Pigment Accumulation Research Articles

Developmental Toxicity and Teratogenic Effects of Dicarboximide Fungicide Iprodione on Zebrafish (Danio rerio) Embryos

Iprodione (IDN) is a broad-spectrum fungicide used to treat various fungal infections in plants. Despite its extensive use, assessment of its toxicity in aquatic organisms remains incomplete. This study investigated the deleterious effects of IDN using zebrafish (ZF) as a model organism. ZF embryos, beginning at 2 h post-fertilization (hpf), were exposed to IDN (3.75–40 mg/L), and both mortality and deformities were assessed. The impact of IDN on mortality was concentration-dependent and significant from 14 mg/L. Importantly, IDN induced several deformities at sublethal concentrations, including abnormal somites, reduced retinal pigment accumulation, yolk sac edema, hatching failure, abnormal swim bladders, and spinal curvature. The EC50 values for IDN-induced deformities were 3.44 ± 0.74 to 21.42 ± 6.00 mg/L. The calculated teratogenic index values for all deformities were above 1, indicating that IDN is teratogenic to ZF. IDN-exposed ZF also displayed abnormalities in touch-evoked escape responses. IDN significantly affected heart rate and blood flow, and induced pericardial edema and hyperemia in a concentration-dependent manner, suggesting its influence on cardiac development and the function of ZF. In conclusion, these results suggest that IDN exerts toxic effects on ZF embryos, affecting mortality, development, and behavior.

Revealing the effects of two DNA methyltransferase inhibitors on DNA methylation and carotenoid metabolism in Chlamydomonas reinhardtii by comparative transcriptome and physiological analysis

DNA methylation plays an important role in cell growth and development. However, little is known about the possible functions of DNA methylation in microalgae. Here, two DNA methyltransferase (DNMT) inhibitors (5-azacytidine and zebularine) were used to treat Chlamydomonas reinhardtii to investigate the effects of DNA methylation on microalgae. 5-Azacytidine (5-Azac) showed the promoting effects on cell growth and was benefit to the accumulation of pigments, while zebularine (Zeb) has adverse effects on the physiology of algal cells. However, the genomes of C. reinhardtii treated with two DNMT inhibitors unexpectedly showed slightly increased 5-mC levels. According to the results of transcriptome sequencing to algal cells treated with Zeb and 5-Azac, two DNMT inhibitors were found to up-regulate the transcription levels of genes involved in the cytosine methylation pathway and down-regulate genes involved in the cytosine demethylation pathway, which may result in higher 5-mC levels in genome. The increased content of carotenoids by 5-Azac may be due to the up-regulated transcription levels of carotenogenic genes. In addition, the up-regulated transcription levels of carotenoid degradation-related genes may be responsible for the reduced carotenoid content by Zeb. Finally, a carotenoid 9,10(9′,10′)-cleavage dioxygenase 1 (CrCCD1) gene was isolated from C. reinhardtii, and its important role in carotenoid degradation was demonstrated. In summary, this study revealed the physiological changes and molecular mechanisms of C. reinhardtii under the effects of two DNMT inhibitors, providing valuable information for subsequent studies on DNA methylation in microalgae.

Exogenous Melatonin Reinforces Photosynthesis, Antioxidant Defense and Gene Expression to Ameliorate Na2CO3 Stress in Maize.

Salt stress can seriously affect the growth and development of maize (Zea mays L.), resulting in a great yield loss. Melatonin (MT), an indole hormone, is a potential enhancer of plant tolerance against salt stress. However, the complex mechanisms of MT application in enhancing maize salt tolerance are still unclear. Herein, three-leaf seedlings of salt-susceptible P138 and its salt-resistant ethyl methane sulfonate (EMS)-104 mutant were cultured with or without 150 μM MT application under 0 and 100 mM Na2CO3 treatments for seven days, to systematically explore the response mechanisms of exogenous MT in improving the salt tolerance of maize. The results showed that salt stress triggered an escalation in reactive oxygen species production, enhanced multiple antioxidant enzymes' activities, impaired cellular membrane permeability, inhibited photosynthetic pigment accumulation, and ultimately undermined the vigor and photosynthetic prowess of the seedlings. While suitable MT application counteracted the detrimental impacts of Na2CO3 on seedlings' growth and photosynthetic capacity, the seedling length and net photosynthetic rate of P138 and EMS-104 were increased by 5.5% and 18.7%, and 12.7% and 54.5%, respectively. Quantitative real-time PCR (qRT-PCR) analysis further showed that MT application activated the expression levels of antioxidant enzyme-related genes (Zm00001d025106, Zm00001d031908, Zm00001d027511, and Zm00001d040364) and pigment biosynthesis-related genes (Zm00001d011819 and Zm00001d017766) in both maize seedlings under Na2CO3 stress; they then formed a complex interaction network of gene expression, multiple physiological metabolisms, and phenotype changes to influence the salt tolerance of maize seedlings under MT or Na2CO3 stress. To sum up, these observations underscore that 150 μM MT can alleviate salt injury of maize seedlings, which may provide new insights for further investigating MT regulation mechanisms to enhance maize seedlings' salt resistance.

Interference in Pulse Oximetry in a Neonate with Bronze Baby Syndrome.

Reliable estimation of pulse oximetry (SpO2) is indispensable in a neonatal intensive care setting. Along with clinical examination, SpO2 is a validated non-invasive correlate of PaO2 to dictate the level of respiratory support. Any alterations that may interfere with this correlation pose significant challenges and lead to inadvertent over- or under-treatment. The authors report a case with an unusual discrepancy between SpO2 and PaO2 in a neonate with jaundice who developed bronze baby syndrome while receiving phototherapy. A review of similar previous reports suggests that abnormal pigment accumulation in bronze baby syndrome may render pulse oximetry transiently unreliable.

Alleviation of NaCl Stress on Growth and Biochemical Traits of Cenchrus ciliaris L. via Arbuscular Mycorrhizal Fungi Symbiosis

Soil salinization, especially in arid and semi-arid regions, is one of the major abiotic stresses that affect plant growth. To mediate and boost plant tolerance against this abiotic stress, arbuscular mycorrhizal fungi (AMF) symbiosis is commonly thought to be an effective tool. So, the main purpose of this study was to estimate the role of AMF (applied as a consortium of Claroideoglomus etunicatum Claroideoglomus eutinicatum, Funneliformis mosseae Funliformis mosseae, Rhizophagus fasciculatum, and R. intraradices species) symbiosis in mitigating deleterious salt stress effects on the growth parameters (shoot length (SL), root length (RL), shoot dry weight (SDW), root dry weight (RDW), root surface area (RSA), total root length (TRL), root volume (RV), root diameter (RD), number of nodes and leaves) of Cenchrus ciliaris L. plants through improved accumulations of photosynthetic pigments (chlorophyll a, chlorophyll b, total chlorophyll), proline and phenolic compounds. The results of this experiment revealed that the roots of C. ciliaris plants were colonized by AMF under all the applied salinity levels (0, 75, 150, 225, and 300 mM NaCl). However, the rate of colonization was negatively affected by increasing salinity as depicted by the varied colonization structures (mycelium, vesicles, arbuscules and spores) which were highest under non-saline conditions. This association of AMF induced an increase in the growth parameters of the plant which were reduced by salinity stress. The improved shoot/root indices are likely due to enhanced photosynthetic activities as the AMF-treated plants showed increased accumulation of pigments (chlorophyll a, chlorophyll b and total chlorophyll), under saline as well as non-saline conditions, compared to non-AMF (N-AMF) plants. Furthermore, the AMF-treated plants also exhibited enhanced accumulation of proline and phenolic compounds. These accumulated metabolites act as protective measures under salinity stress, hence explaining the improved photosynthetic and growth parameters of the plants. These results suggest that AMF could be a good tool for the restoration of salt-affected habitats. However, more research is needed to check the true efficacy of different AMF inoculants under field conditions.

Integrated Metabolomics and Transcriptomics Analyses Reveal the Regulatory Mechanisms of Anthocyanin and Carotenoid Accumulation in the Peel of Coffea arabica.

The color of coffee fruits is influenced by several factors, including cultivar, ripening stage, and metabolite composition. However, the metabolic accumulation of pigments and the molecular mechanisms underlying peel coloration during the ripening process of Coffea arabica L. remain relatively understudied. In this study, UPLC-MS/MS-based metabolomics and RNA sequencing (RNA-seq)-based transcriptomics were integrated to investigate the accumulation of anthocyanins and carotenoids in the peel of Coffea arabica at different ripening stages: green peel (GP), green-yellow peel (GYRP), red peel (RP), and red-purple peel (RPP). This integration aimed at elucidating the molecular mechanisms associated with these changes. A total of ten anthocyanins, six carotenoids, and thirty-five xanthophylls were identified throughout the ripening process. The results demonstrated a gradual decrease in the total carotenoid content in the peel with fruit maturation, while anthocyanin content increased significantly. Notably, the accumulation of specific anthocyanins was closely associated with the transition of peel colors from green to red. Integrated metabolomics and transcriptomics analyses identified the GYRP stage as critical for this color transition. A weighted gene co-expression network analysis (WGCNA) revealed that enzyme-coding genes such as 3AT, BZ1, and lcyE, along with transcription factors including MYB, NAC, and bHLH, which interact with PHD and SET TR, may regulate the biosynthesis of anthocyanins and carotenoids, thereby influencing peel pigmentation. These findings provide valuable insights into the molecular mechanisms underlying the accumulation of anthocyanins and carotenoids in Coffea arabica peel during fruit maturation.

Optimization of iron, phosphate, and salinity in nutrient medium using response surface methodology for enhancing biochemical composition in Chlorella sp. culture

Microalga Chlorella sp. is a highly scientifically and commercially attractive unicellular microorganism and has developed a stable industry as a nutritional supplement for people and animals. Various nutrient media have been used to grow Chlorella sp. cultures to enhance their growth, pigmentation, and lipid content. However, the optimal biochemical composition and density of Chlorella sp. cultures require an understanding of optimizing the nutrient medium to study their production. The present study aims to investigate the effects of iron, phosphate, and salinity concentrations by working synergically in a nutrient medium on the growth responses, pigments, and lipid accumulation of Chlorella sp. culture using a response surface methodology (RSM) approach. Using the result from the RSM software, a total of 18 experimental groups (E1 – E18) were evaluated, and one confirmation (C) study was conducted. The results revealed that the E7 experiment in Chlorella sp. culture provided the highest cell density and specific growth rate (SGR) with 176.00 × 106 cells mL−1 and 0.35 day−1, respectively. Similarly, the E5 and E1 experiments produced the highest cell density of 166.10 × 106 cells. mL−1 and 152.13 × 106 cells mL−1, respectively. The SGR was also increased at 0.33 day−1 in the E5 experiment and 0.33 day−1 in the E1 experiment. Consequently, the culture of Chlorella sp. containing high iron and phosphate concentrations and lower salinity in a nutrient medium had the highest number of cells, SGR, and pigment accumulation (chlorophyll a and total carotenoid). In addition, the presence of high salinity concentrations reduces Chlorella sp. growth. However, the increase in the growth of Chlorella sp. culture did not indicate an increase in other biochemical compositions. In some cases, biochemical compositions are high due to nutritional limitations or stress factors. For example, in pigment accumulation, chlorophyll a pigment accumulation was increased in experiment E7 (51.57 μg mL−1), while total carotenoid accumulation was increased in experiment E8 (20.68 μg mL−1). In addition, increasing salinity concentration increased chlorophyll a and total carotenoid contents per cell, but decreased Chlorella sp. growth as shown in the E4 experiment, which achieved chlorophyll a levels of 1.38 pg cell−1 and total carotenoid levels of 0.52 pg cell−1. Furthermore, Chlorella sp. culture produces a higher lipid accumulation of 37.38 % in the E3 experiment. Hence, the results of this study contribute to understanding the optimal biochemical composition and cell growth of Chlorella sp. cultures.

An in vitro cell model for exploring inflammatory and amyloidogenic events in alkaptonuria.

Alkaptonuria (AKU) is a progressive systemic inherited metabolic disorder primarily affecting the osteoarticular system, characterized by the degeneration of cartilage induced by ochronosis, ultimately leading to early osteoarthritis (OA). However, investigating AKU pathology in human chondrocytes, which is crucial for understanding the disease, encounters challenges due to limited availability and donor variability. To overcome this obstacle, an in vitro model has been established using homogentisic acid (HGA) to simulate AKU conditions. This model employed immortalized C20/A4 human chondrocytes and serves as a dependable platform for studying AKU pathogenesis. Significantly, the model demonstrates the accumulation of ochronotic pigment in HGA-treated cells, consistent with findings from previous studies. Furthermore, investigations into inflammatory processes during HGA exposure revealed notable oxidative stress, as indicated by elevated levels of reactive oxygen species and lipid peroxidation. Additionally, the model demonstrated HGA-induced inflammatory responses, evidenced by increased production of nitric oxide, overexpression of inducible nitric oxide synthase, and cyclooxygenase-2. These findings underscore the model's utility in studying inflammation associated with AKU. Moreover, analysis of serum amyloid A and serum amyloid P proteins revealed a potential interaction, corroborating evidence of amyloid fibril formation. This hypothesis was further supported by Congo red staining, which showed fibril formation exclusively in HGA-treated cells. Overall, the C20/A4 cell model provided valuable insights into AKU pathogenesis, emphasizing its potential for facilitating drug development and therapeutic interventions.

UV-B light (radiation) affects the metabolism of pigments and fatty acids in green algae Edaphochlorella mirabilis and Klebsormidium flaccidum in vitro

Algae offer a rich source of bioactive compounds suitable for food products and bioenergy. Environmental challenges such as nutrient scarcity, extreme pH and temperature, high light intensity, and UV radiation usually trigger algae to produce excess of lipids, antioxidants, and other bioprotective molecules as part of their adaptations for survival. Algal cultivation provides proteins, lipids, carbohydrates, vitamins, antioxidants, and trace elements. This study focused on understanding how UV-B irradiance, as an abiotic stressor, can influence the growth and metabolite production of two green algal species, Edaphochlorella mirabilis (Chlorophyta) and Klebsormidium flaccidum (Charophyta). Using a temporary immersion system bioreactor for in-vitro algal growth, the results showed no significant difference in biomass for both algal species after the exposure to UV-B rays. However, the assessment of malondialdehyde levels revealed a significantly higher tendency towards lipid peroxidation in treated E. mirabilis (+ 90 %) compared to control. Conversely, K. flaccidum did not display significant differences, thereby highlighting a more advanced adaptive capacity against UV-B radiation. Overall, both algal species treated with UV-B showed increased pigment accumulation. K. flaccidum exhibited an average pigment increase of over 53 %, while E. mirabilis showed a lower increase, over 30 % on average. The notable rise in antioxidant compounds (lutein, β-carotene, and chlorophyll a) in UV-B exposed K. flaccidum samples also suggested a more suitable adaptive strategy to mitigate oxidative stress in Charophyta. In K. flaccidum, the increase in polyunsaturated fatty acids can be associated with increased production of antioxidant compounds. Conversely, E. mirabilis appeared to protect itself by decreasing polyunsaturated fatty acids in favor of saturated ones. In both algal species, the increase in secondary metabolites under UV stress highlighted potential as a novel food source for human consumption, deserving further investigation.

Low temperature and high humidity affect dynamics of chlorophyll biosynthesis and secondary metabolites in Cucumber.

During the cold season, low temperature (LT) and high relative humidity (HRH) are significant environmental factors in greenhouses and plastic tunnels, often hindering plant growth and development. The chlorophyll (Chl) biosynthesis inhibitory mechanisms under LT and HRH stress are still widely unclear. To understand how cucumbers seedlings respond to LT and HRH stress, we investigated the impact of these stressors on Chl biosynthesis. Our results revealed that individual LT, HRH and combined LT + HRH stress conditions affected chlorophyll a, b, total chlorophyll and carotenoid content, reducing the levels of these pigments. The levels of Chlorophyll precursors were also markedly reduced under LT and HRH stresses, with the greatest reduction observed in cucumber seedlings exposed to LT + HRH conditions (9/5℃, 95%HRH). The activities of glutamate-1-semialdehyde transaminase (GSA-AT), ALA dehydratase (ALAD), Mg-chelatase, and protochlorophyllide oxidoreductase (POR) were increased under individual LT, HRH, conditions but decreased by combination of LT + HRH stress condition. In addition, Chl biosynthesis related genes (except PBG) were upregulated by the HRH stress but were significantly downregulated under the LT + HRH stress condition in cucumber seedlings. Furthermore, the content of phenols, flavonoids and phenolic acids (cinnamic acid and caffeic acid) were significantly surged under LT + HRH treatment over the control. Histochemical observation showed higher O2- and H2O2 content in cucumber leaves during the LT and HRH stress. The results indicate that LT + HRH stress significantly impairs chlorophyll biosynthesis in cucumber seedlings by drastically reducing pigment accumulation, altering enzyme activity and gene expression. Additionally, LT + HRH stress induces oxidative damage, which further exacerbates the decline in chlorophyll content and affects overall cucumber metabolism.

Influence of Primary Light Exposure on the Morphophysiological Characteristics and Phenolic Compounds Accumulation of a Tea Callus Culture (Camellia sinensis L.).

Tea plant calli (Camellia sinensis L.) are characterized by the accumulation of various phenolic compounds (PC)-substances with high antioxidant activity. However, there is still no clarity on the response of tea cells to light exposure of varying intensity. The purpose of the research was to study tea callus cultures grown under the influence of primary exposure to different light intensities (50, 75, and 100 µmol·m-2·s-1). The cultures' growth, morphology, content of malondialdehyde and photosynthetic pigments (chlorophyll a and b), accumulation of various PC, including phenylpropanoids and flavanols, and the composition of catechins were analyzed. Primary exposure to different light intensities led to the formation of chloroplasts in tea calli, which was more pronounced at 100 µmol·m-2·s-1. Significant similarity in the growth dynamics of cultures, accumulation of pigments, and content of malondialdehyde and various phenolics in tea calli grown at light intensities of 50 and 75 µmol·m-2·s-1 has been established, which is not typical for calli grown at 100 µmol·m-2·s-1. According to data collected using high-performance liquid chromatography, (+)-catechin, (-)-epicatechin, epigallocatechin, gallocatechin gallate, epicatechin gallate, and epigallocatechin gallate were the main components of the tea callus culture's phenolic complex. Its content changed under the influence of primary exposure to light, reaching the greatest accumulation in the final stages of growth, and depended on the light intensity. The data obtained indicate changes in the morphophysiological and biochemical characteristics of tea callus cultures, including the accumulation of PC and their individual representatives under primary exposure to light exposure of varying intensity, which is most pronounced at its highest values (100 µmol·m-2·s-1).

The auronidin flavonoid pigments of the liverwort Marchantia polymorpha form polymers that modify cell wall properties.

Plant adaptation from aquatic to terrestrial environments required modifications to cell wall structure and function to provide tolerance to new abiotic and biotic stressors. Here, we investigate the nature and function of red auronidin pigment accumulation in the cell wall of the liverwort Marchantia polymorpha. Transgenic plants with auronidin production either constitutive or absent were analysed for their cell wall properties, including fractionation of polysaccharide and phenolic components. While small amounts of auronidin and other flavonoids were loosely associated with the cell wall, the majority of the pigments were tightly associated, similar to what is observed in angiosperms for polyphenolics such as lignin. No evidence of covalent binding to a polysaccharide component was found: we propose auronidin is present in the wall as a physically entrapped large molecular weight polymer. The results suggested auronidin is a dual function molecule that can both screen excess light and increase wall strength, hydrophobicity and resistance to enzymatic degradation by pathogens. Thus, liverworts have expanded the core phenylpropanoid toolkit that was present in the ancestor of all land plants, to deliver a lineage-specific solution to some of the environmental stresses faced from a terrestrial lifestyle.

Accumulation of plastid pigments in the leaves Triticum avestium L. under influence of composite recultivant Trevitan®

Aim. To investigate the efficiency of using composite recultivants TREVITAN® (RCT) according to the parameters of accumulation of the photosynthetic pigments in the leaves Triticum aestivum L. of the varieties Quintus and Licamero. Methods. The experiments were carried out on the plots of the agro-biolaboratory of Ternopil Volodymyr Hnatyuk National Pedagogical University in 4 variants (Control, TREVITAN® adjuvant, recultivants TREVITAN®, TREVITAN® adjuvant + recultivants TREVITAN®) and 3 repetitions. The content of chlorophylls a, b and carotenoids in flag leaves was defined by Welburn spectrometric method. Results. The highest content of chlorophylls was found in the leaves of both varieties of wheat in the phases of flowering and milk ripeness during autumn tillage, seeds before sowing and plants in the earing phase with recultivants TREVITAN®. Joint use of TREVITAN® adjuvant + recultivants TREVITAN® also significantly increased the content of chlorophyll in wheat leaves. Increase of carotenoids in the leaves of plants of all experimental variants on 4.2–14.6 % (variety Quintus) and on 43.2–59.5 % (variety Licamero) was defined in the phase of milk ripeness. Conclusions. The use of RCT in the technology of growing soft wheat had a significant impact on the accumulation of photosynthetic pigments in leaves during the generative phases of development.

Virus-Induced galactinol-sucrose galactosyltransferase 2 Silencing Delays Tomato Fruit Ripening.

Tomato fruit ripening is an elaborate genetic trait correlating with significant changes at physiological and biochemical levels. Sugar metabolism plays an important role in this highly orchestrated process and ultimately determines the quality and nutritional value of fruit. However, the mode of molecular regulation is not well understood. Galactinoal-sucrose galactosyltransferase (GSGT), a key enzyme in the biosynthesis of raffinose family oligosaccharides (RFOs), can transfer the galactose unit from 1-α-D-galactosyl-myo-inositol to sucrose and yield raffinose, or catalyze the reverse reaction. In the present study, the expression of SlGSGT2 was decreased by Potato Virus X (PVX)-mediated gene silencing, which led to an unripe phenotype in tomato fruit. The physiological and biochemical changes induced by SlGSGT2 silencing suggested that the process of fruit ripening was delayed as well. SlGSGT2 silencing also led to significant changes in gene expression levels associated with ethylene production, pigment accumulation, and ripening-associated transcription factors (TFs). In addition, the interaction between SlGSGT2 and SlSPL-CNR indicated a possible regulatory mechanism via ripening-related TFs. These findings would contribute to illustrating the biological functions of GSGT2 in tomato fruit ripening and quality forming.

Exogenous Uniconazole promotes physiological metabolism and grain yield of rice under salt stress.

Salt stress severely inhibit plant growth and development. Uniconazole has been considered to significantly increase plant stress tolerance. However, the mechanism by which Uniconazole induces salt tolerance in rice seedlings and its impact on yield is still unclear. In this study, the effects of exogenous Uniconazole on morphogenesis, physiological metabolism, and yield of rice seedlings under salt stress were analyzed using the salt-tolerant rice variety HD961 and the salt-sensitive rice variety 9311. The results showed that salt stress significantly inhibited rice growth, disrupted the antioxidant system and pigment accumulation, and reduced photosynthesis, and yield. There were corresponding percent decreases of 13.0% and 24.1% in plant height, 31.6% and 55.8% in leaf area, 65.7% and 85.3% in root volume, respectively for HD961 and 9311. spraying However, compared to salt stress, the US treatment increased the percentage to 4.7% and 139.0% in root volume, 7.5% and 38.0% in total chlorophyll, 4.5% and 14.3% in peroxidase (POD) of leaves, 14.4% and 54.2% in POD of roots, 18.7% and 22.7% in catalase (CAT) of leaves, and 22.6% and 53.9% in CAT of roots, respectively, for HD961 and 9311. In addition, it also significantly enhanced photosynthesis at the reproductive stage, promoted the transport of carbohydrate to grains. And US treatment significantly increased the percentage to 9.0% in panicle length, 28.0% in panicle number per hole, 24.0% in filled grain number, 3.0% in 1000-grain weight, and 26.0% in yield per plant, respectively, for HD961, compared to salt stress. In summary, applying Uniconazole at the seedling stage can alleviate the damage induced by NaCl stress on rice by regulating the physiological metabolism of rice plants. This reduces the negative effects of salt stress, enhance salt tolerance, and boost rice production.

Functional variations in efficiency of PSII during leaf ontogeny in the tropical plant Saraca asoca

Leaf ontogeny of tropical evergreen tree species lasts several months with changes in size, shape, colouration and internal tissue distribution of leaves. Leaf initiation in Saraca asoca generally occurs once in a year during February–April, followed by very limited leafing thereafter. We measured the rate of photosynthesis, chlorophyll a fluorescence, energy quenching and PSII functions during the leaf ontogeny process. Observations were taken up to 35 days after opening of lamina (DAOL). Significant increase in the synthesis and accumulation of photosynthetic pigments but negative net photosynthesis was noticed during initial days of the ontogeny. The leaf moved from heterotrophy to autotrophy with gradual improvement of PSII functions. The ratio of intercellular CO2 (Ci) and ambient CO2 (Ca) showed significant change at ≥11 DAOL. Increase in the age of the leaf (between 5 and 28 DAOL) caused decrease in O-J rise and corresponding increase in J-I and I-P rise as well as of fluorescence maximum (FM) of the OJIP curve. The improvement of the electron transport components of the donor side of PSII was seen with increase in the functional oxygen evolving complex. The functional improvements of the donor and acceptor side of PSII during leaf ontogeny are discussed.

Ocular melanosis in the adult Cairn Terrier population within the United Kingdom.

Prospective investigation to determine the prevalence of ocular melanosis in adult Cairn Terriers within the United Kingdom using a previously established staging scheme. Ophthalmic assessment was performed on adult Cairn Terriers, recruited from various geographic locations within the United Kingdom. Examination included gonioscopy, rebound tonometry, slit lamp biomicroscopy, and indirect ophthalmoscopy, performed by one examiner (AM). A total of 93 dogs were examined, including 52 females and 41 males, aged between 15 months and 16 years 4 months. Sixty of 93 dogs (64.5%) were >7 years of age. Nine of 93 dogs (9.6%) demonstrated changes consistent with ocular melanosis. Four of 9 (44.4%) had Stage 1 disease and 5 of 9 (55.6%), Stage 2. Stages 3 or 4 were not identified in any dogs. Mean intraocular pressures in affected and unaffected dogs were 14.7 mmHg (range 12-17 mmHg) and 12.8 mmHg (range 5-21 mmHg), respectively. Incomplete pupil dilation was noted in affected dogs following pharmacologic mydriasis. Ocular melanosis was identified in approximately 10% of examined dogs, over half were dogs of breeding age (<7 years of age). It is possible that Grade 1 disease could go undetected, prior to obvious scleral pigment accumulation (Grade 2 disease). It is therefore recommended that dogs undergoing pre-breeding screens have pre-dilation assessment of the anterior segment using slit lamp biomicroscopy with subsequent gonioscopy to clearly assess for circumferential thickening of the iris base that might otherwise go undetected. Additionally, regular reassessment of breeding dogs is advised as disease progression could be rapid.

Genome-Wide Identification of B-Box Family Genes and Their Potential Roles in Seed Development under Shading Conditions in Rapeseed.

B-box (BBX) proteins, a subfamily of zinc-finger transcription factors, are involved in various environmental signaling pathways. In this study, we conducted a comprehensive analysis of BBX family members in Brassica crops. The 482 BBX proteins were divided into five groups based on gene structure, conserved domains, and phylogenetic analysis. An analysis of nonsynonymous substitutions and (Ka)/synonymous substitutions (Ks) revealed that most BBX genes have undergone purifying selection during evolution. An analysis of transcriptome data from rapeseed (Brassica napus) organs suggested that BnaBBX3d might be involved in the development of floral tissue-specific RNA-seq expression. We identified numerous light-responsive elements in the promoter regions of BnaBBX genes, which were suggestive of participation in light signaling pathways. Transcriptomic analysis under shade treatment revealed 77 BnaBBX genes with significant changes in expression before and after shading treatment. Of these, BnaBBX22e showed distinct expression patterns in yellow- vs. black-seeded materials in response to shading. UPLC-HESI-MS/MS analysis revealed that shading influences the accumulation of 54 metabolites, with light response BnaBBX22f expression correlating with the accumulation of the flavonoid metabolites M46 and M51. Additionally, BnaBBX22e and BnaBBX22f interact with BnaA10.HY5. These results suggest that BnaBBXs might function in light-induced pigment accumulation. Overall, our findings elucidate the characteristics of BBX proteins in six Brassica species and reveal a possible connection between light and seed coat color, laying the foundation for further exploring the roles of BnaBBX genes in seed development.

Inhibitory Effects, Fluorescence Studies, and Molecular Docking Analysis of Some Novel Pyridine-Based Compounds on Mushroom Tyrosinase.

Melanin biosynthesis in different organisms is performed by a tyrosinase action. Excessive enzyme activity and pigment accumulation result in different diseases and disorders including skin cancers, blemishes, and darkening. In fruits and vegetables, it causes unwanted browning of these products and reduces their appearance quality and economic value. Inhibiting enzyme activity and finding novel powerful and safe inhibitors are highly important in agriculture, food, medical, and pharmaceutical industries. In this regard, in the present study, some novel synthetic pyridine-based compounds including 2,6-bis (tosyloxymethyl) pyridine (compound 3), 2,6-bis (butylthiomethyl) pyridine (compound 4), and 2,6-bis (phenylthiomethyl) pyridine (compound 5) were synthesized for the first time, and their inhibitory potencies were assessed on mushroom tyrosinase diphenolase activity. The results showed that while all tested compounds significantly decreased the enzyme activity, compounds 4 and 5 had the highest inhibitory effects (respectively, 80 and 89% inhibition with the IC50 values of 17.0 and 9.0 μmol L-1), and the inhibition mechanism was mixed-type for both compounds. Ligand-binding studies were carried out by fluorescence quenching and molecular docking methods to investigate the enzyme-compound interactions. Fluorescence quenching results revealed that the compounds can form nonfluorescent complexes with the enzyme and result in quenching of its intrinsic emission by the static process. Molecular docking analyses predicted the binding positions and the amino acid residues involved in the interactions. These compounds appear to be suitable candidates for more studies on tyrosinase inhibition.

Tyrosinase-induced neuromelanin accumulation triggers rapid dysregulation and degeneration of the mouse locus coeruleus.

The locus coeruleus (LC), the major source of norepinephrine (NE) in the brain, is an early site of pathology in both Alzheimer's disease (AD) and Parkinson's disease (PD), and it undergoes catastrophic degeneration later in both disorders. Dysregulation of the LC is thought to contribute to prodromal symptoms of AD and PD such as anxiety and sleep disturbances, while frank LC-NE loss promotes cognitive decline. However, the mechanisms responsible for its selective vulnerability are unknown. The LC is among the only structures in the brain that produces appreciable amounts of neuromelanin (NM), a dark cytoplasmic pigment. It has been proposed that NM initially plays a protective role by sequestering toxic catecholamine metabolites and heavy metals, but may become harmful during aging as it overwhelms cellular machinery and is released during neurodegeneration. Rodents do not naturally produce NM, limiting the study of causal relationships between NM and LC pathology. Adapting a viral-mediated approach for expression of human tyrosinase, the enzyme responsible for peripheral melanin production, we successfully promoted pigmentation in mouse LC neurons that recapitulates key ultrastructural features of endogenous NM found in primates. Pigment expression results in LC neuron hyperactivity, reduced tissue NE levels, transcriptional changes, and novelty-induced anxiety phenotypes as early as 1-week post-injection. By 6-10 weeks, NM accumulation is associated with severe LC neuron neurodegeneration and microglial engulfment of the pigment granules, while the anxiety-like behavior is abated. These phenotypes are reminiscent of LC dysfunction and cell death in AD and PD, validating this model for studying the consequences of pigment accumulation in the LC as it relates to neurodegenerative disease.