Chloroplast Swelling Research Articles

Progressive heterogeneity of enlarged and irregularly shaped apicoplasts in P. falciparum persister blood stages after drug treatment.

Morphological modifications and shifts in organelle relationships are hallmarks of dormancy in eukaryotic cells. Communications between altered mitochondria and nuclei are associated with metabolic quiescence of cancer cells that can survive chemotherapy. In plants, changes in the pathways between nuclei, mitochondria, and chloroplasts are associated with cold stress and bud dormancy. Plasmodium falciparum parasites, the deadliest agent of malaria in humans, contain a chloroplast-like organelle (apicoplast) derived from an ancient photosynthetic symbiont. Antimalarial treatments can fail because a small fraction of the blood stage parasites enter dormancy and recrudesce after drug exposure. Altered mitochondrial-nuclear interactions in these persisters have been described for P. falciparum, but interactions of the apicoplast remained to be characterized. In the present study, we examined the apicoplasts of persisters obtained after exposure to dihydroartemisinin (a first-line antimalarial drug) followed by sorbitol treatment, or after exposure to sorbitol treatment alone. As previously observed, the mitochondrion of persisters was consistently enlarged and in close association with the nucleus. In contrast, the apicoplast varied from compact and oblate, like those of active ring stage parasites, to enlarged and irregularly shaped. Enlarged apicoplasts became more prevalent later in dormancy, but regular size apicoplasts subsequently predominated in actively replicating recrudescent parasites. All three organelles, nucleus, mitochondrion, and apicoplast, became closer during dormancy. Understanding their relationships in erythrocytic-stage persisters may lead to new strategies to prevent recrudescences and protect the future of malaria chemotherapy.

UV light and adaptive divergence of leaf physiology, anatomy, and ultrastructure drive heat stress tolerance in genetically distant grapevines.

The genetic basis of plant response to light and heat stresses had been unveiled, and different molecular mechanisms of leaf cell homeostasis to keep high physiological performances were recognized in grapevine varieties. However, the ability to develop heat stress tolerance strategies must be further elucidated since the morpho-anatomical and physiological traits involved may vary with genotype × environment combination, stress intensity, and duration. A 3-year experiment was conducted on potted plants of Sardinian red grapevine cultivars Cannonau (syn. Grenache) and Carignano (syn. Carignan), exposed to prolonged heat stress inside a UV-blocking greenhouse, either submitted to low daily UV-B doses of 4.63 kJ m-2 d-1 (+UV) or to 0 kJ m-2 d-1 (-UV), and compared to a control (C) exposed to solar radiation (4.05 kJ m-2 d-1 average UV-B dose). Irrigation was supplied to avoid water stress, and canopy light and thermal microclimate were monitored continuously. Heat stress exceeded one-third of the duration inside the greenhouse and 6% in C. In vivo spectroscopy, including leaf reflectance and fluorescence, allowed for characterizing different patterns of leaf traits and metabolites involved in oxidative stress protection. Cannonau showed lower stomatal conductance under C (200 mmol m-2 s-1) but more than twice the values inside the greenhouse (400 to 900 mmol m-2 s-1), where water use efficiency was reduced similarly in both varieties. Under severe heat stress and -UV, Cannonau showed a sharper decrease in primary photochemical activity and higher leaf pigment reflectance indexes and leaf mass area. UV-B increased the leaf pigments, especially in Carignano, and different leaf cell regulatory traits to prevent oxidative damage were observed in leaf cross-sections. Heat stress induced chloroplast swelling, plastoglobule diffusion, and the accumulation of secretion deposits in both varieties, aggravated in Cannonau -UV by cell vacuolation, membrane dilation, and diffused leaf blade spot swelling. Conversely, in Carignano UV-B, cell wall barriers and calcium oxalate crystals proliferated in mesophyll cells. These responses suggest an adaptive divergence among cultivars to prolonged heat stress and UV-B light. Further research on grapevine biodiversity, heat, and UV-B light interactions may give new insights on the extent of stress tolerance to improve viticulture adaptive strategies in climate change hotspots.

Stress-related physiological responses and ultrastructural changes in Hypoxis hemerocallidea leaves exposed to cadmium and aluminium

Hypoxis hemerocallidea is a medicinal plant containing hypoxoside (a pharmacologically active phytosterol diglucoside). This study evaluated the elemental composition in leaves of H. hemerocallidea treated with cadmium (Cd) and aluminium (Al) using scanning electron microscopy (SEM) combined with energy-dispersive X-ray spectroscopy (EDX). The impact of Cd and Al on photosynthetic pigments and performance, antioxidant activities and ultrastructure were also assessed. Corms of H. hemerocallidea were micropropagated, rooted and then exposed to varying concentrations of Cd, Al, and Cd + Al for six weeks. The SEM/EDX analysis indicated a two-fold increase in carbon content across all treated plants compared to the control. No/little Cd was detected in the leaves compared to a progressive increase in Al concentration with increasing Al treatment levels. This indicted that Al is more readily translocated to the shoots compared to Cd. Plants treated with Cd exhibited a significant decrease in total chlorophyll content accompanied by reduced photosynthetic performance and lower relative electron transport rates. Cd and Al exposure led to higher carotenoid, superoxide dismutase and malondialdehyde levels, indicating oxidative stress. Cd-treated plants displayed increased amylase activity and decreased carbohydrates content. Ultrastructural alterations occurred with exposure to Cd and Al, including abnormal swelling or disintegration of chloroplasts and thylakoid degeneration. An increase in starch grains and a decrease in plastoglobuli were also noted. In conclusion, this investigation provides evidence that both Cd and higher concentrations of Al exert detrimental effects on the ultrastructure, metabolism and photosynthetic performance of H. hemerocallidea, contributing to reduced growth and biological activity when stressed.

Polystyrene microplastic attenuated the toxic effects of florfenicol on rice (Oryza sativa L.) seedlings in hydroponics: From the perspective of oxidative response, phototoxicity and molecular metabolism

Antibiotics and microplastics (MPs) are two emerging pollutants in agroecosystems, however the effects of co-exposure to antibiotics and MPs remain unclear. The toxicity of florfenicol (FF) and polystyrene microplastics (PS-MPs) on rice seedlings was investigated. FF and PS-MPs caused colloidal agglomeration, which changed the environmental behavior of FF. FF inhibited rice growth and altered antioxidant enzyme (superoxide dismutase, peroxidase, and catalase) activities, leading to membrane lipid peroxidation; impaired photosynthetic systems, decreased photosynthetic pigments (Chlorophyll a, Chlorophyll b, and carotene), chlorophyll precursors (Proto IX, Mg-Proto IX, and Pchlide), photosynthetic and respiratory rates. The key photosynthesis related genes (PsaA, PsaB, PsbA, PsbB, PsbC, and PsbD) were significantly down-regulated. The ultrastructure of mesophyll cells was destroyed with chloroplast swelling, membrane surface blurring, irregular thylakoid lamellar structure, and number of peroxisomes increased. PS-MPs mitigated FF toxicity, and the IBR index values showed that 10 mg∙L−1 PS-MPs were more effective. Metabolomic analysis revealed that the abundance of metabolites and metabolic pathways were altered by FF, was greater than the combined "MPs-FF" contamination. The metabolism of amino acids, sugars, and organic acids were severely interfered. Among these, 15 metabolic pathways were significantly altered, with the most significant effects on phenylalanine metabolism and the citric acid cycle (p < 0.05).

Early infection response of fungal biotroph Ustilago maydis in maize.

Common smut, caused by Ustilago maydis (DC.) Corda, is a destructive fungal disease of maize worldwide; it forms large tumors, reducing corn yield and quality. However, the molecular defense mechanism to common smut in maize remains unclear. The present study aimed to use a leading maize inbred line Ye478 to analyze the response to U. maydis inoculation. The histological and cytological analyses demonstrated that U. maydis grew gradually to the host cells 6 h post-inoculation (hpi). The samples collected at 0, 3, 6, and 12 hpi were analyzed to assess the maize transcriptomic changes in response to U. maydis. The results revealed differences in hormone signaling, glycometabolism, and photosynthesis after U. maydis infection; specific changes were detected in jasmonic acid (JA), salicylic acid (SA), ethylene (ET), and abscisic acid (ABA) signaling pathways, glycolysis/gluconeogenesis, and photosystems I and II, probably related to defense response. MapMan analysis demonstrated that the differentially expressed genes between the treatment and control groups were clustered into light reaction and photorespiration pathways. In addition, U. maydis inoculation induced chloroplast swelling and damage, suggesting a significant effect on the chloroplast activity and subsequent metabolic process, especially hexose metabolism. A further genetic study using wild-type and galactinol-sucrose galactosyltransferase (gsg) and yellow-green leaf-1 (ygl-1) mutants identified that these two U. maydis-induced genes negatively regulated defense against common smut in maize. Our measurements showed the pathogen early-invasion process, and the key pathways of both chlorophyll biosynthesis and sugar transportation were critical modified in the infected maize line, thereby throwing a light on the molecular mechanisms in the maize-U. maydis interaction.

ROS as a key player in quinolone antibiotic stress on Arabidopsis thaliana: From the perspective of photosystem function, oxidative stress and phyllosphere microbiome

With the increasing use of antibiotics, their ecological impacts have received widespread attention. However, research on the toxicity of quinolone antibiotics is still limited, especially regarding the oxidative stress and phyllosphere of plants. In this study, the toxic effects of enrofloxacin, norfloxacin, and levofloxacin on Arabidopsis thaliana and their underlying mechanisms were investigated. The toxicity of the three quinolone antibiotics decreased in the following order: enrofloxacin > norfloxacin > levofloxacin. Physiological cellular changes, such as plasmolysis and chloroplast swelling, were observed using electron microscopy. Photosynthetic efficiency was inhibited with a decline in the effective photochemical quantum yield of photosystem II (Y(II)) and non-photochemical quenching (NPQ), indicating that quinolone antibiotics might reduce light energy conversion efficiency and excess light energy dissipation. Oxidative stress occurred in A. thaliana after quinolone antibiotic treatment, with an increase in reactive oxygen species (ROS) levels and malondialdehyde (MDA) content. High ROS levels stimulated the over-expression of superoxide-responsive genes for self-protection. Structural equation modeling (SEM) analysis showed that photosynthesis inhibition and cellular damage caused by oxidative stress were critical factors for growth inhibition, suggesting that the antioxidant response activated by ROS might be a potential mechanism. Furthermore, the diversity of the phyllospheric microbial communities decreased after enrofloxacin exposure. Additionally, specific microbes were preferentially recruited to the phyllosphere because of the higher ROS levels.

A comprehensive study of selenium and cerium oxide nanoparticles on mung bean: Individual and synergistic effect on photosynthesis pigments, antioxidants, and dry matter accumulation

In this study, the interaction effects of CeO2 NPs (250, 500 and 1000 mg L−1) and Se NPs (25, 50 and 75 mg L−1) were evaluated in mung bean (Vigna radiata). Single NPs and their combinations were foliar applied to 45-day old mung bean plants under greenhouse conditions. In each pot, a total volume of 100 mL of NPs suspension was sprayed on the plants shoot in two steps and one-week interval. After 94 days of growth, membrane degradation, antioxidant activity, photosynthetic pigments, and dry matter accumulation were assessed. At 250 and 500 mg CeO2-NPs L−1, there was partial increase of dry matter, stimulated activity of antioxidant enzymes (p ≤ 0.05), and reactive oxygen species (ROS). However, at 1000 mg L−1, CeO2-NPs caused strong accumulation of ROS (p ≤ 0.05), enlargement of starch granules and swelling of chloroplasts. In addition, at such concentration, there was accumulation of starch granules, reduction of photosynthetic pigments, biological nitrogen fixation, chlorosis, and a significant retardation in plant growth, compared with control, (p ≤ 0.05). Combination of Se-NPs (25 and 50 mg L−1) with 250 mg L−1 of CeO2 NPs decreased hydrogen peroxide, improved CAT, Chla, Chlb, and increased dry matter (p ≤ 0.05). At 1000 mg CeO2 NPs L−1, foliar spray of Se-NPs led to Ce accumulation in the cell wall and increased levels of SOD and proline (p ≤ 0.05). Results showed that 25 and 50 mg Se NPs L−1 ameliorate the stress of CeO2 NPs by upregulating photosynthesis pigments, antioxidants, and dry matter accumulation. Therefore, depending on the CeO2 NPs concentration, the mechanisms of Se NPs in modulating CeO2 NPs stress varied; low concentrations of Se NPs may strengthen the metabolism of legumes, and protect them against foliar toxicity of CeO2 NPs in semi-arid ecosystems.

Singlet Oxygen Leads to Structural Changes to Chloroplasts during their Degradation in the Arabidopsis thaliana plastid ferrochelatase two Mutant.

During stress, chloroplasts produce large amounts of reactive oxygen species (ROS). Chloroplasts also contain many nutrients, including 80% of a leaf's nitrogen supply. Therefore, to protect cells from photo-oxidative damage and to redistribute nutrients to sink tissues, chloroplasts are prime targets for degradation. Multiple chloroplast degradation pathways are induced by photo-oxidative stress or nutrient starvation, but the mechanisms by which damaged or senescing chloroplasts are identified, transported to the central vacuole and degraded are poorly defined. Here, we investigated the structures involved with degrading chloroplasts induced by the ROS singlet oxygen (1O2) in the Arabidopsis thaliana plastid ferrochelatase two (fc2) mutant. Under mild 1O2 stress, most fc2 chloroplasts appeared normal, but had reduced starch content. A subset of chloroplasts was degrading, and some protruded into the central vacuole via 'blebbing' structures. A 3D electron microscopy analysis demonstrated that up to 35% of degrading chloroplasts contained such structures. While the location of a chloroplast within a cell did not affect the likelihood of its degradation, chloroplasts in spongy mesophyll cells were degraded at a higher rate than those in palisade mesophyll cells. To determine if degrading chloroplasts have unique structural characteristics, allowing them to be distinguished from healthy chloroplasts, we analyzed fc2 seedlings grown under different levels of photo-oxidative stress. A correlation was observed among chloroplast swelling, 1O2 signaling and the state of degradation. Finally, plastoglobule (PG) enzymes involved in chloroplast disassembly were upregulated while PGs increased their association with the thylakoid grana, implicating an interaction between 1O2-induced chloroplast degradation and senescence pathways.

Manganese-induced oxidative stress, ultrastructural changes, and proteomics studies in rice plants.

Manganese (Mn) is an essential element for plant growth but it becomes phytotoxic at higher concentrations. The effect of Mn-excess in hydroponics medium was examined on growth, oxidative stress, and ultrastructural changes in chloroplasts and mitochondria as well proteomic alterations in rice (Oryza sativa L.) seedlings. Seedlings grown with 1mM and 2mM Mn in nutrient medium for 8days showed decline in length and fresh biomass, and decline in net photosynthetic rate, transpiration rate, and stomatal conductance. Shoots of the seedlings had higher Mn content than roots. Mn-treated seedlings showed increased production of O2·-, H2O2, and .OH, increased lipid peroxidation, increased carbonylation of proteins, and increased proteolytic activity compared to untreated seedlings. Mn-treated seedlings showed disorganization and swelling of chloroplasts with appearance of plastoglobuli in TEM images and deformity in shape of mitochondria. Using confocal microscopy depolarization of mitochondrial membrane was observed marked by green fluorescence of JC-1 dye monomers in Mn-treated roots. Proteomics studies from leaves of Mn-treated seedlings involving 2DE and PDQuest analysis showed differential expression of 23 proteins, among which MALDI-TOF/TOF mass spectrometry analysis revealed Mn-led downregulation of photosynthesis-related proteins, namely oxygen-evolving complex protein associated with PSII, PAP-3, enzyme involved in protein folding peptidyl-prolyl cis-trans isomerase (PPIase) and carbohydrate metabolizing enzymes hydrolase, fructose-bisphosphate aldolase, transketolase, and isocitrate dehydrogenase, whereas ATP-dependent Clp protease, peroxidase, and nucleic acid-binding proteins were downregulated due to Mn treatment. Results indicate that Mn-excess inhibits growth of rice plants with induction of oxidative stress, causing structural alterations in chloroplasts, mitochondria, inhibiting photosynthesis, and downregulating many photosynthesis and carbohydrate metabolism-related proteins.

Transcriptome analysis of mechanisms and candidate genes associated with cucumber response to cucumber alternaria leaf spot infection

Cucumber alternaria leaf spot is a fungal disease of cucumber (Cucumis sativus L.) caused by the pathogen Alternaria cucumerina (Ell et Ev.) Elliott. RNA-seq was used to reveal plants response to Alternaria cucumerina infection at 2 and 6 days post-inoculation in two cucumber varieties—a resistant variety, D1322; and a susceptible one, BJ204. A total of 2015 and 162 differentially expressed genes were identified in D1322, whereas 5276 and 307 were identified in Beijing204, at 2 and 6 days post-inoculation, respectively. Gene Ontology and KEGG pathway enrichment analysis of differentially exressed genes indicated that the significantly enriched GO terms and pathways were similar between D1322 and BJ204. Differentially expressed genes associated with the significantly enriched pathways: phenylpropanoid biosynthetic, phenylalanine metabolism and glutathione metabolism pathways, were mostly up-regulated. Differentially expressed genes related to the photosynthesis performance were down-regulated just in BJ04. Chloroplast swelling and grana lamellae disintegration were also observed in BJ204 with transmission electron microscopy. Many up-regulated transcription factors were induced following Alternaria cucumerina infection. This study provides valuable information on the molecular basis of cucumber–Alternaria cucumerina interaction mechanisms and offers candidate genes related to cucumber response to Alternaria cucumerina infection.

Effects of ketoprofen on rice seedlings: Insights from photosynthesis, antioxidative stress, gene expression patterns, and integrated biomarker response analysis

Pharmacologically active compounds found in reclaimed wastewater irrigation or animal manure fertilizers pose potential risks for agriculture. The mechanism underlying the effects of ketoprofen on rice (Oryza sativa L.) seedlings was investigated. The results showed that low concentrations (0.5 mg L−1) of ketoprofen slightly stimulate growth of rice seedlings, while high concentrations can significantly inhibit growth by reducing biomass and causing damage to roots. Ketoprofen affects photosynthetic pigment content (Chla, Chlb, and carotenoids) and chlorophyll synthesis gene (HEMA, HEMG, CHLD, CHLG, CHLM, and CAO) expression. Fluorescence parameters such as minimum fluorescence (F0), maximum fluorescence (Fm), variable fluorescence (Fv), potential photosynthetic capacity (Fv/F0), maximum quantum efficiency of PSII photochemistry (Fv/Fm), electron transfer rate (ETR), and Y(II), Y(NPQ), Y(NO) values were affected, showing photosynthetic electron transfer was blocked. Active oxygen radical (O2•−and H2O2), malondialdehyde and proline content increased. Superoxide dismutase, catalase and ascorbate peroxidase activities, glutathione content and antioxidant-related gene (FSD1, MSD1, CSD1, CSD2, CAT1, CAT2, CAT3, APX1, APX2) expression were induced. Higher integrated biomarker response values of eight oxidative stress response indexes were obtained at higher ketoprofen concentrations. Ultrastructure observation showed that ketoprofen causes cell structure damage, chloroplast swelling, increase in starch granules, and reduction in organelles. This study provides some suggested toxicological mechanisms and biological response indicators in rice due to stress from pharmacologically active compounds.

Comparative Cytological and Gene Expression Analysis Reveals Potential Metabolic Pathways and Target Genes Responsive to Salt Stress in Kenaf (Hibiscus cannabinus L.)

Salt stress represents the most unfortunate abiotic stress factor in agricultural areas worldwide and contributes a severe reduction in growth, development, and yield of crop plants. Kenaf is a salt-tolerant crop. However, molecular mechanisms of salt stress tolerance in kenaf are not well elucidated. In the present study, seed germination parameters and cytological and transcriptome responses of leaves from two cultivars of kenaf (salt-tolerant P3A and salt-sensitive P3B) were analyzed between 250 and 0 mM NaCl treatments. Results indicated that under salt stress cultivar P3A had higher seed germination energy, germination index, and germination percentage with the less relative rate of salt damage than cultivar P3B. Furthermore, cytological studies indicated swelling and vacuolization of chloroplast and thylakoid in cultivar P3A under the influence of salt stress; however, chloroplast and thylakoid were degraded in cultivar P3B. Transcriptome analysis generated 8466 and 9333 differentially expressed unigenes (DEGs) (FDR ≤ 0.05 and log2FC ≥ 1) between salt-stressed and control treatments in cultivar P3A and cultivar P3B, respectively. Eight DEGs were selected randomly for quantitative real-time PCR to test the reliability of transcriptome sequencing results. Several genes encoding transcription factors WRKY, AP2/EREBP, and Hsfs families and genes like LEA, PsbA, PK, GSTs, NPR1, and TGA were induced under salt stress, and expression levels were higher in the cultivar P3A compared to cultivar P3B. The GO enrichment and KEGG pathway analysis suggested that these DEGs were related to ionic homeostasis and transport, osmotic adjustment, water deficit response, antioxidants, ROS scavenging, cellular membranes protection, photo-damage repairing cycle of photosystem II, thylakoid part, plant hormone signal transduction pathway, and may be related to salt tolerance in kenaf.

Comparative molecular and metabolic responses of wheat seedlings (Triticum aestivum L.) to the imazethapyr enantiomers S-IM and R-IM

The enantioselective effects of imazethapyr (IM) enantiomers on wheat seedlings in a hydroponic medium were studied. R-IM at 0.05mg/L exerted a stronger inhibitory effect on shoot weight and root weight than 0.05mg/L S-IM, suggesting that R-IM more severely inhibited growth. Oxidative damage, based on the anthocyanin content, malondialdehyde (MDA) content, antioxidant enzyme activities and transcript levels of antioxidant enzyme genes, were studied together with the cellular ultrastructure of wheat leaves. The anthocyanin and MDA contents in the R-IM treatment group were significantly increased compared with those in the control group, but no significant changes were observed in the S-IM treatment group. The antioxidant enzyme activities of CAT and SOD were inhibited by 0.32- and 0.73-fold, respectively, in the 14day R-IM treatment group compared to those in the control. However, the transcript levels of antioxidant enzyme genes, including CuZnSOD, POD and CAT, were downregulated in the 14day R-IM exposure group, but those of DHAR were not. The number and size of starch granules increased and chloroplast swelling was observed in wheat leaf cells after R-IM exposure, which showed that photosynthetic functions were potentially disturbed. These results directly or indirectly imply that R-IM exposure causes more oxidative stress and exerts a stronger negative effect on wheat than S-IM. A metabolomics approach revealed that the tricarboxylic acid cycle was heavily suppressed by R-IM treatment. Some amino acids (proline, threonine, lysine, valine) were increased by only the R-IM treatment, indicating the activation of antioxidant pathways. The decrease in a series of fatty acids implied that the cell membrane composition changed in response to R-IM. These results provide a deeper understanding of the enantioselective effects of IM enantiomers on the molecular and metabolic responses in wheat seedlings.

Effect of differently methyl-substituted ionic liquids on Scenedesmus obliquus growth, photosynthesis, respiration, and ultrastructure

Concerns have been raised regarding the ecotoxicity of ionic liquids (ILs) owing to their wide usage in numerous fields. Three imidazolium chloride ILs with different numbers of methyl substituents, 1-decyl-imidazolium chloride ([C10IM]Cl), 1-decyl-3-methylimidazolium chloride ([C10MIM]Cl), and 1-decyl-2,3-dimethylimidazolium chloride ([C10DMIM]Cl), were examined to assess their effects on growth, photosynthesis pigments content, chlorophyll fluorescence, photosynthetic and respiration rate, and cellular ultrastructure of Scenedesmus obliquus. The results showed that algal growth was significantly inhibited by ILs treatments. The observed IC50,48h doses were 0.10 mg/L [C10IM]Cl, 0.01 mg/L [C10MIM]Cl, and 0.02 mg/L [C10DMIM]Cl. The chlorophyll a, chlorophyll b, and total chlorophyll content declined, and the chlorophyll fluorescence parameters, minimal fluorescence yield (F0), maximal fluorescence yield (Fm), maximum quantum yield of PSII photochemistry (Fv/Fm), effective quantum yield of PSII [Y(II)], non-photochemical quenching (NPQ) and non-photosynthetic losses yield [Y(NO)] were notably affected by ILs in a dose-dependent manner. ILs affected the primary photosynthetic reaction, impaired heat dissipation capability, and diminished photosynthetic efficiency, indicating negative effects on photosystem II. The photosynthetic and respiration rates of algal cells were also reduced due to the ILs treatments. The adverse effects of ILs on plasmolysis and chloroplast deformation were examined using ultrastructural analyses; chloroplast swelling and lamellar structure almost disappeared after the [C10MIM]Cl treatment, and an increased number of starch grains and vacuoles was observed after all ILs treatments. The results indicated that one-methyl-substituted ILs were more toxic than non-methyl-substituted ILs, which were also more toxic than di-methyl-substituted ILs. The toxicity of the examined ILs showed the following order: [C10IM]Cl < [C10DMIM]Cl ≤ [C10MIM]Cl.

Tolerance mechanism of Triarrhena sacchariflora (Maxim.) Nakai. seedlings to lead and cadmium: Translocation, subcellular distribution, chemical forms and variations in leaf ultrastructure

Hydroponic experiments were conducted to assess the accumulation, translocation, and chemical forms of lead (Pb) and cadmium (Cd) in the roots, stems, and leaves of Triarrhena sacchariflora seedlings and the associated variation in leaf ultrastructure. The leaves and leaf ultrastructure showed no significant symptoms of toxicity with 0.05 mM Pb or 0.01 mM Cd exposure for 10d. Chlorosis and wilting were observed in leaves when the Pb and Cd concentration was higher than 0.1 and 0.05 mM in the medium, respectively, as demonstrated by severe ultrastructural modifications at higher concentration in the leaves, such as plasmolysis, cell wall detachment, chloroplast swelling, nuclear condensation, and even nuclear fragmentation. The Pb and Cd concentrations in the roots was significantly higher than those in the stems and leaves. This indicated low Pb and Cd translocation from the roots to the aboveground parts. Subcellular distribution analysis showed that the majority of Pb and Cd was bound to the cell wall, especially in the roots, indicating that the cell wall likely constitutes a crucial storage site for Pb and Cd. This mechanism decreases the translocation of Pb and Cd across membranes and is more effective than vacuolar compartmentation. The majority of Pb and Cd exited in form of insoluble Pb/Cd-pectate or -oxalate complexes in the plant. In conclusion, higher concentrations of Pb or Cd induced premature senescence. High Pb and Cd enrichment was observed in the roots, which decreased the translocation of Pb and Cd from the roots to the aboveground tissues. The immobilization of Pb or Cd by the cell wall is important for plant detoxification and can protect protoplasts from Pb or Cd toxicity. Pb and Cd mainly existed in insoluble Pb/Cd-phosphate or -oxalate complexes, exhibiting low activity and thereby limiting symplastic transport and suppressing toxicity.

Effect of imidazolium-based ionic liquids with varying carbon chain lengths on Arabidopsis thaliana: Response of growth and photosynthetic fluorescence parameters

As green and novel solvents, ionic liquids (ILs) are popular in many industries, which may threaten ecosystems. The effects of three imidazolium-based ILs with different alkyl chain lengths, including 1-octyl-3-methylimidazolium chloride ([C8mim]Cl), 1-decyl-3-methylimidazolium chloride ([C10mim]Cl), and 1-dodecyl-3-methylimidazolium chloride ([C12mim]Cl) on growth and photosystem of Arabidopsis thaliana were investigated. Root length, fresh weight, cell membrane permeability, and chlorophyll content of whole plant were significantly affected by ILs. Vein clearing, leaf chlorosis, and browning on the A. thaliana leaf abaxial surface occurred, with a dose-response relationship. The effect of ILs on whole plant increased with alkyl chain lengths. Chlorophyll fluorescence parameters of photosynthetic system II (PSII) were all affected in [C8mim]Cl and [C10mim]Cl treatments, electron-transfer was blocked, and photochemical energy conversion was damaged. There were no significant changes in chlorophyll fluorescence of newly-growing leaves in [C12mim]Cl treatment, but has severe effect on aged leaves. The number and size of starch granules and osmiophilic globules increased, plasmolysis and the chloroplast swelling occurred in [C8mim]Cl, [C10mim]Cl treatments and on aged leaves in [C12mim]Cl treatment, but no significant damages occurred on newly-growing leaves of [C12mim]Cl treatment, perhaps due to plant self-protection of plant. The results indicating the appropriate use of ILs is needed.

Selective Elimination of Membrane-Damaged Chloroplasts via Microautophagy.

Plant chloroplasts constantly accumulate damage caused by visible wavelengths of light during photosynthesis. Our previous study revealed that entire photodamaged chloroplasts are subjected to vacuolar digestion through an autophagy process termed chlorophagy; however, how this process is induced and executed remained poorly understood. In this study, we monitored intracellular induction of chlorophagy in Arabidopsis (Arabidopsis thaliana) leaves and found that mesophyll cells damaged by high visible light displayed abnormal chloroplasts with a swollen shape and 2.5 times the volume of normal chloroplasts. In wild-type plants, the activation of chlorophagy decreased the number of swollen chloroplasts. In the autophagy-deficient autophagy mutants, the swollen chloroplasts persisted, and dysfunctional chloroplasts that had lost chlorophyll fluorescence accumulated in the cytoplasm. Chloroplast swelling and subsequent induction of chlorophagy were suppressed by the application of exogenous mannitol to increase the osmotic pressure outside chloroplasts or by overexpression of VESICLE INDUCING PROTEIN IN PLASTID1, which maintains chloroplast envelope integrity. Microscopic observations of autophagy-related membranes showed that swollen chloroplasts were partly surrounded by autophagosomal structures and were engulfed directly by the tonoplast, as in microautophagy. Our results indicate that an elevation in osmotic potential inside the chloroplast due to high visible light-derived envelope damage results in chloroplast swelling and serves as an induction factor for chlorophagy, and this process mobilizes entire chloroplasts via tonoplast-mediated sequestering to avoid the cytosolic accumulation of dysfunctional chloroplasts.

English

Water and light are the most important environements for plants’ growth. These environemts are critical factors needed not only for the survival of plants but also their production. When plants are exposed to drought condition, they change in their anatomical, physiological and biochemical properties. Drought affects plants wildly from their cell structure to growth. It causes higher plastoglobuli, lower starch grain, distortion of thylakoids, disrupted grana and swelling of chloroplast. Plants grown under enhanced light, have increased palisade parenchyma, thicker leaf, higher biomass, increased photosynthesis, lower contents of chlorophyll, carotenoid and nitrogen. Keywords: Drought, plant, shade, water

Synergism of herbicide toxicity by 5-aminolevulinic acid is related to physiological and ultra-structural disorders in crickweed (Malachium aquaticum L.)

Selection of effective herbicides to control weeds has been one of the major objectives of scientists. This study determines the differential tolerance or susceptibility of crickweed (Malachium aquaticum L.) to various concentration combinations of 5-aminolevulinic acid (ALA) (1, 10 and 100mg/L) and propyl 4-(2-(4,6-dimethoxypyrimidin-2-yloxy)benzylamino)benzoate (ZJ0273) (100, 200, and 500mg/L). ALA was applied as pre- and post-treatment alone or in combination with ZJ0273. Results showed that ZJ0273 stress alone imposed negative effects on M. aquaticum seedling's growth, net photosynthetic rates and SPAD values, and the rate of decline was consistently increased with the increase in ZJ0273 concentration. The ZJ0273 treatment showed a gradual decrease in the activities of antioxidant enzymes peroxidase (POD), superoxide dismutase (SOD), and ascorbate peroxidase (APX), and increase in the accumulation of malondialdehyde (MDA). Changes in chloroplast swelling, increased number of plastoglobuli, disruption of thylakoid, disintegrated mitochondria and turbid nucleoplasm were noticed. Moreover, SDS-PAGE analysis of total proteins revealed that herbicide stress in the leaves was associated with the decrease or disappearance of some protein bands. Further, two-dimensional gel electrophoresis (2-DE) results showed that proteins in different spots were classified into three types for M. aquaticum. These results indicate that the combined treatment of ALA and ZJ0273 synergizes the herbicide toxicity which is different from its independent effects on M. aquaticum and thus, could improve weed control efficacy.

Effect of exogenous 24-epibrassinolide on chlorophyll fluorescence, leaf surface morphology and cellular ultrastructure of grape seedlings (Vitis vinifera L.) under water stress

The effects of 24-epibrassinolide (EBR) on chlorophyll fluorescence, leaf surface morphology and cellular ultrastructure of grape seedlings (Vitis vinifera L.) under water stress were investigated. The grape seedlings were subjected to 10 % (w/v) polyethylene glycol (PEG-6000) and treated with 0.05, 0.10 or 0.20 mg L−1 EBR, respectively. EBR application increased chlorophyll contents, the effective photochemical quantum yield of PSII, maximum photochemical efficiency of PSII, maximal fluorescence and non-photochemical quenching coefficient under water stress in each concentration. Compared with water stress control, higher stomatal density and stomatal length were observed in young leaves under EBR treatments, but not in mature leaves. In-depth analysis of the ultrastructure of leaves indicated that water stress induced disappearance of nucleus, chloroplast swelling, fractured mitochondrial cristae and disorder of thylakoid arrangement both in young leaves and mature leaves. However, EBR application counteracted the detrimental effects of water stress on the structure of the photosynthetic apparatus better in young leaves than in mature leaves. Compared to the other treatments, treatment of 0.10 mg L−1 EBR had best ameliorative effect against water stress. These results suggested that exogenous EBR could alleviate water stress-induced inhibition of photosynthesis on grape possibly through increasing chlorophyll content, lessening the stomatal and non-stomatal limitation of photosynthesis performance.