pathways of photosynthesis antenna proteins and carotenoid biosynthesis were significantly enriched during the process of leaf color changes, and many differentially expressed genes related to the two pathways were identified to respond to different light intensities. Remarkably, BrPDS and BrLCYE genes related to carotenoid biosynthesis showed significantly higher expression in 19YC-2 than that in 19GC-2, which was positively related to the higher carotenoid content in 19YC-2. In addition, several differentially expressed transcription factors were also identified and highly correlated to the changes in carotenoid content, suggesting that they may participate in the regulatory pathway of carotenoid biosynthesis. These findings provide insights into the molecular mechanisms of leaf color changes in yellow cotyledon mutant 19YC-2 of Chinese cabbage.

Antarctic benthic diatoms after 10 months of dark exposure: consequences for photosynthesis and cellular integrity.

Antarctic algae are exposed to prolonged periods of extreme darkness due to polar night, and coverage by ice and snow can extend such dark conditions to up to 10 months. A major group of microalgae in benthic habitats of Antarctica are diatoms, which are key primary producers in these regions. However, the effects of extremely prolonged dark exposure on their photosynthesis, cellular ultrastructure, and cell integrity remain unknown. Here we show that five strains of Antarctic benthic diatoms exhibit an active photosynthetic apparatus despite 10 months of dark-exposure. This was shown by a steady effective quantum yield of photosystem II (Y[II]) upon light exposure for up to 2.5 months, suggesting that Antarctic diatoms do not rely on metabolically inactive resting cells to survive prolonged darkness. While limnic strains performed better than their marine counterparts, Y(II) recovery to values commonly observed in diatoms occurred after 4-5 months of light exposure in all strains, suggesting long recovering times. Dark exposure for 10 months dramatically reduced the chloroplast ultrastructure, thylakoid stacking, and led to a higher proportion of cells with compromised membranes than in light-adapted cells. However, photosynthetic oxygen production was readily measurable after darkness and strong photoinhibition only occurred at high light levels (>800 µmol photons m-2 s-1). Our data suggest that Antarctic benthic diatoms are well adapted to long dark periods. However, prolonged darkness for several months followed by only few months of light and another dark period may prevent them to regain their full photosynthetic potential due to long recovery times, which might compromise long-term population survival.

Cytological, Physiological, and Transcriptomic Analyses of the Leaf Color Mutant Yellow Leaf 20 (yl20) in Eggplant (Solanum melongena L.)

Leaf color mutants are ideal materials for studying chlorophyll metabolism, chloroplast development, and photosynthesis in plants. We discovered a novel eggplant (Solanum melongena L.) mutant yl20 (yellow leaf 20) that exhibits yellow leaves. In this study, we compared the leaves of the mutant yl20 and wild type (WT) plants for cytological, physiological, and transcriptomic analyses. The results showed that the mutant yl20 exhibits abnormal chloroplast ultrastructure, reduced chlorophyll and carotenoid contents, and lower photosynthetic efficiency compared to the WT. Transcriptome data indicated 3267 and 478 differentially expressed genes (DEGs) between WT and yl20 lines in the cotyledon and euphylla stages, respectively, where most DEGs were downregulated in the yl20. Gene Ontology (GO) analysis revealed the “plastid-encoded plastid RNA polymerase complex” and the “chloroplast-related” terms were significantly enriched. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated that the significantly enriched DEGs were involved in flavone and flavonol biosynthesis, porphyrin and chlorophyll metabolism, etc. We speculated that these DEGs involved in significant terms were closely related to the leaf color development of the mutant yl20. Our results provide a possible explanation for the altered phenotype of leaf color mutants in eggplant and lay a theoretical foundation for plant breeding.

Maize requires arogenate dehydratase 2 for resistance to Ustilago maydis and plant development.

Maize (Zea mays) smut is a common biotrophic fungal disease caused by Ustilago maydis and leads to low maize yield. Maize resistance to U. maydis is a quantitative trait. However, the molecular mechanism underlying the resistance of maize to U. maydis is poorly understood. Here, we reported that a maize mutant caused by a single gene mutation exhibited defects in both fungal resistance and plant development. maize mutant highly susceptible to U. maydis (mmsu) with a dwarf phenotype forms tumors in the ear. A map-based cloning and allelism test demonstrated that 1 gene encoding a putative arogenate dehydratase/prephenate dehydratase (ADT/PDT) is responsible for the phenotypes of the mmsu and was designated as ZmADT2. Combined transcriptomic and metabolomic analyses revealed that mmsu had substantial differences in multiple metabolic pathways in response to U. maydis infection compared with the wild type. Disruption of ZmADT2 caused damage to the chloroplast ultrastructure and function, metabolic flux redirection, and reduced the amounts of salicylic acid (SA) and lignin, leading to susceptibility to U. maydis and dwarf phenotype. These results suggested that ZmADT2 is required for maintaining metabolic flux, as well as resistance to U. maydis and plant development in maize. Meanwhile, our findings provided insights into the maize response mechanism to U. maydis infection.

AmCBF1 activates the expression of GhClpR1 to mediate dark-green leaves in cotton (Gossypium hirsutum).

The transcription factor AmCBF1 deepens the leaf colour of transgenic cotton by binding to the promoter of the chloroplast development-related protein GhClpR1 to promote photosynthesis. The ATP-dependent caseinolytic protease (Clp protease) family plays a crucial role within chloroplasts, comprising several Clp proteins to maintain chloroplast homeostasis. At present, research on Clp proteins mainly focuses on Arabidopsis, leaving its function in other plants, particularly in crops, less explored. In this study, we overexpressed AmCBF1 from Ammopiptanthus mongolicus (A. mongolicus) in wild type (R15), and found a significant darkening of leaf colour in transgenic plants (L28 and L30). RNA-seq analysis showed an enrichment of pathways associated with photosynthesis. Subsequent screening of differentially expressed genes revealed a significant up-regulation of GhClpR1, a gene linked to chloroplast development, in the transgenic strain. In addition, GhClpR1 was consistently expressed in upland cotton, with the highest expression observed in leaves. Subcellular localization analysis revealed that the protein encoded by GhClpR1 was located in chloroplasts. Yeast one hybrid and dual luciferase experiments showed that the AmCBF1 transcription factor positively regulates the expression of GhClpR1. VIGs-mediated silencing of GhClpR1 led to a significant yellowing phenotype in the leaves. This was accompanied by a reduction in chlorophyll content, and microscopic examination of chloroplast ultrastructure revealed severe developmental impairment. Finally, yeast two-hybrid assays showed that GhClpR1 interacts with the Clp protease complex accessory protein GhClpT2. Our study provides a foundation for studying the function of the Clp protease complex and a new strategy for cultivating high-light-efficiency cotton resources.

Resting cell formation in the marine diatom Thalassiosira pseudonana.

Resting cells represent a survival strategy employed by diatoms to endure prolonged periods of unfavourable conditions. In the oceans, many diatoms sink at the end of their blooming season and therefore need to endure cold and dark conditions in the deeper layers of the water column. How they survive these conditions is largely unknown. We conducted an integrative analysis encompassing methods from histology, physiology, biochemistry, and genetics to reveal the biological mechanism of resting-cell formation in the model diatom Thalassiosira pseudonana. Resting-cell formation was triggered by a decrease in light and temperature with subsequent catabolism of storage compounds. Resting cells were characterised by an acidic and viscous cytoplasm and altered morphology of the chloroplast ultrastructure. The formation of resting cells in T. pseudonana is an energy demanding process required for a biophysical alteration of the cytosol and chloroplasts to endure the unfavourable conditions of the deeper ocean as photosynthetic organisms. However, most resting cells (> 90%) germinate upon return to favorable growth conditions.