Disintegration Of Chloroplasts Research Articles

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.

Involvement of BcERF109 in 1-MCP-delayed yellowing of pak choi (Brassica rapa subsp. Chinensis) via mitigation of major chlorophyll-degrading gene expression during storage

Leaf yellowing is a typical senescence symptom in postharvest pak choi (Brassica rapa subsp. Chinensis). Ethylene response factors (ERFs) are considered important regulators of leaf senescence. This study explored the likely regulatory effects of BcERF on pak choi yellowing via 1-methylcyclopropene (1-MCP) treatments. We identified an ERF transcription factor (TF), BcERF109, which may be involved in ethylene-induced pak choi yellowing. Treatment with 5.0 µL L–1 1-MCP suppressed the transcripts of chlorophyll catabolism genes (BcPPH1, BcRCCR, BcSGR2, and BcNYC1), inhibited decreases in chlorophyll content and chlorophyll fluorescence parameters, delayed chloroplast disintegration, and eventually mitigated yellowing. The treatment also blocked ethylene production and impeded the activities and transcripts of ethylene synthesis enzymes (ACC oxidase [ACO] and 1-aminocyclopropene-1-carboxylate [ACC] synthase [ACS]). In contrast, treatment with 10.0 µL L–1 ethylene accelerated pak choi yellowing, as indicated by greater ethylene production, activities and transcripts of ACO and ACS, and expression of chlorophyll catabolism genes, as well as reductions in chlorophyll content and fluorescence parameters. BcERF109 was identified as a nuclear-localised protein; its transcription levels were downregulated by 1-MCP and upregulated by ethylene. Molecular biology assays revealed that BcERF109 enhanced the transcriptional capacities of BcPPH1, BcRCCR, BcNYC1, and BcSGR2 by binding to their promoters. The transient overexpressing of BcERF109 in tobacco leaves stimulated the expression of endogenous genes (BcPPH1, BcRCCR, BcNYC1, BcSGR2, and BcERF109), leading to accelerated chlorophyll degradation and leaf yellowing. The application of 1-MCP mitigated pak choi yellowing by influencing ethylene synthesis and modulating the regulatory effects of BcERF019 on genes associated with chlorophyll catabolism. These results revealed a new regulatory mechanism involving ERF TFs in 1-MCP-induced delayed yellowing of green vegetables.

Chronic toxicity of pharmaceuticals to the benthic green alga Closterium ehrenbergii

Pharmaceuticals in the environment have emerged to a topic of global concern. Since these substances are designed to be biologically active, hazardous effects on non-target organisms are frequently reported. Here, the effects of five pharmaceuticals, one radiocontrast agent, and one degradation product on the freshwater green alga Closterium ehrenbergii were evaluated after chronic exposure of 168 h. Growth and maximum quantum yield (FV/FM) were used as endpoints and complemented by the assessment of morphology and chlorophyll fluorescence. We found that the tested antibiotics Ciprofloxacin and Ofloxacin impaired chloroplast integrity, resulting in a reduction of FV/FM from 0.1 mg/L. The disintegration of chloroplasts at higher concentrations (c = 0.3 and 0.8 mg/L, respectively) was visualized by brightfield and fluorescence microscopy. In contrast, Sulfamethoxazole interfered with cell division, leading to malformation of cells from 0.8 mg/L. Furthermore, the antibiotics exhibited a latency period of 72 h after which they started to reveal their true effects. Therefore, the importance of long-term toxicity testing is outlined in order to avoid underestimation of toxic effects of pharmaceuticals. Based on the EC10 values obtained, the antibiotics were considered to meet the criteria for classification as toxic to aquatic life with long lasting effects. The other test substances were found to exert no effects on C. ehrenbergii or only at very high concentrations and were classified as nontoxic.

Scrubber Filter in the Phosphate Fertilizer Factory Reduces Fluorine Emission and Accumulation in Corn

Fluorine (F) produced from the fertilizer factory occurs in the process of phosphate fertilizer production, using sulfur and phosphate rocks as raw materials. Technologies to control atmospheric pollution with F should be adopted to reduce the impact on agricultural production. This study has the hypothesis that the emission of F, derived from the chimneys of fertilizer factories, is influencing the quality of corn (Zea mays L.) and increasing the F levels in the soil and plants. The objective of the study was to monitor the contents of F in corn leaves and soil in properties located close to the fertilizer production industry (between 1.5 and 2.0 km) before and after the installation of scrubber filters in the chimneys of the factory. A field study was carried out during the 2020/2021 harvest to evaluate the contents of F in corn plants and soil. Results showed that the scrubber filter installation represented a F reduction average of 92% in leaves comparing the average before the scrubber filter installation. Corn showed symptoms of F toxicity, such as leaf chlorosis, caused by the disintegration of chloroplasts, inhibition of photosynthesis, and others. In addition, there was a reduction of 40% (from the first to the second collecting) and 75% (from the first to the third collecting) in the levels of F in the soil after the scrubber filter installation. Based on the results, we conclude that the implementation of a scrubber filter is an optimal alternative to reduce F levels in corn leaves and the soil in properties located close to a fertilizer factory.

Melatonin delays leaf senescence and improves cucumber yield by modulating chlorophyll degradation and photoinhibition of PSII and PSI

Decreases in chlorophyll and photosynthetic efficiency co-occur during aging and leaf senescence. Here, we report the positive functions of melatonin (MT) in delaying leaf senescence in cucumber plants. The results showed that natural leaf senescence induced the accumulation of endogenous MT and that spraying MT delayed leaf senescence in a concentration-dependent manner. Moreover, application of MT and transient overexpression of CsASMT, the key gene in MT synthesis, both suppressed leaf senescence simulated by darkness. This was evidenced by a decrease in expression of the senescence-related gene SAG12 and the cell death-related gene PDCD, an increase in chlorophyll content in terms of downregulation of chlorophyll-degradation genes and normal chloroplast structure, and higher activity of PSI and PSII reaction centers, electron transfer efficiency and photosynthesis-associated protein levels, compared with these measures in the H2O-treated seedlings. In addition, the data from field spraying experiments demonstrated that MT indeed alleviated the decline in chlorophyll content and increased photosynthetic efficiency in both functional and senescent leaves, which ultimately promoted the growth and yield of cucumber in the solar greenhouse. Together, these results indicate that MT acts as a positive regulator to downregulate SAG12 and PDCD mRNA abundances and alleviate damage to the photosynthetic apparatus by suppressing chloroplast disintegration and photosynthesis-associated protein degradation during leaf senescence.

Near-freezing temperature storage improves shelf-life and suppresses chilling injury in postharvest apricot fruit (Prunus armeniaca L.) by regulating cell wall metabolism

To avoid chilling injury (CI) of apricots during storage, 1–2 °C and 4–6 °C storage as controls, the relationship between changes in cell wall characteristics and the occurrences of chilling injury under near-freezing temperature (NFT) storage was studied. NFT-stored improved apricots quality and inhibited CI index, membrane permeability and malondialdehyde (MDA) content; This also significantly inhibited the activity of cell wall modifying enzymes, delaying the solubilization of water-soluble pectin (WSP) and the degradation of cellulose. Transmission electron microscopy (TEM) imaging revealed that the density of the middle lamella in stored for 49 d was higher NFT than controlled temperature, delaying cell wall and chloroplast disintegration. Additionally, NFT-stored has no CI during the shelf life, and can be normal after ripening, maintaining higher commodity rate and sensory characteristics. These conclusions show that NFT storage can effectively improve the cold resistance of apricot fruit.

Toxicity assessments of acrylamide in aquatic environment using two algae Nitzschia closterium and Scenedesmus quadricauda.

The expanding production and widespread application of acrylamide caused inevitable release to aquatic ecosystems. Contrary to its extensive attention to human and animal health, the hazards of acrylamide to the aquatic primary productivity have been rarely studied. The potential effects of acrylamide on the marine algae (Nitzschia closterium) and the limnetic algae (Scenedesmus quadricauda) were investigated by monitoring cell abundance, total chlorophyll content, maximum photosystem II (PSII) quantum yield (Fv/Fm), and reactive oxygen species (ROS). The growth of two algae was significantly inhibited by acrylamide. The 96h EC50 of acrylamide on N. closterium and S. quadricauda were 5.50mgL-1 and 45.3mgL-1, and no observed effect concentration (NOEC) were 1.07mgL-1 and 6.97mgL-1, respectively. After 96h exposure to 50mgL-1 of acrylamide, the total chlorophyll content declined to approximate 18% (N. closterium) and 48% (S. quadricauda), and Fv/Fm was observed to be 0.35 and 0.32 for N. closterium and S. quadricauda, respectively. ROS was significantly increased following higher exposure concentrations, and its levels increased around 2.1-fold and 1.4-fold following exposure to 5mgL-1 of acrylamide. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) showed the visible cell plasmolysis, rupture of the plasma membrane, cell vacuolization, and disintegration of chloroplasts of the algae caused acrylamide.

Comparative Proteomic Analysis by iTRAQ Reveals that Plastid Pigment Metabolism Contributes to Leaf Color Changes in Tobacco (Nicotiana tabacum) during Curing.

Tobacco (Nicotiana tabacum), is a world’s major non-food agricultural crop widely cultivated for its economic value. Among several color change associated biological processes, plastid pigment metabolism is of trivial importance in postharvest plant organs during curing and storage. However, the molecular mechanisms involved in carotenoid and chlorophyll metabolism, as well as color change in tobacco leaves during curing, need further elaboration. Here, proteomic analysis at different curing stages (0 h, 48 h, 72 h) was performed in tobacco cv. Bi’na1 with an aim to investigate the molecular mechanisms of pigment metabolism in tobacco leaves as revealed by the iTRAQ proteomic approach. Our results displayed significant differences in leaf color parameters and ultrastructural fingerprints that indicate an acceleration of chloroplast disintegration and promotion of pigment degradation in tobacco leaves due to curing. In total, 5931 proteins were identified, of which 923 (450 up-regulated, 452 down-regulated, and 21 common) differentially expressed proteins (DEPs) were obtained from tobacco leaves. To elucidate the molecular mechanisms of pigment metabolism and color change, 19 DEPs involved in carotenoid metabolism and 12 DEPs related to chlorophyll metabolism were screened. The results exhibited the complex regulation of DEPs in carotenoid metabolism, a negative regulation in chlorophyll biosynthesis, and a positive regulation in chlorophyll breakdown, which delayed the degradation of xanthophylls and accelerated the breakdown of chlorophylls, promoting the formation of yellow color during curing. Particularly, the up-regulation of the chlorophyllase-1-like isoform X2 was the key protein regulatory mechanism responsible for chlorophyll metabolism and color change. The expression pattern of 8 genes was consistent with the iTRAQ data. These results not only provide new insights into pigment metabolism and color change underlying the postharvest physiological regulatory networks in plants, but also a broader perspective, which prompts us to pay attention to further screen key proteins in tobacco leaves during curing.

ES5 is involved in the regulation of phosphatidylserine synthesis and impacts on early senescence in rice (Oryza sativa L.)

Leaf senescence, which affects plant growth and yield in rice, is an ideal target for crop improvement and remarkable advances have been made to identify the mechanism underlying this process. We have characterized an early senile mutant es5 (early leaf senescence 5) in rice exhibiting leaf yellowing phenotype after the 4-leaf stage. This phenotype was confirmed by the higher accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), the disintegration of chloroplasts, reduction in chlorophyll content and photosynthetic rate and up-regulation of senescence-associated genes (SAGs) like Osh36, OsI57, and OsI85. Positional cloning revealed that the es5 phenotype is the result of one base substitution in ES5, encoding phosphatidylserine synthase (PSS) family protein, which is involved in the base-exchange type reaction to synthesize the minor membrane phospholipid phosphatidylserine. Functional complementation of ES5 in the es5 plants completely restored the wild-type phenotype. Ultra-high-performance liquid chromatography (UHPLC) analysis showed that es5 plants had increased levels of phosphatidylserine (PS) and decreased level of phosphatidylcholine (PC). These results provide evidence about the role of PS in rice leaf senescence.

Boron toxicity induced specific changes of cell ultrastructure and architecture of components in leaf center and tip of trifoliate orange [Poncirus trifoliata (L.) Raf.

The distribution of boron (B) in leaves is far from uniform, and tolerance to B toxicity should be varied in different portions of an entire leaf. Here, according to the order and degree of leaf chlorosis, a whole leaf blade of trifoliate orange [Poncirus trifoliata (L.) Raf.] rootstock was divided into two segments-leaf tip and leaf center, and transmission electron microscope (TEM) and fourier transform infrared spectroscopy (FTIR) were used to obtain more detailed information on the cell ultrastructure and component architecture of the two leaf segments under B toxicity. Results revealed that B toxicity led to alterations in pectin network crosslinking structure of leaf tip and destruction of cell wall integrity. Moreover, B toxicity altered protein structure and decreased protein content, while increased carbohydrate content in the two leaf segments, especially in leaf tip. Excess B supply reduced the cellulose content in leaf tip but increased in leaf center. TEM micrographs exhibited chloroplast disintegration and plastoglobulus accumulation in cells of two different leaf sections of B-toxicity plants, with less pronounced changes in leaf center. Furthermore, B toxicity only induced accumulation of starch grains in cells of leaf center. Overall results indicated that the B-toxic-induced biochemical changes of the cell ultrastructure and component architecture greatly differed in leaf tip and center. This study facilitates a better understanding of structural changes in different leaf portions of P. trifoliata under B toxicity stress and provides new ideas for further research on other elements in different plant leaf portions.

The resistance of the covering tissues of apple shoots depending on thier hydration and the ambient temperature

The article considers the questions of stability of epithelialtissues of Apple shoots depending on their hydrationand the ambient temperature. Studies have shown thatwith increased exposure of annual shoots of Apple trees inintensive sunlight disintegration of chloroplasts increased.It starts in the cells of collenchyma and extended towardsthe parenchyma. The location of Apple shoots in the most litarea led to an increase of actinobacilli of chlorophyll. Thedifference in the destroyed chloroplasts in the bark of shootsof the peripheral part (on the South side of the tree) andthe center of the crown amounted to 34%. Damage chlorophyllosecells to intense light was due to excess light energy,newsrevue be disposed in photosynthesis as the resultof photodynamic effect. In embodiments, where the shootswere exposed to strong light in a cold room, bark damageis much stronger than the shoots warm. Therefore, the mostpossible period for the occurrence of sunburn of the bark ofthin branches of trees and shrubs in the result of the photodynamiceffect is March-April when the daylight hours thereare pretty significant frosts that inhibit photosynthesis andexcess light, due to an increase reflected from the snow coversolar radiation.

Isolation and characterization of a spotted leaf 32 mutant with early leaf senescence and enhanced defense response in rice

Leaf senescence is a complex biological process and defense responses play vital role for rice development, their molecular mechanisms, however, remain elusive in rice. We herein reported a rice mutant spotted leaf 32 (spl32) derived from a rice cultivar 9311 by radiation. The spl32 plants displayed early leaf senescence, identified by disintegration of chloroplasts as cellular evidence, dramatically decreased contents of chlorophyll, up-regulation of superoxide dismutase enzyme activity and malondialdehyde, as physiological characteristic, and both up-regulation of senescence-induced STAY GREEN gene and senescence-associated transcription factors, and down-regulation of photosynthesis-associated genes, as molecular indicators. Positional cloning revealed that SPL32 encodes a ferredoxin-dependent glutamate synthase (Fd-GOGAT). Compared to wild type, enzyme activity of GOGAT was significantly decreased, and free amino acid contents, particularly for glutamate and glutamine, were altered in spl32 leaves. Moreover, the mutant was subjected to uncontrolled oxidative stress due to over-produced reactive oxygen species and damaged scavenging pathways, in accordance with decreased photorespiration rate. Besides, the mutant showed higher resistance to Xanthomonas oryzae pv. Oryzae than its wild type, coupled with up-regulation of four pathogenesis-related marker genes. Taken together, our results highlight Fd-GOGAT is associated with the regulation of leaf senescence and defense responses in rice.

Distinct Mechanisms of the ORANGE Protein in Controlling Carotenoid Flux.

β-Carotene adds nutritious value and determines the color of many fruits, including melon (Cucumis melo). In melon mesocarp, β-carotene accumulation is governed by the Orange gene (CmOr) golden single-nucleotide polymorphism (SNP) through a yet to be discovered mechanism. In Arabidopsis (Arabidopsis thaliana), OR increases carotenoid levels by posttranscriptionally regulating phytoene synthase (PSY). Here, we identified a CmOr nonsense mutation (Cmor-lowβ) that lowered fruit β-carotene levels with impaired chromoplast biogenesis. Cmor-lowβ exerted a minimal effect on PSY transcripts but dramatically decreased PSY protein levels and enzymatic activity, leading to reduced carotenoid metabolic flux and accumulation. However, the golden SNP was discovered to not affect PSY protein levels and carotenoid metabolic flux in melon fruit, as shown by carotenoid and immunoblot analyses of selected melon genotypes and by using chemical pathway inhibitors. The high β-carotene accumulation in golden SNP melons was found to be due to a reduced further metabolism of β-carotene. This was revealed by genetic studies with double mutants including carotenoid isomerase (yofi), a carotenoid-isomerase nonsense mutant, which arrests the turnover of prolycopene. The yofi F2 segregants accumulated prolycopene independently of the golden SNP Moreover, Cmor-lowβ was found to inhibit chromoplast formation and chloroplast disintegration in fruits from 30 d after anthesis until ripening, suggesting that CmOr regulates the chloroplast-to-chromoplast transition. Taken together, our results demonstrate that CmOr is required to achieve PSY protein levels to maintain carotenoid biosynthesis metabolic flux but that the mechanism of the CmOr golden SNP involves an inhibited metabolism downstream of β-carotene to dramatically affect both carotenoid content and plastid fate.

Physiological differences in response to di-n-butyl phthalate (DBP) exposure between low- and high-DBP accumulating cultivars of Chinese flowering cabbage (Brassica parachinensis L.)

To increase understanding on the mechanisms of cultivar difference in contaminant accumulation in crops, this study was designed to compare the physiological responses to di-n-butyl phthalate (DBP) exposure between low (Lvbao70) and high (Huaguan) DBP cultivars of Chinese flowering cabbage (Brassica parachinensis L.). Under high DBP exposure, significant differences in various physiological responses were observed between the two cultivars, which might account for the variation in DBP accumulation. Ultrastructure observation also showed different alterations or damages in the mesophyll cell structures between both cultivars, especially for the chloroplast disintegration, starch grain quantity, and plastoglobuli accumulation. Compared with Huaguan, Lvbao70 suffered greater decreases in biomass, chlorophyll content, carbon assimilation, gas exchange parameters, photosynthetic electron transport capacity, and antioxidase activities, which would have resulted in a great reduction of photosynthetic capacity. Although Lvbao70 enhanced energy dissipation and activities of some antioxidant enzymes, they did not provide sufficient protection against oxidative damage caused by DBP. The result suggested that the lower DBP tolerance of Lvbao70 might be associated with its poor physiological performances, which was responsible for its lower DBP accumulation to protect itself from toxicity. Additionally, Lvbao70 had a significantly lower transpiration rate and stomatal conductance than Huaguan, which might be the factors regulating DBP-accumulation variation.

CYTOKININS INHIBIT LEAF SENESCENCE IN PELARGONIUM CUTTINGS

Senescence in plants is a complex, genetically determined cell death program that is characterized by protein breakdown, and disintegration of chloroplasts followed by chlorophyll loss. Cytokinins, including aromatic cytokinins and substituted phenyl urea with powerful cytokinin-like activity, are reported to have anti-senescence activities. Hence, they play an important role in delaying the onset of leaf senescence. We examined the possible use of cytokinins in delaying leaf senescence in Pelargonium A— hortorum cuttings cultivar ‘Katinka’ without adversely retarding rooting. Treated leaves maintained low leaf chroma, high hue values and high chrophyll content because they remained green. Applying aromatic cytokinin to Pelargonium cuttings had no effect on the rooting proportion (%). However, it reduced all other root parameters studied. Our study showed that the investigated cytokinins are very effective in retarding leaf senescence. When taken together with the ease of rooting cuttings, aromatic cytokinin (meta-Topolin) is a suitable alternative to phenyl urea (Thidiazuron) in delaying the onset of leaf yellowing in plants.

Selection of reference genes for qPCR- and ddPCR-based analyses of gene expression in Senescing Barley leaves.

Leaf senescence is a tightly regulated developmental or stress-induced process. It is accompanied by dramatic changes in cell metabolism and structure, eventually leading to the disintegration of chloroplasts, the breakdown of leaf proteins, internucleosomal fragmentation of nuclear DNA and ultimately cell death. In light of the global and intense reorganization of the senescing leaf transcriptome, measuring time-course gene expression patterns in this model is challenging due to the evident problems associated with selecting stable reference genes. We have used oligonucleotide microarray data to identify 181 genes with stable expression in the course of dark-induced senescence of barley leaf. From those genes, we selected 5 candidates and confirmed their invariant expression by both reverse transcription quantitative PCR and droplet digital PCR (ddPCR). We used the selected reference genes to normalize the level of the expression of the following senescence-responsive genes in ddPCR assays: SAG12, ICL, AGXT, CS and RbcS. We were thereby able to achieve a substantial reduction in the data variability. Although the use of reference genes is not considered mandatory in ddPCR assays, our results show that it is advisable in special cases, specifically those that involve the following conditions: i) a low number of repeats, ii) the detection of low-fold changes in gene expression or iii) series data comparisons (such as time-course experiments) in which large sample variation greatly affects the overall gene expression profile and biological interpretation of the data.

Post‐infectional dynamics of leaf spot disease in Withania somnifera

AbstractWithania somnifera is a high value medicinal plant, native to Indian sub‐continent, and is extensively used in traditional systems of medicine. It is highly prone to leaf spot disease caused by Alternaria alternata, that may affect the quality of crude herbal drugs. In the present study, the accumulation of ROS (reactive oxygen species) was detected histochemically within the infected host tissue, using H2DCFDA (2′,7′‐difluorodihydrofluorescein diacetate) and DAB (3,3′‐diaminobenzidine) staining methods. This ROS accumulation was linked with the process of disease initiation. The microscopy studies were conducted to explore the site of pathogen invasion into the host tissue. Some changes related to tissue degeneration during the infection process have also been reported. The observed changes in the form of reducing sugar and phenolic content showed an enhancement during infestation. Carbon, nitrogen and sulphur exhibited a decreasing trend with the progression of disease. Both the net rate of photosynthesis (A) and stomatal conductance (gS) showed a similar trend except that gS increased at the early stage of infection when the protruding fungal hyphae obstructed the stomatal closure. Further, the disintegration of chloroplasts was also observed in the infected leaves. Overall, the present study highlights a few potential targets for enhancing the disease resistance against the leaf spot pathogen of W. somnifera.

Enhanced biosynthesis of quercetin occurs as A photoprotective measure in Lycopersicon esculentum mill. under Acute UV-B exposure

Lycopersicon esculentum respond to UV-B by enhanced synthesis of flavonoid quercetin, a strong antioxidant that helps the plants to well acclimatize to UV-B stress. Three weeks old plants of L. esculentum were subjected to acute UV-B irradiation for 20, 40 and 60 minutes daily until 28 days and analyzed for the morphological and biochemical changes. UV-B exposure for 40 and 60 minutes considerably affected the growth and biomass of L. esculentum. The leaves were deformed, developed chlorosis and abscised early as compared to the unexposed plants. Biomass declined by 35% and total chlorophyll decreased by 24.7% due to disintegration of chloroplasts. Enhancement was seen in the content of carotenoids, anthocyanins and total flavonoids by 15, 33.3 and 22.8%, respectively, which was attributed to the photoprotective role of these compounds as potential quenchers of excess excitation energy. Quercetin content decreased on UV-B exposure to 20 and 40 min, and thereafter increased significantly by 5.19% on 60 min of exposure. This pattern probably indicated that the over-expression of genes involved in its biosynthesis such as phenylalanine ammonia lyase (PAL), chalcone synthase (CHS), flavanone 3-hydroxylase (F3H) and dihydroflavonol 4-reductase (DFR) occurred only after certain threshold exposure (60 min), which could be the strategy for developing tolerance against UV-B stress in L. esculentum.

Chloroplasts of Arabidopsis Are the Source and a Primary Target of a Plant-Specific Programmed Cell Death Signaling Pathway

Enhanced levels of singlet oxygen ((1)O(2)) in chloroplasts trigger programmed cell death. The impact of (1)O(2) production in chloroplasts was monitored first in the conditional fluorescent (flu) mutant of Arabidopsis thaliana that accumulates (1)O(2) upon a dark/light shift. The onset of (1)O(2) production is rapidly followed by a loss of chloroplast integrity that precedes the rupture of the central vacuole and the final collapse of the cell. Inactivation of the two plastid proteins EXECUTER (EX1) and EX2 in the flu mutant abrogates these responses, indicating that disintegration of chloroplasts is due to EX-dependent signaling rather than (1)O(2) directly. In flu seedlings, (1)O(2)-mediated cell death signaling operates as a default pathway that results in seedlings committing suicide. By contrast, EX-dependent signaling in the wild type induces the formation of microlesions without decreasing the viability of seedlings. (1)O(2)-mediated and EX-dependent loss of plastid integrity and cell death in these plants occurs only in cells containing fully developed chloroplasts. Our findings support an as yet unreported signaling role of (1)O(2) in the wild type exposed to mild light stress that invokes photoinhibition of photosystem II without causing photooxidative damage of the plant.

Regulatory Subunit B′γ of Protein Phosphatase 2A Prevents Unnecessary Defense Reactions under Low Light in Arabidopsis

Light is an important environmental factor that modulates acclimation strategies and defense responses in plants. We explored the functional role of the regulatory subunit B'γ (B'γ) of protein phosphatase 2A (PP2A) in light-dependent stress responses of Arabidopsis (Arabidopsis thaliana). The predominant form of PP2A consists of catalytic subunit C, scaffold subunit A, and highly variable regulatory subunit B, which determines the substrate specificity of PP2A holoenzymes. Mutant leaves of knockdown pp2a-b'γ plants show disintegration of chloroplasts and premature yellowing conditionally under moderate light intensity. The cell-death phenotype is accompanied by the accumulation of hydrogen peroxide through a pathway that requires CONSTITUTIVE EXPRESSION OF PR GENES5 (CPR5). Moreover, the pp2a-b'γ cpr5 double mutant additionally displays growth suppression and malformed trichomes. Similar to cpr5, the pp2a-b'γ mutant shows constitutive activation of both salicylic acid- and jasmonic acid-dependent defense pathways. In contrast to cpr5, however, pp2a-b'γ leaves do not contain increased levels of salicylic acid or jasmonic acid. Rather, the constitutive defense response associates with hypomethylation of DNA and increased levels of methionine-salvage pathway components in pp2a-b'γ leaves. We suggest that the specific B'γ subunit of PP2A is functionally connected to CPR5 and operates in the basal repression of defense responses under low irradiance.