Higher Superoxide Dismutase Activity Research Articles

Effect of heat acclimation on thermotolerance of in vitro strawberry plantlets

ABSTRACT Strawberry production is facing a serious decline with the increase in global temperature as a result of climate change. Improving the heat tolerance is imperative for the strawberry plants to remain productive under high temperature conditions. The present work aimed to study the effect of heat acclimation on the thermotolerance of strawberry plants subjected to severe heat stress. Tissue cultured Fragaria ‘Taoyuan No. 1’ plantlets were subjected to four heat-acclimation treatments with gradual increase of temperatures from 30°C to 42°C for 1.25 hr to 10 hr before exposing them to the lethal temperature of 48°C for 4 hr. Survival, new leaf emergence and root growth, electrolyte leakage, chlorophyll and proline contents, as well as antioxidant enzyme activities were compared between the control, acclimated and non-acclimated plantlets. Results indicated that heat acclimation was required for the strawberry plantlets to survive under lethal temperature conditions. The acclimated plantlets registered a lower degree of electrolyte leakage and chlorophyll degradation, and a higher proline content compared to the non-acclimated plantlets. The activities of the antioxidant enzymes increased with the elevation of acclimation temperature and peaked at 42°C except for ascorbate peroxidase (APX) whose activity peaked at 39°C. Higher activities of superoxide dismutase (SOD), APX, glutathione reductase (GR), catalase (CAT) and peroxidase (POD) were observed in the acclimated plantlets compared to the non-acclimated plantlets. This study demonstrates that heat acclimation improved the thermotolerance of in vitro strawberry plantlets by reducing electrolyte leakage and chlorophyll degradation, as well as by enhancing proline content and antioxidant enzyme activities under severe heat stress.

Physiological Regulation of Photosynthetic-Related Indices, Antioxidant Defense, and Proline Anabolism on Drought Tolerance of Wild Soybean (Glycine soja L.)

Wild soybean (Glycine soja L.), drought-tolerant cultivar Tiefeng 31 (Glycine max L.), and drought-sensitive cultivar Fendou 93 (Glycine max L.) were used as materials to investigate the drought tolerance mechanism after 72 h 2.5 M PEG 8000 (osmotic potential −0.54 MPa)-simulated drought stress at the seedling stage. The results indicated that the leaves of the G. soja did not wilt under drought stress. However, both the drought-tolerant and drought-sensitive cultivated soybean cultivars experienced varying degrees of leaf wilt. Notably, the drought-sensitive cultivated soybean cultivars exhibited severe leaf wilt after the drought stress. Drought stress was determined to have a significant impact on the dry matter of the above-ground part of the drought-sensitive cultivar Fendou 93, followed by the drought-tolerant cultivar Tiefeng 31, with the lowest reduction observed in G. soja. Furthermore, the presence of drought stress resulted in the closure of leaf stomata. G. soja exhibited the highest proportion of stomatal opening per unit area, followed by the drought-tolerant cultivar Tiefeng 31, while the drought-sensitive cultivar Fendou 93 displayed the lowest percentage. Photosynthesis-related indexes, including photosynthetic rate, intercellular CO2, transpiration rate, and stomatal conductance, decreased in Fendou 93 and Tiefeng 31 after drought stress, but increased in G. soja. In terms of the antioxidant scavenging system, lower accumulation of malondialdehyde (MDA) was observed in G. soja and Tiefeng 31, along with higher activities of superoxide dismutase (SOD, EC 1.15.1.1) and catalase (CAT, EC 1.11.1.6) to counteract excess reactive oxygen species and maintain cell membrane integrity. In contrast, the drought-sensitive cultivar Fendou 93 had higher MDA content and higher activities of ascorbate peroxidase (APX, EC 1.11.1.11) and peroxidase (POD, 1.11.1.7). G. soja and Tiefeng 31 also exhibited less accumulation of osmolytes, including soluble sugar, soluble protein, and free proline content. The activities of δ-OAT, ProDH, and P5CS, key enzymes in proline anabolism, showed an initial increase under drought stress, followed by a decrease, and then an increase again at the end of drought stress in G. soja. Before drought stress, Tiefeng 31 had higher activities of ProDH and P5CS, which decreased with prolonged drought stress. Fendou 93 experienced an increase in the activities of δ-OAT, ProDH, and P5CS under drought stress. The δ-OAT gene expression levels were up-regulated in all three germplasms. The expression levels of the P5CS gene in Fendou 93 and Tiefeng 31 were down-regulated, while G. soja showed no significant change. The expression of the P5CR gene and ProDH gene was down-regulated in Fendou 93 and Tiefeng 31, but up-regulated in G. soja. This indicates that proline content is regulated at both the transcription and translation levels.

The Effect of Grape Seed Cake as a Dietary Supplement Rich in Polyphenols on the Quantity and Quality of Milk, Metabolic Profile of Blood, and Antioxidative Status of Lactating Dairy Goats

The objective of this study was to assess the impact that diets supplemented with grape seed cake rich in polyphenols had on lactating goats. The study investigated the quantity and quality of goat milk, the metabolic profile of blood, and the antioxidative status. The study involved 24 French Alpine dairy goats throughout their lactation period. The goats were, on average, 5 years old (±three months) and in the fourth lactation. The experiment lasted for 58 days. The control group (CON) had a diet without grape seed cake (GSC). The experimental groups were given a diet containing 5% and 10% GSC on a dry matter basis (GSC5 and GSC10, respectively). A slightly higher milk production, as well as protein and fat milk content, were found in GSC5 and GSC10, but the differences were not significant. Goat milk in the GSC10 group exhibited significantly higher activity of superoxide dismutase and glutathione reductase, as well as decreased concentrations of GUK and SCC. The feeding treatments did not affect significant differences in hematological and biochemical indicators, except for the BHB content, which can be associated with a higher energy value of feed containing GSC. There was an observed elevation in the activity of SOD within the blood of GSC5, and GSC10 was measured as well. The determined changes justify the supplementation of GSC rich in polyphenols to goat feed, especially in the amount of 10%, as it can reduce stress caused by lactation, which is known as a very stressful production period for animals.

Elevation-driven modifications in tissue architecture and physiobiochemical traits of Panicum antidotale Retz. in the Pothohar Plateau, Pakistan

Mountainous ecosystems are experiencing rapid shifts in distribution patterns and community structure due to the impacts of climate change and vegetation dynamics. This study offers a comprehensive insight into the adaptive mechanisms of Panicum antidotale to varying elevations within the Pothohar Plateau, shedding light on its survival strategies in climate-affected regions. The findings reveal that populations at lower elevations exhibit maximum height, leaf area, shoot length, and fresh weight, whereas those at higher elevations display significant decrease in these parameters. Moreover, a significant decline in species density, frequency and cover is evident at high elevation. High-elevation populations demonstrate decreased photosynthetic traits, including chlorophyll, carotenoid, and Na and Ca content. Furthermore, there are notable variations in the antioxidant enzyme activity across elevation gradients. Populations at lower elevations exhibit higher activities of superoxide dismutase and peroxidase. Across the gradient, there were significant reductions in root and stem radius, sclerenchyma thickness, bulliform cell size, metaxylem vessels, and phloem area. Populations at moderate elevations displayed thicker lamina and epidermis, larger cortical cells, wider xylem vessels, and increased phloem area; however, these traits declined at higher elevations. High-elevation populations showed thicker lamina and epidermis, extensive sclerenchyma, greater cortical cell area, and higher stomatal density. Photosynthetic traits and enzymatic activity were major contributors at moderate elevations while stem and leaf traits were significantly changed at high elevations. Overall, this research contributes significantly to the broader knowledge of plant adaptation in mountainous ecosystems and stresses the importance of considering elevation as a critical factor in ecological studies.

The effect of nettle extract on antioxidant defense system in piglets after weaning

Background. The effect of common nettle (Urtica dioica L.) extracts on the free radical processes and antioxidant system in piglets during the critical period of weaning from sows has been studied. Materials and Methods. Large white piglets were divided into 2 groups (control and experimental), 9 animals in each. Piglets of the experimental group from 14 days of age and before weaning received the standard diet and the nettle extract in the dose of 6 mg/kg of body weight for 22 days. The blood, as well as erythrocyte hemolysates and plasma of piglets obtained at 14, 36, and 42 days of age, were studied. Results. Our results have shown that weaning causes an oxidative stress in piglets. This state leads to an increase in the concentration of metabolites of free radical damage to protein molecules – carbonyl groups of proteins on the first day and primary products of lipid peroxidation on the seventh day after weaning. This activation of oxidative damage occurs in piglets against the background of a physiologically immature antioxidant system and is characterized by a decrease in the activity of the enzymatic chain – superoxide dismutase, glutathione peroxidase and catalase, as well as the concentration of its non-enzymatic antioxidant – reduced glutathione. Feeding piglets with nettle extract leads to activation of the antioxidant defense system in erythrocytes (higher activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and an increased reduced glutathione level compared to the control values) and a decrease in the concentration of oxidative damage products in the plasma (the content of lipid hydroperoxides, TBA-active products and carbonyl groups of proteins). Conclusion. The positive effect of nettle extract on the inhibition of free radical processes and activation of antioxidant systems indicates that this extract can be added to the standard diet of young animals to increase stress resistance and adaptability of their organism in critical periods of ontogenesis.

Topless–related 2 conferred cadmium accumulation in wheat

Wheat is a vital food crop that faces threats from various abiotic and biotic stresses. Understanding the molecular mechanism of cadmium (Cd) resistance can provide valuable insights into the tolerance of wheat. Plant proteins known as Topless/Topless–Related (TPL/TPR) play a role in growth, development, defense regulation, and stress response. In this study, we identified TaTPR2 as being induced by Cd stress treatment. Upon Cd treatment, wheat plants overexpressing TaTPR2 exhibited better growth compared to wild–type (WT) plants. Moreover, the transgenic lines showed reduced accumulation of reactive oxygen species (ROS), along with significantly higher activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) compared to WT plants. Additionally, the transgenic lines exhibited lower levels of malondialdehyde (MDA) and electrolyte leakage compared to WT plants. Further analysis revealed that TabHLH41 directly binds to the E–box motif of the TaTPR2 promoter and positively regulates its expression. Overall, the overexpression of TaTPR2 in transgenic wheat resulted in reduced accumulation of Cd and ROS. These findings highlight the significance of the TabHLH41–TaTPR2 pathway as a crucial response to Cd stress in wheat.

[Effects of cerium oxide nanoenzyme-gelatin methacrylate anhydride hydrogel in the repair of infected full-thickness skin defect wounds in mice].

Objective: To investigate the effects of cerium oxide nanoenzyme-gelatin methacrylate anhydride (GelMA) hydrogel (hereinafter referred to as composite hydrogel) in the repair of infected full-thickness skin defect wounds in mice. Methods: This study was an experimental study. Cerium oxide nanoenzyme with a particle size of (116±9) nm was prepared by hydrothermal method, and GelMA hydrogel with porous network structure and good gelling performance was also prepared. The 25 μg/mL cerium oxide nanoenzyme which could significantly promote the proliferation of human skin fibroblasts and had high superoxide dismutase activity was screened out. It was added to GelMA hydrogel to prepare composite hydrogel. The percentage of cerium oxide nanoenzyme released from the composite hydrogel was calculated after immersing it in phosphate buffer solution (PBS) for 3 and 7 d. The red blood cell suspension of mice was divided into PBS group, Triton X-100 group, cerium oxide nanoenzyme group, GelMA hydrogel group, and composite hydrogel group, which were treated with corresponding solution. The hemolysis of red blood cells was detected by microplate reader after 1 h of treatment. The bacterial concentrations of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli were determined after being cultured with PBS, cerium oxide nanoenzyme, GelMA hydrogel, and composite hydrogel for 2 h. The sample size in all above experiments was 3. Twenty-four 8-week-old male BALB/c mice were taken, and a full-thickness skin defect wound was prepared in the symmetrical position on the back and infected with MRSA. The mice were divided into control group without any drug intervention, and cerium oxide nanoenzyme group, GelMA hydrogel group, and composite hydrogel group applied with corresponding solution, with 6 mice in each group. The wound healing was observed on 3, 7, and 14 d after injury, and the remaining wound areas on 3 and 7 d after injury were measured (the sample size was 5). The concentration of MRSA in the wound exudation of mice on 3 d after injury was measured (the sample size was 3), and the blood flow perfusion in the wound of mice on 5 d after injury was observed using a laser speckle flow imaging system (the sample size was 6). On 14 d after injury, the wound tissue of mice was collected for hematoxylin-eosin staining to observe the newly formed epithelium and for Masson staining to observe the collagen situation (the sample size was both 3). Results: After immersion for 3 and 7 d, the release percentages of cerium oxide nanoenzyme in the composite hydrogel were about 39% and 75%, respectively. After 1 h of treatment, compared with that in Triton X-100 group, the hemolysis of red blood cells in PBS group, GelMA hydrogel group, cerium oxide nanoenzyme group, and composite hydrogel group was significantly decreased (P<0.05). Compared with that cultured with PBS, the concentrations of MRSA and Escherichia coli cultured with cerium oxide nanoenzyme, GelMA hydrogel, and composite hydrogel for 2 h were significantly decreased (P<0.05). The wounds of mice in the four groups were gradually healed from 3 to 14 d after injury, and the wounds of mice in composite hydrogel group were all healed on 14 d after injury. On 3 and 7 d after injury, the remaining wound areas of mice in composite hydrogel group were (29±3) and (13±5) mm2, respectively, which were significantly smaller than (56±12) and (46±10) mm2 in control group and (51±7) and (38±8) mm2 in cerium oxide nanoenzyme group (with P values all <0.05), but was similar to (41±5) and (24±9) mm2 in GelMA hydrogel group (with P values both >0.05). On 3 d after injury, the concentration of MRSA on the wound of mice in composite hydrogel group was significantly lower than that in control group, cerium oxide nanoenzyme group, and GelMA hydrogel group, respectively (with P values all <0.05). On 5 d after injury, the volume of blood perfusion in the wound of mice in composite hydrogel group was significantly higher than that in control group, cerium oxide nanoenzyme group, and GelMA hydrogel group, respectively (P<0.05). On 14 d after injury, the wound of mice in composite hydrogel group basically completed epithelization, and the epithelization was significantly better than that in the other three groups. Compared with that in the other three groups, the content of collagen in the wound of mice in composite hydrogel group was significantly increased, and the arrangement was also more orderly. Conclusions: The composite hydrogel has good biocompatibility and antibacterial effect in vivo and in vitro. It can continuously sustained release cerium oxide nanoenzyme, improve wound blood perfusion in the early stage, and promote wound re-epithelialization and collagen synthesis, therefore promoting the healing of infected full-thickness skin defect wounds in mice.

Growing Salicornia europaea L. with Saline Hydroponic or Aquaculture Wastewater

Among halophyte plants, Salicornia species (also known as glasswort or sea asparagus) are increasingly grown in open fields and greenhouses for edible or non-edible purposes. Their salinity tolerance makes it possible to irrigate Salicornia plants with saline waters and even seawater, which cannot be used by other crop species. In this work, S. europaea (L.) was cultivated in pots under the typical climatic conditions of the fall season in the Mediterranean region and irrigated with non-saline standard nutrient solution (SNS) or saline wastewater discharged from a greenhouse semi-closed hydroponic (substrate) culture of tomato or a saltwater recirculating aquaculture system (RAS) with Gilthead sea bream (Spaurus aurata L., which was used as such or after dilution (50:50) with SNS. Plant growth was not significantly affected by the composition of irrigation water, while higher antioxidant capacity (measured using the DPPH assay) and concentration of photosynthetic pigments, phenols, flavonoids, and ascorbic acid were found in the shoots of SNS plants than in those of plants irrigated with wastewater. The level of lipid peroxidation and H2O2 production significantly increased in the SNS plants, which also showed higher activity of superoxide dismutase and lower activity of catalase. These results suggest that S. europaea can be cultivated using wastewater with moderate to high salinity discharged from greenhouse hydroponic crops or RASs, and that salt is not strictly required for the growth of this species. Using non-saline nutrient solution can result in moderate oxidative stress that improves the shoot quality of S. europaea.

Ameliorative role of diets fortified with Artocarpus altilis in a Drosophila melanogaster model of aluminum chloride-induced neurotoxicity

Backgrounds: Artocarpus altilis (breadfruit) belongs to the family Moraceae. Artocarpus altilis possesses antioxidative, anti-inflammatory, and anti-proliferative properties. Aluminum (Al) is extensively utilized for consumer products, cooking utensils, pharmaceuticals, and industries. Indication for the neurotoxicity of Al is investigated in various studies, notwithstanding the precise mechanisms of Al toxicity are yet to be fully elucidated, and, which requires novel therapy. In this study, we determined the ameliorative role of Artocarpus altilis on aluminum chloride-induced neurotoxicity in Drosophila melanogaster.MethodsVarying concentration of the extract were used to formulate diets for 6 groups of flies. Group 1 contained basal diet, group 2 contained basal diet and aluminium chloride (AlCl3), group 3 contained basal diet + 0.1% unseeded breadfruit (UBF), group 4 contained basal diet + 1% unseeded breadfruit, group 5 and 6 contained basal diet + AlCl3 + 0.1% and 1% unseeded breadfruit. Assays such as acetylcholinesterase activity, malondialdehyde (MDA) concentration level, catalase activity, and superoxide dismutase (SOD) activity were carried out after 7 days of exposure respectively.ResultsThe results showed low activity of acetylcholinesterase activity and MDA level and high catalase and SOD activity in the pretreated and post-treated flies with Artocarpus altilis compared to the normal and negative control respectively.ConclusionsTaken together, Artocarpus altilis is a promising prophylactic, antiacetylcholinesterase, and antioxidant plant in the prevention, management and treatment of neurodegenerative diseases.Graphical

Effects of Photoperiod on Survival, Growth, Physiological, and Biochemical Indices of Redclaw Crayfish (Cherax quadricarinatus) Juveniles

Through a 30-day experiment, this study investigated the effects of five photoperiods (0L:24D, 6L:18D, 12L:12D, 18L:6D, and 24L:0D) on the survival, enzyme activity, body color, and growth-related gene expression of redclaw crayfish (Cherax quadricarinatus) juveniles. The results showed that C. quadricarinatus juveniles under 18L:6D and 24L:0D photoperiods exhibited the highest survival rate, which was significantly higher than the survival rates of juveniles under the other three photoperiods (p &lt; 0.05). However, the 0L:24D group had the highest final body weight and weight gain rate, significantly surpassing those of the 12L:12D, 18L:6D, and 24L:0D groups (p &lt; 0.05). Regarding enzyme activity and hormone levels, juveniles under the 18L:6D photoperiod exhibited relatively higher activity of superoxide dismutase (SOD), acid phosphatase (ACP), and lysozyme (LZM) enzymes than those under other photoperiods, but their levels of melatonin and cortisol were relatively low. In addition, the 24L:0D group showed the highest malondialdehyde (MDA) content. Analysis of gene expression levels revealed that retinoid X receptor (RXR) and α-amylase (α-AMY) genes in C. quadricarinatus juveniles exhibited significantly higher expression levels under the 18L:6D photoperiod than those under the other four photoperiods (p &lt; 0.05). With increasing daylight exposure, the body color of C. quadricarinatus changed from pale blue to yellow–brown. In summary, C. quadricarinatus juveniles achieved high survival rates, good growth performance, strong antioxidant stress response, and immune defense capabilities under an 18 h photoperiod. Therefore, in the industrial seedling cultivation of redclaw crayfish, it is recommended to provide 18 h of daily light. Further, the study demonstrated the ability to manipulate the body color of C. quadricarinatus through controlled artificial photoperiods. These findings provide essential technical parameters needed for the industrial cultivation of C. quadricarinatus juveniles.

Effect of irrigation on wild and inbred maize with relation to the antioxidant status of pollens, flag leaves, and developing grains.

The investigation was carried out to evaluate the net effect of limited irrigation on the antioxidant status of pollens, flag leaves, and developing grains of wild and inbred maize lines. Teosinte pollens showed the highest activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione-s-transferase (GST), and peroxidase (POX) under stressful conditions while LM 11 showed a significant decrease in APX, CAT, GR, and GST activities. Limited irrigations increased the contents of superoxide and malondialdehyde (MDA) to maximum levels in LM 11 leaves. The pollens, leaves, and developing grains of teosinte had the highest content of total phenols. Proline was maximum in the developing grains of teosinte and CML 32 while lowest in those of LM 11. Principal component analysis showed that LM 11 genotype and the respective antioxidant enzymes were in completely opposite quadrants. Chord analysis showed that CAT activity and total phenol content in pollens, leaves, and developing grains contributed towards most of the variations observed in teosinte and might be responsible for managing the yield attributes of genotype during stress conditions. The pollens and leaves of teosinte, with significant SOD activity, further helped in optimizing plant yield, under stressful conditions. CML 32 occupied intermediate position owing to the unaffected activities of most of the antioxidant enzymes and high content of antioxidants in its tissues. It may be concluded that the overall antioxidant status of tissues decides the tolerance behavior of plants.

Shaping the life in karst: Antioxidative response of two Balkan endemic Scilla species

Scilla litardierei and S. lakusicii are closely related species endemic to the Dinarides. While S. litardierei is a characteristic species of wet meadows in karst fields, S. lakusicii inhabits open rocky habitats in the surroundings of karst fields. During their life cycle, plants in karst areas are subjected to the influence of drought, waterlogging, high temperature and high intensity of light. The goal of our investigation was a comparative analysis of the isoenzyme profiles and activities of superoxide dismutase (SOD) and Class III peroxidase (POX) in the roots, leaves and flowers of S. litardierei and S. lakusicii that grew in different karst localities within their native range in the Dinarides. Field research was performed on karst fields and neighboring karst areas in eastern Herzegovina (Bosnia and Herzegovina) during the vegetation season in 2017. SODs are key enzymes in plant antioxidant defense, while POXs, in addition to their antioxidant role, play an important role in plant growth and development. The highest protein concentration in roots was measured for S. litardierei from the Lastva locality (13.416 ± 4.213 mg/gFW), while the lowest was measured in S. lakusicii from Popovo Polje (2.516 ± 0.565 mg/gFW). The highest SOD activity was measured for all three plants' organs from Fatničko Polje. The highest POX activity in leaf samples was measured in S. litardierei from Dabarsko Polje (0.6012 ± 0.210 μmol/mg*min), in the flower samples also in S. litardierei from Dabarsko Polje (0.288 ± 0.005 μmol/mg*min), while in root samples the highest POX activity was measured in S. litardierei from Fatničko Polje. SOD and POX isoenzyme profiles vary depending on Scilla species, locality and plant organ. According to our results, SOD and POX could play an important role in the adaptive mechanisms of Scilla species in their natural karst habitats.

Metabolic Mechanisms Underlying Heat and Drought Tolerance in Lentil Accessions: Implications for Stress Tolerance Breeding

Climate change has significantly exacerbated the effects of abiotic stresses, particularly high temperatures and drought stresses. This study aims to uncover the mechanisms underlying heat and drought tolerance in lentil accessions. To achieve this objective, twelve accessions were subjected to high-temperature stress (32/20 °C), while seven accessions underwent assessment under drought stress conditions (50% of field capacity) during the reproductive stage. Our findings revealed a significant increase in catalase activity across all accessions under both stress conditions, with ILL7814 and ILL7835 recording the highest accumulations of 10.18 and 9.33 under drought stress, respectively, and 14 µmol H2O2 mg protein−1 min−1 under high temperature. Similarly, ascorbate peroxidase significantly increased in all tolerant accessions due to high temperatures, with ILL6359, ILL7835, and ILL8029 accumulating the highest values with up 50 µmol ascorbate mg protein−1 min−1. In contrast, no significant increase was obtained for all accessions subjected to water stress, although the drought-tolerant accessions accumulated more APX activity (16.59 t to 25.08 µmol ascorbate mg protein−1 min−1) than the sensitive accessions. The accessions ILL6075, ILL7814, and ILL8029 significantly had the highest superoxide dismutase activity under high temperature, while ILL6363, ILL7814, and ILL7835 accumulated the highest values under drought stress, each with 22 to 25 units mg protein−1. Under both stress conditions, ILL7814 and ILL7835 recorded the highest contents in proline (38 to 45 µmol proline/g FW), total flavonoids (0.22 to 0.77 mg QE g−1 FW), total phenolics (7.50 to 8.79 mg GAE g−1 FW), total tannins (5.07 to 20 µg CE g−1 FW), and total antioxidant activity (60 to 70%). Further, ILL7814 and ILL6338 significantly recorded the highest total soluble sugar content under high temperature (71.57 and 74.24 mg g−1, respectively), while ILL7835 achieved the maximum concentration (125 mg g−1) under drought stress. The accessions ILL8029, ILL6104, and ILL7814 had the highest values of reducing sugar under high temperature with 0.62 to 0.79 mg g−1, whereas ILL6075, ILL6363, and ILL6362 accumulated the highest levels of this component under drought stress with 0.54 to 0.66 mg g−1. Overall, our findings contribute to a deeper understanding of the metabolomic responses of lentil to drought and heat stresses, serving as a valuable reference for lentil stress tolerance breeding.

Genome-wide identification of the LRR-RLK gene family in peanut and functional characterization of AhLRR-RLK265 in salt and drought stresses

Leucine-rich repeat receptor-like kinases (LRR-RLKs) play important roles in plant developmental regulations and various stress responses. Peanut (Arachis hypogaea L.) is a worldwide important oil crop; however, no systematic identification or analysis of the peanut LRR-RLK gene family has been reported. In present study, 495 LRR-RLK genes in peanut were identified and analyzed. The 495 AhLRR-RLK genes were classed into 14 groups and 10 subgroups together with their Arabidopsis homologs according to phylogenetic analyses, and 491 of 495 AhLRR-RLK genes unequally located on 20 chromosomes. Analyses of gene structure and protein motif organization revealed similarity in exon/intron and motif organization among members of the same subgroup, further supporting the phylogenetic results. Gene duplication events were found in peanut LRR-RLK gene family via syntenic analysis, which were important in LRR-RLK gene family expansion in peanut. We found that the expression of AhLRR-RLK genes was detected in different tissues using RNA-seq data, implying that AhLRR-RLK genes may differ in function. In addition, Arabidopsis plants overexpressing stress-induced AhLRR-RLK265 displayed lower seed germination rates and root lengths compared to wild-type under exogenous ABA treatment. Notably, overexpression of AhLRR-RLK265 enhanced tolerance to salt and drought stresses in transgenic Arabidopsis. Moreover, the AhLRR-RLK265-OE lines were found to have higher activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) under salt and drought stress treatments. We believe these results may provide valuable information about the function of peanut LRR-RLK genes for further analysis.

Physiological and antioxidant responses associated with Sub1 gene introgressed rice (Oryza sativa L.) lines under complete submergence.

Rice is the only crop which is well adapted to aquatic environment but, it is unable to survive if completely submerged for several weeks. Breeding rice varieties with submergence tolerance is one of the best approaches to alleviate the adverse effect of submergence which requires the introgression of Sub1 gene into elite rice varieties. Hence, the study was undertaken to introgress submergence tolerant gene into the rice variety Jaya through Marker-Assisted Backcross Breeding. Also the physiological and biochemical responses like survival percentage, underwater shoot elongation, total carbohydrate content and superoxide dismutase activity were also studied in Sub1 introgressed lines. We could develop twenty Sub1 introgressed lines with Sub1 region of 3.1-5.1mb and with 80.0- 95.3% recurrent parent genome recovery. Sub1 introgressed Jaya lines and the tolerant checks FR13A and Swarna Sub1 had lower shoot elongation under water, higher superoxide dismutase activity (about 5 times) upto 4h after de-submergence which resulted in higher survival percentage. The reduced shoot elongation of tolerant varieties reduced the utilization of stored carbohydrate. Through our research we introgressed Sub1 gene into rice variety Jaya through Marker-Assisted Backcross Breeding and could study the physiological responses under submergence by which we confirmed the presence of Sub1 gene in these lines. These lines could be field evaluated and could be released as a new variety thus helping the farmers of flood prone areas of Kerala.

Dietary calcium requirement of bullfrog (Lithobates catesbeiana)

This study was conducted to determine dietary calcium (Ca) requirement of bullfrog (Lithobates catesbeiana). Six diets were formulated to contain 0.55% (control), 0.96%, 1.30%, 1.72%, 2.02% and 2.53% Ca. Each diet was fed to triplicate tanks of bullfrogs (24.96 ± 0.03 g), and each tanks was stocked with 13 bullfrogs. Bullfrogs were fed twice daily (8:30 and 17:30) to apparent satiation for 8 weeks. Weight gain rate of bullfrogs fed the 1.72% Ca diet was significantly higher than the control group (P < 0.05). The content of Ca was significantly increased in the whole body and vertebrae of bullfrogs fed the 1.72%, 2.02% and 2.53% diets compared to those fed the 0.55%, 0.96% and 1.30% Ca diets (P < 0.05). Furthermore, bullfrogs fed the 1.30%, 1.72% and 2.02% diets had higher activity of trypsin than those fed the 0.55% and 2.53% Ca diets (P < 0.05). Apparent digestibility coefficient of dry matter was higher in bullfrogs fed the 1.72% Ca diet than those fed the 0.55% and 2.53% diets (P < 0.05). Bullfrogs fed the 1.72% Ca diet had higher activity of superoxide dismutase in the serum than those fed the 0.55% and 2.53% diets (P < 0.05). Compared to the control group, contents of 1,25 dihydroxyvitamin D3 and parathyroid hormone in the serum as well as vitamin D receptor and sodium-calcium exchanger genes expression in the kidney were significantly decreased in bullfrogs fed the 1.72%, 2.02% and 2.53% Ca diets (P < 0.05). Compared to the control group, intestinal sodium-calcium exchanger gene expression was significantly decreased in bullfrogs fed the 1.72% and 2.02% Ca diets (P < 0.05). Based on second order polynomial analysis of weight gain rate, dietary Ca requirement of bullfrog was 1.89%.

Sufentanil reduces myocardial apoptosis in rats with myocardial ischemia-reperfusion injury

Purpose: To determine the effect of sufentanil on myocardial apoptosis in rats with myocardial ischemia-reperfusion injury (MIRI).Methods: Fifty Sprague Dawley rats were randomly assigned to five groups: sham, model, low-dose, moderate-dose, and high-dose. The groups, except sham, underwent ligation of the left anterior descending coronary artery to establish the MIRI model. The low, moderate, and high-dose groups received intraperitoneal injections of sufentanil at different concentrations. Cardiac function, serum LDH, creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) content were evaluated. The mRNA expression levels of apoptosis genes and protein levels of p38 and p-p38 were assessed in myocardial tissues using various methods while apoptosis was assessed by TUNEL assay.Results: Compared to sham group, the model group exhibited significant decrease in fractional shortening (FS) and ejection fraction (EF), increase in CK activity, LDH, and MDA contents, lower SOD activity. Model group also showed increase in mRNA levels of B-cell lymphoma-2 (Bcl-2) and caspase- 3, higher apoptosis, significant increase in protein levels of p38 and p-p38, and higher level of myocardial apoptosis (p &lt; 0.05). High-dose group demonstrated significant increase in FS and EF, decrease in LDH content and CK activity, lower MDA content, higher SOD activity, decrease in mRNA levels of Bcl-2 and caspase-3, lower apoptosis, decrease in protein levels of p38 and p-p38, and lower level of myocardial apoptosis (p &lt; 0.05), when compared with model group.Conclusion: High-dose sufentanil reduces myocardial apoptosis and improves cardiac function, and thus can potentially be developed as a cardioprotective agent.

Winter and Summer Variations in the Physiological Parameters of Two Scleractinian Corals in Sanya Bay

Coral reefs in Sanya Bay have been degrading in recent decades under climate change and human activities. To identify physiological changes of scleractinian corals and corresponding influencing factors, aquatic environmental factors and physiological parameters of Pocillopora damicornis, Porites pukoensis and their symbiotic zooxanthellae were examined in four Sanya Bay coral reef areas in December 2020 (winter) and July 2021 (summer). The density and chlorophyll a+c2 content of the symbiotic zooxanthellae were significantly high in winter and low in summer. Superoxide dismutase and caspase3 activities of corals and zooxanthellae were high in summer and low in winter, whereas catalase activity showed the opposite pattern. The variations were consistent for both coral symbionts. Water temperature and salinity were the main factors affecting the physiological variations of corals. Compared with winter, the high temperature/low salinity aquatic environment in summer reduced the density and chlorophyll a+c2 content of zooxanthellae, resulting in high superoxide dismutase and caspase3 activities in the corals and zooxanthellae. In addition, turbidity was an important factor affecting the physiological characteristics of coral–zooxanthellae symbionts among the four coral reef areas. Our results have important implications for understanding the changes in coral reef communities in Sanya Bay and coral reef protection.

Enhancement of superoxide dismutase activity using mixed Lactobacillus casei and Saccharomyces cerevisiae cultures in simulated gastrointestinal conditions with encapsulation

Superoxide dismutase (SOD)-producing microorganisms have attracted increasing attention given their scavenge superoxide anions capabilities, but further strengthening methods are still urgently needed. Herein, Lactobacillus casei F18 and Saccharomyces cerevisiae J14 were isolated for their high SOD activity. Co-culturing these two strains emerged effective nutritional cooperation, which increased SOD activity by 198.22% and 66.35% compared to F18 and J14, respectively. SOD activity of co-culture “F18 +J14” was further enhanced by response surface methodology and metal ion stress. High-performance liquid chromatographic and RT-qPCR analyses of “F18 +J14” revealed that SOD production pathways were active and contents of metabolites acetate and ethanol were dramatically increased. Metabolite complementarity experiment also verified the addition of acetate and ethanol in vitro enhanced SOD activity and gene expression of “F18 +J14”. These findings highlighted co-culture and metal ion stress enhanced SOD activity by cross-stimulating cells with metabolites, affecting cell growth, and up-regulating SOD genes. Furthermore, “F18 +J14” based immobilized bio-beads were successfully synthesized with sodium alginate combined CaCO3, which exhibited the highest cell vitality and SOD activity in the simulated gastrointestinal conditions. Together, this study revealed the synergistic promotion mechanism of SOD biosynthesis by co-cultivation lactic acid bacteria and yeast, and shows great promise for management of oxidation stress related diseases.

PbrWRKY70 increases pear (Pyrus bretschneideri Rehd) black spot disease tolerance by negatively regulating ethylene synthesis via PbrERF1B-2

Various pear plant cultivars exhibit diverse abilities to resist pear black spot disease (BSD), while the precise molecular mechanisms of resistance against pear BSD remain unclear. This study proposed a profound expression of a WRKY gene, namely PbrWRKY70, derived from Pyrus bretschneideri Rehd, within a BSD-resistant pear cultivar. Comparative analysis against the wild-type revealed that the overexpression of PbrWRKY70 engendered augmented BSD resistance of transgenic Arabidopsis thaliana and pear calli. Notably, the transgenic plants exhibited higher activities of superoxide dismutase and peroxidase, along with an elevated capacity to counteract superoxide anions via increased anti-O2-. Additionally, these plants displayed diminished lesion diameter, as well as reduced levels of hydrogen peroxide, malondialdehyde and 1-aminocyclopropane-1-carboxylic acid (ACC) contents. We subsequently demonstrated that PbrWRKY70 selectively bound to the promoter region of ethylene-responsive transcription factor 1B-2 (PbrERF1B-2), a potential negative regulator of ACC, thereby downregulating the expression of ACC synthase gene (PbrACS3). Consequently, we confirmed that PbrWRKY70 could enhance pear resistance against BSD by reducing ethylene production via modulation of the PbrERF1B-2-PbrACS3 pathway. This study established the pivotal relationship among PbrWRKY70, ethylene synthesis and pear BSD resistance, fostering the development of novel BSD-resistant cultivars. Furthermore, this breakthrough holds the potential to enhance pear fruit yield and optimize storage and processing during the later stages of fruit maturation.