Ascorbate Peroxidase Gene Research Articles

A high-quality genome of Actinidia eriantha provides new insight into ascorbic acid regulation

Actinidia eriantha is one of the species of kiwifruit with a particularly high ascorbic acid (AsA) content. However, the molecular mechanism driving AsA richness in fruit remains unclear. In order to reveal the molecular mechanism of AsA richness in A. eriantha, this study constructed a regulatory network related to AsA metabolism by combining genomics, metabolomics and transcriptomics. We assembled a high-quality genome of A. eriantha ‘Ganlv 1’ with only five remaining gaps. The assembly is comprised of 29 pseudochromosomes with a total size of 615.95 Mb, and contig N50 of 20.35 Mb. Among them, 24 of the pseudochromosomes were obtained directly from telomere-to-telomere. The LTR assembly index score and consensus quality value were 21.34 and 39.90%, respectively. Subsequently, 61 metabolites and 2 092 genes were found to be differentially accumulated/expressed during fruit development by metabolome and transcriptome assays, respectively. AsA metabolism and the cyclic regeneration pathway were found to have high expression levels throughout fruit growth and development, suggesting its crucial role in the regulation of AsA. Furthermore, the AsA contents are highly associated with ascorbate peroxidase genes. The genome obtained in this study provides genomic resources for the genetic and breeding research of A. eriantha, and the constructed regulatory network can provide a public data platform for future research on kiwifruit.

Genome-wide identification and expression profiling of APX gene family under multifactorial stress combinations and melatonin-mediated tolerance in pitaya

Pitaya (Selenicereus undatus L.), an important tropical plant belonging to the Cactaceae family, is rich in antioxidants, dietary fibers and low in calories. Pitaya shows higher tolerance against abiotic stresses; however, various environmental factors lead to excessive production of reactive oxygen species (ROS) persistently that cause injury to the cells. Under vanadium, drought, and heat conditions, plant cells regulate the expression of ascorbate peroxidase (APX, a vital antioxidant enzyme) genes, which play a significant role in detoxifying ROS by catalyzing hydrogen peroxide. In this study, we have identified 65 APX genes (HuAPX) in the pitaya genome and clustered them into 13 subfamilies. Gene structure and motif analysis revealed that HuAPX genes contained well-preserved intron-exon structure and their motif domains also shared the same order within one cluster. Phytohormones-related cis-acting elements including abscisic acid, gibberellin, jasmonic acid, salicylic acid, and auxin were found in abundance in the promoter regions of the HuAPX genes. RNA-seq-based expression analysis of the HuAPX genes unveiled that few genes showed significantly higher expression under single and multifactorial stress combinations of drought, heat, and vanadium. Exogenous applications of melatonin attenuate the vanadium, heat, and drought effects, thus enhancing the growth and development of the pitaya plant, by inducing the HuAPX gene expression efficiently. Moreover, the expression pattern of HuAPX genes were validated through qRT-PCR analysis. Our findings suggest that three candidate genes (HuAPX1/3/6) may play a key role against abiotic stresses by encoding key enzymes related to glutathione metabolism and phenylpropanoid biosynthesis. These results provide valuable information for organizing a new strategy to design the climate-smart pitaya genotypes through conventional and modern biotechnological tools.

Ascorbate peroxidase plays an important role in photoacclimation in the extremophilic red alga Cyanidiococcus yangmingshanensis.

Acidothermophilic cyanidiophytes in natural habitats can survive under a wide variety of light regimes, and the exploration and elucidation of their long-term photoacclimation mechanisms promises great potential for further biotechnological applications. Ascorbic acid was previously identified as an important protectant against high light stress in Galdieria partita under mixotrophic conditions, yet whether ascorbic acid and its related enzymatic reactive oxygen species (ROS) scavenging system was crucial in photoacclimation for photoautotrophic cyanidiophytes was unclear. The significance of ascorbic acid and related ROS scavenging and antioxidant regenerating enzymes in photoacclimation in the extremophilic red alga Cyanidiococcus yangmingshanensis was investigated by measuring the cellular content of ascorbic acid and the activities of ascorbate-related enzymes. Accumulation of ascorbic acid and activation of the ascorbate-related enzymatic ROS scavenging system characterized the photoacclimation response after cells were transferred from a low light condition at 20 μmol photons m-2 s-1 to various light conditions in the range from 0 to 1000 μmol photons m-2 s-1. The activity of ascorbate peroxidase (APX) was most remarkably enhanced with increasing light intensities and illumination periods among the enzymatic activities being measured. Light-dependent regulation of the APX activity was associated with transcriptional regulation of the chloroplast-targeted APX gene. The important role of the APX activity in photoacclimation was evidenced by the effect of the APX inhibitors on the photosystem II activity and the chlorophyll a content under the high light condition at 1000 μmol photons m-2 s-1. Our findings provide a mechanistic explanation for the acclimation of C. yangmingshanensis to a wide range of light regimes in natural habitats.

Genome-wide characterization of ascorbate peroxidase gene family in pepper (Capsicum annuum L.) in response to multiple abiotic stresses.

Pepper is widely grown all over the world, so it faces many abiotic stresses, such as drought, high temperature, low temperature, salt damage, and so on. Stresses causing the accumulation of reactive oxidative species (ROS) in plants are removed by antioxidant defense systems, and ascorbate peroxidase (APX) is an important antioxidant enzyme. Therefore, the present study performed genome-wide identification of the APX gene family in pepper. We identified nine members of the APX gene family in the pepper genome according to the APX proteins' conserved domain in Arabidopsis thaliana. The physicochemical property analysis showed that CaAPX3 had the longest protein sequence and the largest molecular weight of all genes, while CaAPX9 had the shortest protein sequence and the smallest MW. The gene structure analysis showed that CaAPXs were composed of seven to 10 introns. The CaAPX genes were divided into four groups. The APX genes of groups I and IV were localized in the peroxisomes and chloroplasts, respectively; the group II genes were localized in the chloroplasts and mitochondria; and the group III genes were located in the cytoplasm and extracell. The conservative motif analysis showed that all APX genes in the pepper had motif 2, motif 3, and motif 5. The APX gene family members were distributed on five chromosomes (Chr. 2, 4, 6, 8, and 9). The cis-acting element analysis showed that most CaAPX genes contain a variety of cis-elements related to plant hormones and abiotic stress. RNA-seq expression analysis showed that the expression patterns of nine APXs were different in vegetative and reproductive organs at different growth and development stages. In addition, the qRT-PCR analysis of the CaAPX genes revealed significant differential expression in response to high temperature, low temperature, and salinity stresses in leaf tissue. In conclusion, our study identified the APX gene family members in the pepper and predicted the functions of this gene family, which would provide resources for further functional characterization of CaAPX genes.

Stress memory in seedlings of primed seed chickpea (Cicer arietinum L.) using sodium nitroprusside under cold stress

Pea cultivation has been developed as a rich source of protein in many countries. The spring cultivation of chickpeas has low performance and efficiency due to the short length of the growth period and the confrontation of the reproductive period with drought and high temperature stress. Autumn cultivation of chickpeas is introduced as an important solution in reducing the yield caused by high temperature and drought stress during the flowering time of spring cultivation. However, in autumn cultivation, sensitivity to cold is the main limitation. So, the present study was performed to investigate the effect of nitric oxide on stress memory due to seed priming with sodium nitroprusside for cold tolerance of chickpea seedlings under simulated autumn temperatures. This experiment was conducted as a factorial in a completely randomized design with nine replicates at the Research Center for Plant Sciences of Ferdowsi University, Mashhad Iran, in 2020. Experimental factors included seed priming at two levels (control and sodium nitroprusside) and chickpea cultivars, including a susceptible genotype (MCC505) and a cold-tolerant cultivar (Saral). This experiment evaluated the gene expression of antioxidant enzymes (superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase) in the seedling stage. The results revealed that superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase genes were expressed in sodium nitroprusside and control treatments. In the Saral cultivar, sodium nitroprusside increased the gene expression of superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase enzymes by 29, 88, 54, and 63%, respectively. On the other hand, the gene expression of these enzymes in the control treatment may be due to the effect of temperature on them. There was a significant increase in the expression of superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase genes in the Saral cultivar compared to the MCC505 genotype. In general, sodium nitroprusside increased the activity of antioxidant enzymes in the Saral cultivar through the expression of antioxidant enzyme genes.

Conserved hierarchical gene regulatory networks for drought and cold stress response in Myrica rubra.

Stress response in plant is regulated by a large number of genes co-operating in diverse networks that serve multiple adaptive process. To understand how gene regulatory networks (GRNs) modulating abiotic stress responses, we compare the GRNs underlying drought and cold stresses using samples collected at 4 or 6h intervals within 48h in Chinese bayberry (Myrica rubra). We detected 7,583 and 8,840 differentially expressed genes (DEGs) under drought and cold stress respectively, which might be responsive to environmental stresses. Drought- and cold-responsive GRNs, which have been built according to the timing of transcription under both abiotic stresses, have a conserved trans-regulator and a common regulatory network. In both GRNs, basic helix-loop-helix family transcription factor (bHLH) serve as central nodes. MrbHLHp10 transcripts exhibited continuous increase in the two abiotic stresses and acts upstream regulator of ASCORBATE PEROXIDASE (APX) gene. To examine the potential biological functions of MrbHLH10, we generated a transgenic Arabidopsis plant that constitutively overexpresses the MrbHLH10 gene. Compared to wild-type (WT) plants, overexpressing transgenic Arabidopsis plants maintained higher APX activity and biomass accumulation under drought and cold stress. Consistently, RNAi plants had elevated susceptibility to both stresses. Taken together, these results suggested that MrbHLH10 mitigates abiotic stresses through the modulation of ROS scavenging.

High-Nitrate-Supply-Induced Transcriptional Upregulation of Ascorbic Acid Biosynthetic and Recycling Pathways in Cucumber

Nowadays open field and protected vegetable cultivation practices require and use genotypes which are precisely tailored to their intended growth environments. Variability of this kind provides a rich source of material to uncover molecular mechanisms supporting the necessarily divergent physiological traits. In this study, typical field-optimized and glasshouse-cultivated cucumber F1 hybrids were investigated, and displayed slower growth ('Joker') and faster growth ('Oitol') in seedlings. Antioxidant capacity was lower in 'Joker' and higher in 'Oitol', pointing to a potential redox regulation of growth. The growth response of seedlings to paraquat treatment indicated stronger oxidative stress tolerance in the fast-growing 'Oitol'. To test whether protection against nitrate-induced oxidative stress was also different, fertigation with increasing potassium nitrate content was applied. This treatment did not change growth but decreased the antioxidant capacities of both hybrids. Bioluminescence emission revealed stronger lipid peroxidation triggered by high nitrate fertigation in the leaves of 'Joker' seedlings. To explore the background of the more effective antioxidant protection of 'Oitol', levels of ascorbic acid (AsA), as well as transcriptional regulation of relevant genes of the Smirnoff-Wheeler biosynthetic pathway and ascorbate recycling, were investigated. Genes related to AsA biosynthesis were strongly upregulated at an elevated nitrate supply in 'Oitol' leaves only, but this was only reflected in a small increase in total AsA content. High nitrate provision also triggered expression of ascorbate-glutathion cycle genes with stronger or exclusive induction in 'Oitol'. AsA/dehydro-ascorbate ratios were higher in 'Oitol' for all treatments, with a more pronounced difference at high nitrate levels. Despite strong transcriptional upregulation of ascorbate peroxidase genes (APX) in 'Oitol', APX activity only increased significantly in 'Joker'. This suggests potential inhibition of APX enzyme activity specifically in 'Oitol' at a high nitrate supply. Our results uncover an unexpected variability in redox stress management in cucumbers, including nitrate inducibility of AsA biosynthetic and recycling pathways in certain genotypes. Possible connections between AsA biosynthesis, recycling and nitro-oxidative stress protection are discussed. Cucumber hybrids emerge as an excellent model system for studying the regulation of AsA metabolism and the roles of AsA in growth and stress tolerance.

Identification of citrus APX gene family and their response to CYVCV infection.

Ascorbate peroxidase (APX) is one of the most important antioxidant enzymes in the reactive oxygen metabolic pathway of plants. The role of APX under biotic and abiotic stress conditions has been explored, but the response pattern of APX under biotic stresses is relatively less known. In this study, seven CsAPXs gene family members were identified based on the sweet orange (Citrus sinensis) genome and subjected to evolutionary and structural analysis using bioinformatics software. The APX genes of lemon (ClAPXs) were cloned and showed a high conservation to CsAPXs by sequences alignment. In citrus yellow vein clearing virus (CYVCV)-infected Eureka lemons (C. limon) at 30th day post inoculation, APX activity and accumulation of hydrogen peroxide (H2O2) and malondialdehyde were measured to be 3.63, 2.29, and 1.73 times to that of the healthy control. The expression levels of 7 ClAPX genes in different periods of CYVCV-infected Eureka lemon were analyzed. Notably, ClAPX1, ClAPX5, and ClAPX7 showed higher expression levels compared to healthy plants, while ClAPX2, ClAPX3, and ClAPX4 showed lower expression levels. Functional identification of ClAPX1 in Nicotiana benthamiana showed that increasing the expression of ClAPX1 could significantly reduce the accumulation of H2O2, and it was verified that ClAPX1 is located in the plasma membrane of the cell. The present study provided information on the evolution and function of citrus APXs and revealed for the first time their response pattern to CYVCV infection.

Suppression of the HOS1 Gene Affects the Level of ROS Depending on Light and Cold.

The E3 ubiquitin-protein ligase HOS1 is an important integrator of temperature information and developmental processes. HOS1 is a negative regulator of plant cold tolerance, and silencing HOS1 leads to increased cold tolerance. In the present work, we studied ROS levels in hos1Cas9Arabidopsis thaliana plants, in which the HOS1 gene was silenced by disruption of the open reading frame via CRISPR/Cas9 technology. Confocal imaging of intracellular reactive oxygen species (ROS) showed that the hos1 mutation moderately increased levels of ROS under both low and high light (HL) conditions, but wild-type (WT) and hos1Cas9 plants exhibited similar ROS levels in the dark. Visualization of single cells did not reveal differences in the intracellular distribution of ROS between WT and hos1Cas9 plants. The hos1Cas9 plants contained a high basal level of ascorbic acid, maintained a normal balance between reduced and oxidized glutathione (GSH and GSSG), and generated a strong antioxidant defense response against paraquat under HL conditions. Under cold exposure, the hos1 mutation decreased the ROS level and substantially increased the expression of the ascorbate peroxidase genes Apx1 and Apx2. When plants were pre-exposed to cold and further exposed to HL, the expression of the NADPH oxidase genes RbohD and RbohF was increased in the hos1Cas9 plants but not in WT plants. hos1-mediated changes in the level of ROS are cold-dependent and cold-independent, which implies different levels of regulation. Our data indicate that HOS1 is required to maintain ROS homeostasis not only under cold conditions, but also under conditions of both low and high light intensity. It is likely that HOS1 prevents the overinduction of defense mechanisms to balance growth.

Antioxidant enzymes activity and gene expression in wheat-stripe rust interaction at seedling stage

The current study was designed to determine the activity of antioxidant enzymes: superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione peroxidase GPX and catalase (CAT) and the expression of their genes in 10 wheat isogenic differential lines showing compatible and incompatible interactions with Puccinia striiformis. The enzyme activity was determined at 24 h after inoculation (24hai), 48 h after inoculation (48hai), and 72 h after inoculation (72hai). However, enzyme gene expression was determined only at 72hai. To fulfill the objectives, 12-day-old wheat seedlings of isogenic differential lines were inoculated with stripe rust race (race 574232). The results of the study showed that enzyme activity and gene expression was significantly high in incompatible interaction as compared to compatible interaction. The enzyme activity of SOD was high at 24hai in resistant isogenic differential lines. High enzyme activity (97.54 ± 4.84) was recorded in was recorded Yr5/6* Avocet S. Enzyme activities of CAT and APX were higher at 72hai in resistant isogenic differential lines. CAT, APX genes were significantly upregulated and while gene expression of SOD was low at 72hai.The results indicate a unique pattern of enzyme activity and gene expression of the antioxidant enzymes in the compatible and incompatible interaction is playing a crucial role in the plant defense system.

Ascorbate-Glutathione Cycle Genes Families in Euphorbiaceae: Characterization and Evolutionary Analysis.

Ascorbate peroxidase (APX), Monodehydroascorbate Reductase (MDAR), Dehydroascorbate Reductase (DHAR) and Glutathione Reductase (GR) enzymes participate in the ascorbate-glutathione cycle, which exerts a central role in the antioxidant metabolism in plants. Despite the importance of this antioxidant system in different signal transduction networks related to development and response to environmental stresses, the pathway has not yet been comprehensively characterized in many crop plants. Among different eudicotyledons, the Euphorbiaceae family is particularly diverse with some species highly tolerant to drought. Here the APX, MDAR, DHAR, and GR genes in Ricinus communis, Jatropha curcas, Manihot esculenta, and Hevea brasiliensis were identified and characterized. The comprehensive phylogenetic and genomic analyses allowed the classification of the genes into different classes, equivalent to cytosolic, peroxisomal, chloroplastic, and mitochondrial enzymes, and revealed the duplication events that contribute to the expansion of these families within plant genomes. Due to the high drought stress tolerance of Ricinus communis, the expression patterns of ascorbate-glutathione cycle genes in response to drought were also analyzed in leaves and roots, indicating a differential expression during the stress. Altogether, these data contributed to the characterization of the expression pattern and evolutionary analysis of these genes, filling the gap in the proposed functions of core components of the antioxidant mechanism during stress response in an economically relevant group of plants.

Isolation of ascorbate peroxidase (APX) gene in lentil (Lens culinaris Medik.) and expression analysis under drought stress conditions

Objective: The objective of this study was to isolate partial cDNA that belongs to the ascorbate peroxidase (APX) gene of lentil (Lens culinaris Medik.) and to express LcAPX gene in lentil seedlings under drought stress conditions. Material and Methods: To identify the relationships between drought stress and LcAPX gene expression, lentil seedlings grown for 2 weeks were subjected to drought stress through not irrigating for 6, 13, and 20 days. Effects of drought stress were determined by measuring the stem relative water content (RWC). Gene expression changes in lentil seedlings were determined with real-time RT-qPCR. Results: The LcAPX gene expression levels of both drought-tolerant Firat-87 and drought-sensitive Ozbek cultivars varied with the severity of drought stress. The gene expression of LcAPX reached the highest level in Firat-87 cultivar on the 6th day, whereas a significant increase was observed only on the 20th day of the Ozbek cultivar, and this increase was relatively low as compared to the Fırat-87 cultivar. Conclusion: From the study conducted, it was concluded that time-dependent changes of the expression of LcAPX gene indicates that LcAPX gene had a highly specific gene expression profile and complex regulation in lentil drought response.

Effects and Molecular Mechanism of Mycorrhiza on the Growth, Nutrient Absorption, Quality of Fresh Leaves, and Antioxidant System of Tea Seedlings Suffering from Salt Stress

We studied changes in plant growth, nutrient absorption, quality of fresh leaves, and the antioxidant enzyme systems of leaves treated with AMF (Glomus etunicatum) in tea (tea cultivar “Xinyang 10”) seedlings exposed to 0 and 100 mmol/L sodium chloride (NaCl). The AMF colonization in the tea roots decreased observably by 50.1% after a 5-week soil NaCl (100 mmol/L) treatment. The growth, leaf nutrient levels, and leaf quality parameter contents significantly declined by 18–39% in the 100 mmol/L NaCl treatments. In contrast, these variables exhibited observably higher responses in the mycorrhizal seedlings than in the nonmycorrhizal seedlings. Furthermore, AMF improved the leaves’ total amino acid concentrations dramatically, accompanied by the upregulation of the genes of the amino acid synthetic enzymes, such as glutamate dehydrogenase (CsGDH), glutamate synthase (CsGOGAT), and glutamine synthetase (CsGS), while 100 mmol/L NaCl seedlings represented a negative performance. In addition, the 100 mmol/L NaCl treatments dramatically downregulated the expression level of the tea caffeine synthase 1 gene (CsTCS1), the ascorbate peroxidase gene (CsAPX), and the 3-hydroxy-3-methylglutaryl coenzyme gene (CsHMGR) in the leaves, while the AMF seedlings represented positive performances. These results suggest that AMF may play an active role in fresh leaf quality via the partial upregulation of the relevant genes’ expression. In contrast, salt stress represented the opposite result in tea. The seedlings inoculated with AMF showed significantly increased antioxidant enzyme activities, by 13.3–19.6%, including peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD), over the non-AMF inoculated tea seedlings. Still, they did not affect glutathione reductase (GR), irrespective of the NaCl condition. Further studies indicated that AMF observably upregulated the genes’ expressions (i.e., CsCAT and CsSOD) in both the 0 and 100 mmol/L NaCl seedlings. Meanwhile, the 100 mmol/L NaCl seedlings represented markedly lower antioxidant enzyme activities (i.e., SOD, CAT, and POD) and gene expressions (i.e., CsSOD and CsCAT) than the non-NaCl seedlings, irrespective of AMF inoculation. These results imply that AMF has a positive role in strengthening salt tolerance and on the quality of fresh tea leaves.

Genome-Wide Characterization of Ascorbate Peroxidase Gene Family in Peanut (Arachis hypogea L.) Revealed Their Crucial Role in Growth and Multiple Stress Tolerance.

Ascorbate peroxidase (APX), an important antioxidant enzyme, plays a significant role in ROS scavenging by catalyzing the decrease of hydrogen peroxide under various environmental stresses. Nevertheless, information about the APX gene family and their evolutionary and functional attributes in peanut (Arachis hypogea L.) was not reported. Therefore, a comprehensive genome-wide study was performed to discover the APX genes in cultivated peanut genome. This study identified 166 AhAPX genes in the peanut genome, classified into 11 main groups. The gene duplication analysis showed that AhAPX genes had experienced segmental duplications and purifying selection pressure. Gene structure and motif investigation indicated that most of the AhAPX genes exhibited a comparatively well-preserved exon-intron pattern and motif configuration contained by the identical group. We discovered five phytohormones-, six abiotic stress-, and five growth and development-related cis-elements in the promoter regions of AhAPX. Fourteen putative ah-miRNAs from 12 families were identified, targeting 33 AhAPX genes. Furthermore, we identified 3,257 transcription factors from 38 families (including AP2, ARF, B3, bHLH, bZIP, ERF, MYB, NAC, WRKY, etc.) in 162 AhAPX genes. Gene ontology and KEGG enrichment analysis confirm the role of AhAPX genes in oxidoreductase activity, catalytic activity, cell junction, cellular response to stimulus and detoxification, biosynthesis of metabolites, and phenylpropanoid metabolism. Based on transcriptome datasets, some genes such as AhAPX4/7/17/77/82/86/130/133 and AhAPX160 showed significantly higher expression in diverse tissues/organs, i.e., flower, leaf, stem, roots, peg, testa, and cotyledon. Likewise, only a few genes, including AhAPX4/17/19/55/59/82/101/102/137 and AhAPX140, were significantly upregulated under abiotic (drought and cold), and phytohormones (ethylene, abscisic acid, paclobutrazol, brassinolide, and salicylic acid) treatments. qRT-PCR-based expression profiling presented the parallel expression trends as generated from transcriptome datasets. Our discoveries gave new visions into the evolution of APX genes and provided a base for further functional examinations of the AhAPX genes in peanut breeding programs.

Caffeic acid regulated respiration, ethylene and reactive oxygen species metabolism to suppress senescence of Malus domestica

The current study assessed the variations of reactive oxygen species (ROS), respiration and ethylene metabolism in ‘Qiujin’ apples after caffeic acid dipping. Results demonstrated that caffeic acid remarkably inhibited respiratory rate, ethylene release rate and hydrogen peroxide accumulation, but increased reduced glutathione and ascorbic acid contents in apples. Caffeic acid also improved catalase, peroxidase, superoxide dismutase, nicotinamide adenine dinucleotide phosphate oxidase, dehydroascorbate reductase, monodehydroascorbate reductase, glutathione reductase and ascorbate peroxidase activities and gene expressions in apples. In addition, caffeic acid up-regulated MdG6PDH6 and MdAOX expressions, down-regulated MdACS1, MdACO2, MdPK, MdHK3, MdPFK, MdSDH and MdCCO expressions in the fruit. Taken together, caffeic acid could regulate the key enzymes activities and gene expressions in respiration, ethylene and ROS metabolism to suppress senescence of apples after harvest.

Whole-Genome Identification of APX and CAT Gene Families in Cultivated and Wild Soybeans and Their Regulatory Function in Plant Development and Stress Response.

Plants coevolved with their antioxidant defense systems, which detoxify and adjust levels of reactive oxygen species (ROS) under multiple plant stresses. We performed whole-genome identification of ascorbate peroxidase (APX) and catalase (CAT) families in cultivated and wild soybeans. In cultivated and wild soybean genomes, we identified 11 and 10 APX genes, respectively, whereas the numbers of identified CAT genes were four in each species. Comparative phylogenetic analysis revealed more homology among cultivated and wild soybeans relative to other legumes. Exon/intron structure, motif and synteny blocks are conserved in cultivated and wild species. According to the Ka/Ks value, purifying selection is a major force for evolution of these gene families in wild soybean; however, the APX gene family was evolved by both positive and purifying selection in cultivated soybean. Segmental duplication was a major factor involved in the expansion of APX and CAT genes. Expression patterns revealed that APX and CAT genes are differentially expressed across fourteen different soybean tissues under water deficit (WD), heat stress (HS) and combined drought plus heat stress (WD + HS). Altogether, the current study provides broad insights into these gene families in soybeans. Our results indicate that APX and CAT gene families modulate multiple stress response in soybeans.

Exogenous brassinolide improves the antioxidant capacity of Pinellia ternata by enhancing the enzymatic and nonenzymatic defense systems under non-stress conditions.

Brassinolide (BR) improves the antioxidant capacity of plants under various abiotic stresses. However, it is not clear about the effect of BR on the antioxidant capacity in plants under non-stress conditions. In the present study, the antioxidant defense response of Pinellia ternata was to be assessed by applying BR and propiconazole (Pcz) under non-stress conditions. BR treatment enhanced the flavonoid content, peroxidase, and ascorbate peroxidase (APX) activity by 12.31, 30.62, and 25.08% and led to an increase in 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity by 4.31% and a decrease in malondialdehyde content by 1.04%. Exogenous application of BR improved the expression levels of PAL, CHS, CHI, and DFR genes by 3. 18-, 3. 39-, 2. 21-, and 0.87-fold in flavonoid biosynthesis, PGI, PMI, and GME genes by 6. 60-, 1437. 79-, and 3.11-fold in ascorbic acid (ASA), biosynthesis, and γECs and GSHS genes by 6.08- and 2.61-fold in glutathione (GSH) biosynthesis pathway, and the expression of these genes were inhibited by Pcz treatment. In addition, BR treatment promoted the ASA–GSH cycle by enhancing the expression of APX, DHAR, and MDHAR genes, which were enhanced by 3. 33-, 157. 85-, and 154.91-fold, respectively. These results provided novel insights into the effect of BR on the antioxidant capacity in bulbil of P. ternata under non-stress conditions and useful knowledge of applying BR to enhance the antioxidant capacity of plants.

Assessing the effects of nickel on, e.g., Medicago sativa L. nodules using multidisciplinary approach.

Industrial wastes and fertilizers can introduce excessive levels of nickel (Ni) into the environment, potentially causing threats to plants, animals, as well as human beings. However, the number of studies on the effects of Ni toxicity on nodules is fairly limited. To address this issue, the effects of increasing Ni concentration on alfalfa nodules were assessed at chemical, biochemical, and transcriptomic levels. For this purpose, plants were grown in soils supplied with Ni (control, 0 mg/kg; C1, 50 mg/kg; C2, 150 mg/kg; C3, 250 mg/kg; and C4, 500 mg/kg) for 90 days. Ni loads in leaves, roots, and nodules were monitored after the exposure period. A set of biochemical biomarkers of oxidative stress was determined in nodules including antioxidants and metal homeostasis as well as lipid peroxidation. Gene expression levels of the main targets involved in oxidative stress and metal homeostasis were assessed. Our data indicated a high concentration of Ni in leaves, roots, and nodules where values reached 25.64 ± 3.04 mg/kg, 83.23 ± 5.16 mg/kg, and 125.71 ± 4.53 mg/kg in dry weight, respectively. Moreover, a significant increase in nodule biomass was observed in plants exposed to C4 in comparison to control treatment and percentage increased by 63%. Then, lipid peroxidation increased with a rate of 95% in nodules exposed to C4. Enzymatic activities were enhanced remarkably, suggesting the occurrence of oxidative stress, with increased superoxide dismutase (SOD), glutathione reductase (GR), and ascorbate peroxidase (APX). Our results showed also a significant upregulation of SOD, GR and APX genes in nodules. Nodule homoglutathione (HGSH) levels increased with the different Ni concentrations, with a remarkable decrease of glutathione S-transferase (GST) activity and glutathione (GSH) content for the highest Ni concentration with 43% and 52% reduction, respectively. The phytochelatin (PC) and metallothionein (MT) concentrations increased in nodules, which implied the triggering of a cellular protection mechanism for coping with Ni toxicity. The results suggested that Ni promotes a drastic oxidative stress in alfalfa nodules, yet the expression of MT and PC to reduce Ni toxicity could be used as Ni stress bioindicators. Our findings provide new insights into the central role of alfalfa nodules in limiting the harmful effects of soil pollution. Therefore, nodules co-expressing antioxidant enzymes may have high phytoremediation potential.

Molecular and Physiological Responses of Citrus sinensis Leaves to Long-Term Low pH Revealed by RNA-Seq Integrated with Targeted Metabolomics.

Low pH-induced alterations in gene expression profiles and organic acids (OA) and free amino acid (FAA) abundances were investigated in sweet orange [Citrus sinensis (L.) Osbeck cv. Xuegan] leaves. We identified 503 downregulated and 349 upregulated genes in low pH-treated leaves. Further analysis indicated that low pH impaired light reaction and carbon fixation in photosynthetic organisms, thereby lowering photosynthesis in leaves. Low pH reduced carbon and carbohydrate metabolisms, OA biosynthesis and ATP production in leaves. Low pH downregulated the biosynthesis of nitrogen compounds, proteins, and FAAs in leaves, which might be conducive to maintaining energy homeostasis during ATP deprivation. Low pH-treated leaves displayed some adaptive responses to phosphate starvation, including phosphate recycling, lipid remodeling, and phosphate transport, thus enhancing leaf acid-tolerance. Low pH upregulated the expression of some reactive oxygen species (ROS) and aldehyde detoxifying enzyme (peroxidase and superoxidase) genes and the concentrations of some antioxidants (L-tryptophan, L-proline, nicotinic acid, pantothenic acid, and pyroglutamic acid), but it impaired the pentose phosphate pathway and VE and secondary metabolite biosynthesis and downregulated the expression of some ROS and aldehyde detoxifying enzyme (ascorbate peroxidase, aldo-keto reductase, and 2-alkenal reductase) genes and the concentrations of some antioxidants (pyridoxine and γ-aminobutyric acid), thus disturbing the balance between production and detoxification of ROS and aldehydes and causing oxidative damage to leaves.

Small HSPs play an important role in crosstalk between HSF-HSP and ROS pathways in heat stress response through transcriptomic analysis in lilies (Lilium longiflorum)

BackgroundHigh temperature seriously limits the annual production of fresh cut lilies, which is one of the four major cut flowers in the global cut flower market. There were few transcriptomes focused on the gene expression of lilies under heat stress. In order to reveal the potential heat response patterns in bulbous plants and provide important genes for further genetic engineering techniques to improve thermotolerance of lily, RNA sequencing of lilies under heat treatments were conducted.ResultsIn this study, seedlings of Lilium longiflorum ‘White Heaven’ were heat-treated at 37 °C for different lengths of time (0 h, 0.5 h, 1 h, 3 h, 6 h, and 12 h with a 12 h-light/12 h-dark cycle). The leaves of these lily seedlings were immediately collected after heat treatments and quickly put into liquid nitrogen for RNA sequencing. 109,364,486–171,487,430 clean reads and 55,044 unigenes including 21,608 differentially expressed genes (DEGs) (fold change ≥2) were obtained after heat treatment. The number of DEGs increased sharply during the heat treatments of 0.5 h–1 h and 1 h–3 h compared to that of other periods. Genes of the heat stress transcription factor (HSF) family and the small heat shock proteins (small HSPs, also known as HSP20) family responded to heat stress early and quickly. Compared to that of the calcium signal and hormone pathways, DEGs of the HSF-HSP pathway and reactive oxygen species (ROS) pathway were significantly and highly induced. Moreover, they had the similar expression pattern in response to heat stress. Small HSPs family genes were the major components in the 50 most highly induced genes at each heat stress treatment and involved in ROS pathway in the rapid response to heat stress. Furthermore, the barley stripe mosaic virus induced gene silencing (BSMV-VIGS) of LlHsfA2 caused a significantly reduced thermotolerance phenotype in Lilium longiflorum ‘White Heaven’, meanwhile decreasing the expression of small HSPs family genes and increasing the ROS scavenging enzyme ascorbate peroxidase (APX) genes, indicating the potential interplay between these two pathways.ConclusionsBased on our transcriptomic analysis, we provide a new finding that small HSPs play important roles in crosstalk between HSF-HSP and ROS pathways in heat stress response of lily, which also supply the groundwork for understanding the mechanism of heat stress in bulbous plants.