Penicillium sp. was cultured using One Strain Many Compounds (OSMAC) conditions as a model system to demonstrate how this strategy can help in understanding metabolic profiles and determining bioactive metabolites with the application of metabolomics and statistical analyses, as well as molecular networking. Penicillium sp. was fermented in different culture media and the crude extracts from mycelial biomass (CEm) and broth (CEb) were obtained, evaluated against bacterial strains (Staphylococcus aureus and Pseudomonas aeruginosa), and the metabolomic profiles by LC-DAD-MS were obtained and chemometrics statistical analyses were applied. The CEm and CEb extracts presented different chemical profiles and antibacterial activities; the highest activities observed were against S. aureus from CEm . The antibacterial properties from the extracts were impacted for culture media from which the strain was fermented. From the Volcano plot analysis, it was possible to determine statistically the most relevant features for the antibacterial activity, which were also confirmed from biplots of PCA as strong features for the bioactive extracts. These compounds included 75 (13-oxoverruculogen isomer), 78 , 87 , 88 (helvamide), 92 (viridicatumtoxin A), 96 , 101 , 106 , 110 , and 112 (pre-viridicatumtoxin). Thus, these features included diketopiperazines, meroterpenoids, and polyketides, such as indole alkaloids, austalides, and viridicatumtoxin A, a rare tetracycline.Metabolomics strategies are important tools to get holistic chemical information from a system, but they are scarcely applied to endophytic fungi to understand their chemical profiles of biosynthesized metabolites. Here  The search for natural products with biological activities from endophytic fungi is imminent, due to their remarkable ability to biosynthesize compounds with high levels of structural complexity and prominent pharmacological properties ,2. EndopThe regulation of the biosynthesis of natural products is complex and the conditions under which the microorganisms are cultivated can drastically influence the production of metabolites . The appMetabolomics can be defined as a comprehensive analysis of metabolites to get the metabolic information of low weight <1000 Da) from an organism or biological system . The met00 Da froPenicillium sp. 5MP2F4 applying the OSMAC strategy and screening the antibacterial and antibiofilm activities. Additionally, we aimed to get broad chemical information by metabolomics and chemometrics tools.Given that there is still no consensus for an initial and wide investigation of metabolite production capacity by endophytic fungi and that the choice of culture medium influences this biosynthetic production from the microorganisms, here we aimed to evaluate the metabolites produced by the strain Penicillium sp. 5MP2F4 , a bromeliad species occurring in the ironstone outcrops of the Pantanal biome in Brazil. The fungal strain was identified according to morphological traits and ITS rDNA sequence analysis, as reported by Cassemiro et al. + (C27H32N3O6+), relative to the dehydration reaction in the ionization source. The fragment ion of 85 at m/z 352 (C19H18N3O4+) was yielded from the cleavage of the eight-membered ring. Subsequent cleavage of the piperazinodione ring produces the product ion at m/z 255.0766 (C14H11N2O3+) after the loss of C5H7NO and with subsequent loss of a CO molecule (28 u) producing the fragment at m/z 227.0808 [C13H11N2O2]+, similar to what was described in the literature pyrazine-1,4-dione] yielded an intense product ion at m/z 226.1227 (C15H16NO+). An additional cleavage of the central ring resulted in the formation of the ion of m/z 212.105 (C14H14NO+), and the fragment ion m/z 255.1476 (C16H19N2O+) was yielded from the cleavage of the piperazinodione ring, and subsequently, the loss of 2-methylpropene yielded the ion at m/z 199.0868 (C12H11N2O+). This fragmentation profile is compatible with fumitremorgin C (56) described by Fornal et al. +, 338 [M+H-C4H8-H2O]+, and 310 [M+H-C4H8-H2O-CO]+ from 22. Thus, they were annotated as dihydroxyfumitremorgin C/cyclotryprostatin A isomers.The compounds 94 showed an intense ion at m/z 462.2389 [M+H-H2O]+, compatible with the molecular formula C27H33N3O5, and relative to the dehydration reaction in the ionization source of the mass spectrometer. This intense dehydration reaction is common when there are OH substituents on the central ring from secondary and/or tertiary aliphatic alcohols, such as for the compounds 94 and 90. The product ion of m/z 394, observed from m/z 462 (94), was yielded by the loss of an isoprenic unit , and subsequent elimination of the pyrrole ring produced the ion m/z 327. A similar fragmentation was observed for 86, 89–91, and 94, which were annotated as fumitremorgin B isomers , deoxy-fumitremorgin B (89), and hydroxy-fumitremorgin B (90). Additionally, the compounds oxofumitremorgin B (84) and di-deoxy-fumitremorgin B (99) were also annotated. Several compounds annotated here are diketopiperazine-type indolic alkaloids, which have been described as precursors of the verruculogen biosynthetic pathway, as observed in The metabolite 54 and 64 revealed the same molecular formula, C21H25N3O2, and a similar fragmentation profile; moreover, 54 showed a score hit with tryprostatin B deposited in the GNPS. The product ions at m/z 296 [M+H-C4H8]+ and 268 [M+H-C5H8]+ indicated the presence of an isoprene group and yielded fragments with subsequent losses of CO (28 u) to produce the ions m/z 268 and 240. The peaks 54 and 64 were annotated as tryprostatin B (isomers). Moreover, the metabolites 26, 28, 30, 38, 39, 40, 43, and 48 were annotated as dihydroxy-spirotryprostatin A, spirotryprostatin G, cyclotryprostatin E, cyclotryprostatin C or hydroxycycloprostatin A, cyclotryprostatin B, cclotryprostatin C, oxo-cyclotryprostatin A, and dehydroxycyclotryprostatin C, respectively.Compounds 44, 50, 63, 69, 73, 78, 87, 96, 101, and 106 , 50 , 63 , 69 , 73 , 78 , 87 , 96 , 101 , and 106  (m/z 207 (C11H11O4+) and 177 (C10H9O3+). The fragment ion m/z 207 is yielded from retro Diels-Alder fission and subsequent loss of CH2O (30 u) that indicates the presence of the lactone group (u) indicated the presence of non-phenolic hydroxyls, such as those observed for the metabolites 44 (m/z 445 [M+H-H2O]+ and 409 [M+H-3xH2O]+) and 69 (m/z 459 [M+H-H2O]+ and 441 [M+H-2xH2O]+). Thus, these consecutive losses of water molecules were important in suggesting the aliphatic hydroxyl groups in the austalides, as well as the losses to yield the acylium fragment .It was possible to observe another cluster, formed by austalide-type meroterpenoids, which includes the compounds  and 106 . These mstalides . The com25H32O5)  revealedne group . For allO-desmethyl austalide B (50) was confirmed by the authentic standard. Beyond the similar fragmentation pathway described here, the metabolite 63 exhibited a cosine of 0.94 and 0.96 between the austalides 69 and 44, revealing greater structural similarity between them. The metabolite 73 showed a cosine of 0.89 in relation to 87 and 101. Thus, 63 and 73 were annotated as austalide Q acid and austalide J or 13-O-desacetyl austalide I, respectively. Metabolites 101  and 106  showed a fragmentation profile similar to that observed for austalides and the cosine between them was 0.97, suggesting a high similarity between them and a difference of 2 u relative to the presence of a double bond. They were annotated as dihydroaustalide K (101) and austalide K (106).The compound 17-92, 110, and 112 showed two absorption bands, at λmax ≈ 285 and 430 nm in their UV spectra. Their deprotonated ions m/z 564.1870, 563.1918, and 566.2027 are compatible with molecular formulas C30H31NO10, C31H32O10 e C30H33NO10, respectively. These compounds exhibited polyketide fragmentation patterns [u), and for 92 and 112 an additional loss of an ammonia molecule (17 u) was also observed. Compound 92 was also confirmed by injection of authentic standards. Thus, 92, 110, and 112 were annotated as viridicatumtoxin A, spirohexalin, and previridicatum , 113 , 116 , and 120 . These compounds were annotated as isomers of lysodiacylglyceryltrimethylhomoserine (LDGTS)  and diacylglyceryl trimethylhomoserine (DGTS) (120). Additionally, a cluster with phospholipids was observed, and it was generated by compounds 95 , 98 , 104 , 107 , and 109 .Lipid classes were also detected in the molecular network . A clustN-acetyl brasiliamide I (34), oxo-cyclotryprostatin A (43), chrysosporazine P (61), chrysosporazine D (81), helvamide (88), and oxoverrucologen isomers  are described here for the first time in the genus Penicillium, while the metabolites viridicatumtoxin A (92), spirohexaline (110), previridicatumtoxin (112), austalide H acid (44), 17-O-desmethyl austalide B (50), austalide Q acid (63), austalide H (69), austalide J/13-O-desacety l austalide (73), austalide P acid (78), austalide L/W (87), austalide P (96), dihydroaustalide K (101), and austalide k (106) are described for the first time from a microorganism isolated in Brazil. Therefore, our study contributes to enlarging the chemical knowledge from the genus Penicillium and Brazilian microorganisms.The compounds Penicillium sp. was applied to evaluate the effects of five different culture media on the metabolome of strain 5MP2F4.Fungi can biosynthesize diverse structures with different polarities, functions, and biological properties , and they can occur at different levels of abundance in each microorganism ,6,25,31.Bromelia balansae Mez that occurs in ironstone outcrops in the Pantanal biome of Mato Grosso do Sul, Brazil [This strain was collected as an endophyte from , Brazil . This pl, Brazil . In addi, Brazil ,36,37,38The OSMAC strategy has been applied to obtain a greater diversity of microbial metabolites as well as in the search for new bioactive compounds . AlthougPenicillium sp are dependent on the culture medium, since the exclusive metabolites were observed from a specific culture medium, as were differences in their intensities. These appointments have been described from strains of Penicillium with the application of the OSMAC strategy [Our efforts here involved the description of a great variability of metabolites produced by an endophytic fungus and the annotation of them from both broth and mycelial biomass, which revealed different chemical profiles. We also observed that the metabolites biosynthesized by strain strategy ,42, but From the statistical analyses illustrated in Penicillium sp. 5MP2F4 were highlighted here as chemical classes related to antibacterial activity against S. aureus.The metabolomics data were also correlated with the antibacterial properties of the extracts (CEb and CEm obtained by OSMAC strategy) by statistical analyses to suggest the possible bioactive metabolites, optimizing the targets for further studies. The application of metabolomics and statistical tools to accelerate the research of bioactive compounds has been used by several studies, including compounds with antibacterial, larvicidal, and antileishmanial properties ,43,44,45S. aureus bacterial growth. The increase in the number of bacterial infections together with the shortage of new antibiotics is a huge public health problem. It is estimated that the increase in the rates of bacterial infections caused by microorganisms ranks among the main causes of human mortality, and the bacteria S. aureus is on the list of multiresistant bacteria that call for the development of new drugs [The extracts from broth and biomass (CEb and CEm) showed different antibacterial activities and promising results to inhibit ew drugs . In thisew drugs , highligew drugs .In this work, we annotated different types of diketopiperazine-type alkaloids, such as brasiliamides A, C, D, E, I, and N-acetyl brasiliamide I. They are a class of metabolites widely distributed in fungi, bacteria, sponges, and actinomycetes. Brasiliamides are alkaloids that contain a piperazine grouping and they possess diverse biological activities including antibiotic, immunosuppressive, and antineoplastic activities, and they have great potential in future therapeutic applications . Their bPenicillium brasilianum Batista JV-379, and their potential to produce convulsions in animals was evaluated, using the silkworm (Bombys mori) as an experimental model. Brasiliamide A and B showed ED50 of 300 and 50 µg/g, , respectively. In this work, these brasiliamides were proposed as a new class of tremorgenic substances [The brasiliamides A and B  were firbstances . Later, bstances .Penicillium brasilianum, an endophyte of Melia azedarach (Meliaceae). The antimicrobial activity of brasiliamide A against a strain of Bacillus subtilis was also described, revealing an MIC of 250 µg/mL [P. brasilianum WZXY-M122-9 was used to study the biosynthesis of brasiliamides and the implication of its enzymes. Thus, the authors described for the first time brasiliamide I [N-acetyl brasiliamide I (34) was also annotated, which has not yet been reported in the literature.Fill and co-workers described the isolation of brasiliamides A-F from 50 µg/mL , and thi91), oxoverruculogen (isomer) (75), hydroxy-verruculogen (83), deoxy-fumitremorgin B (89), di-deoxy-fumitremorgin B (99) are also compounds statically related to the antibacterial activity. These compounds are diketopiperazine-type indolic alkaloids that are commonly reported from Aspergillus and Penicillium species [56) has been described as a potent inhibitor of breast cancer resistance protein [85), a tremorgenic mycotoxin, acts in the central nervous system and induces sustained tremors, convulsions, and can lead to the death of animals. Furthermore, verruculogen inhibits the high conductance calcium-activated potassium channels of smooth muscle, and it has been a target of further drug research [Fumitrermorgin B (isomer) , austalide P acid (78), austalide L/austalide W (87), austalide P (96), dihydroaustalide K (101), and austalide K (106). They are meroterpenoids with several activities described for them, such as antitumor, anti-inflammatory, antimicrobial, antiviral (influenza A H1N1), and osteoclast differentiation inhibitors [Halomonas aquamarina, Polaribacter irgensii, Pseudoalteromonas elyakovii, Roseobacter litoralis, Shewanella putrefaciens, Vibrio harveyi, V. natriegens and V. carchariae), and they showed a broad spectrum of activity with MIC values between 0.001 and 1 µg/mL [S. aureus and Escherichia coli, but they were not active [Aspergillus sp. isolated from the sponge Tethya aurantium, and they are suggested as antibacterial compounds useful in aquaculture [Austalides also presented correlations with antibacterial activity, including austalide Q acid (hibitors . Althoug 1 µg/mL . In addit active . These aaculture .92), spirohexaline (110), and previridicatumtoxin (112) are recognized by their antibacterial potentials [The tetracycline-like polyketides, such as the annotated compounds viridicatumtoxin A  is a substance with a hybrid polyketide-isoprenoid structure. It is a rare tetracycline-like compound produced by fungi, which is structurally considered privileged because it possesses the ability to interact with different biological targets [S. aureus with MIC of 0.5 µg/mL [Viridicatumtoxin A  and mycelial biomass (CEm), including differences in the chemical profiles between them and their antibacterial activities. Several metabolites annotated here are reported for the first time from an endophytic fungus of Brazil and in the genus Penicillium. The OSMAC strategy induced the biosynthesis of different compounds and the application of metabolomics and chemometrics tools were useful in indicating antibacterial compounds, such as diketopiperazines, meroterpenoids, and polyketides, highlighting the rare tetracyclin viridicatumtoxin A. These findings reveal that the chemical and biological study of simple cultures can be useful in optimizing the knowledge of the metabolic potential of fungal strains.From the endophytic fungi


Haemonchus contortus is the most prevalent and virulent, resulting in health problems and production losses. Therefore, selecting sheep resistant to H. contortus is a suitable and sustainable strategy for controlling endoparasites in flocks. Here, 287 lambs of the native Brazilian Morada Nova hair sheep breed were subjected to two consecutive artificial infections with H. contortus and assessed for fecal egg count (FEC), packed cell volume (PCV), and live weight (LW). Forty-four animals ranked as having extreme resistance phenotypes were genotyped using the Illumina OvineSNP50v3 chip. A case–control genome-wide association study (GWAS) detected 37 significant (p < 0.001) markers in 12 ovine chromosomes in regions harboring quantitative trait loci (QTL) for FEC, Trichostrongylus spp. adults and larvae, weight, and fat; and candidate genes for immune responses, mucins, hematological parameters, homeostasis, and growth. Four single-nucleotide polymorphisms  genotyped by qPCR followed by high-resolution melting (HRM) were associated with FEC and LW. Therefore, molecular markers detected by GWAS for H. contortus resistance in Morada Nova sheep may support animal selection programs aimed at controlling gastrointestinal nematode infections in flocks. Furthermore, genotyping of candidate genes using HRM qPCR may provide a rapid and efficient tool for animal identification.Among the gastrointestinal nematodes affecting sheep,  Morada Nova is a hair sheep breed characterized by high heat tolerance and adaptation to tropical climatic conditions . TraditiHaemonchus contortus, is the most prevalent and virulent nematode species that affects small ruminants in tropical regions [H. contortus is a blood-sucking parasite of the abomasum, which causes anemia, weight loss, and death [The barber pole worm,  regions . H. contnd death . Considend death ,6, parasnd death , as it rnd death ,9.Several gene polymorphisms associated with resistance to gastrointestinal nematodes have been detected in Morada Nova sheep by candidate gene genotyping ,11; howeH. contortus-resistant and -susceptible Morada Nova sheep was performed using a case–control design to identify genomic regions involved in resistance to gastrointestinal nematodes. Furthermore, to our knowledge, this was the first GWAS related to H. contortus resistance in a Brazilian native parasite-resistant Morada Nova sheep breed resulting in 37 significant markers located on 12 ovine chromosomes harboring regions previously associated with resistance-related traits and functional candidate genes. In addition to providing guidance for studies in other breeds, knowledge of the molecular mechanisms involved in parasite resistance will support the use and maintenance of Morada Nova sheep as a genetic stock for gene introgression and direct use in production systems, aiming to control nematode infections in flocks.Therefore, a GWAS of p < 0.001) single nucleotide polymorphism (SNP) markers located on 12 ovine chromosomes  (The genome-wide association study (GWAS) identified 37 significant ( and 20) .Using Haploview, linkage disequilibrium (LD) blocks were detected for markers located in OAR 2 and 8, and strong LD was observed for markers in OAR 7, 11, and 18, .Nematodirus, H. contortus, and Trichostrongylus colubriformis fecal egg counts (FEC), larvae and adults of Trichostrongylus spp. in the abomasum and small intestine, weight, and fat traits , CD96 (1:174690709–174790572), CD200 (1:175479877–175498685), CFI (6:15708801–15753015), DPP4 (2:146632572–146713094), FER (5:105044035–105461672), LEF1 (6:17197574–17304587), NFE2L2 (2:131754249–131759072), and the TREML1, TREM1, and TREM2 gene cluster (20:15158837–15237353);Immune response: -GALNT10 (5:63016723–63244040), GCNT3 (7:47666417–47667739), and PGC (20:15636993–15649730);Gastric mucosa and mucins: -CLEC14A (18:47277443–47278737), DLX1 (2:136599373–136602848), EDNRA (17:10416386–10482226), and SH2B3 (17:54732694–54757389);Hematological parameters: -CCDC80 (1:175729335–175762809), HS3ST5 (8:23301492–23469323), and PLA2G1B (17:62282828–62288877);Homeostasis: -DPPA2 (1:172525876–172536184), HOXD gene cluster (2:132820016–132915560), KLHL41 (2:138842626–138856360), MYF5 (3:116620277–116622399), and PITX2 (6:14934427–14955991);Growth and muscle development: -MAPK7 (11:33629196–33632861) and SREBF1 (11:34176887–34191779).Lipid metabolism and fat deposition: The biological roles of genes  potentially involved in nematode resistance in Morada Nova sheep  are relaReal-time polymerase chain reaction (qPCR) followed by high-resolution melting (HRM) analyses confirmed the genotypes attributed using the chip in all 44 animals for OAR1_rs427671974, OAR2_rs419988472, OAR5_rs424070217, and OAR17_rs401006318, demonstrating 100% agreement. In addition, these four SNPs were associated with FEC A, and OAH. contortus resistance using OvineSNP50v3 chip genotyping of 44 ranked phenotyped  animals from a population of 287 Morada Nova sheep, with data from two experimental parasite challenges. GWAS detected 37 significant molecular markers, some in strong LD, on 12 ovine chromosomes in genomic regions harboring several functional candidate genes and QTLs for related traits. GWAS was validated by HRM qPCR genotyping of four significant SNP markers that were associated with FEC and LW. Mixed gastrointestinal parasite species are present under natural or field infection [H. contortus and repetitive phenotype collection, may have contributed to the suitability of the employed GWAS approach.The present study employed a case–control GWAS for nfection . Then, tH. contortus, T. colubriformis, and Nematodirus FEC and Trichostrongylus spp. adults and larvae in the abomasum and small intestine, in OAR 2, 3, 7, 8, 11, 17, and 18. In addition, superposition of production trait QTLs, such as weight and fat, was detected in OAR 1, 2, 3, 5, 6, 8, 11, and 18. An association with production traits was expected in the present study, as live weight was used to rank animals with extreme phenotypes. However, several studies based solely on FEC and nematode infection rates have also detected regions spanning production QTLs, suggesting a correlation between live weight and growth with resistance [H. contortus resistance in Morada Nova sheep did not harbor reported QTLs for related traits, as observed for OAR 5, 15, 17, and 20, confirming that different molecular mechanisms may regulate resistance in different breeds. Consequently, due to the genetically fragmented nature of sheep and goat populations, GWAS information derived from one breed cannot be extrapolated to others without proper validation [Significant markers were located in genomic regions harboring QTLs for parasite resistance, such as sistance , as wellsistance . Some geH. contortus infection [Regarding the enriched terms detected in functional analyses, homeobox has previously been associated with adaptive immune response in cattle , and thenfection . H. contortus secretes and excretes antigens that stimulate host inflammatory, humoral, and cellular immune responses. Consequently, the recruited T-helper cells release cytokines, mainly interleukins, which activate IgE synthesis, eosinophils, mast cells, and globular leukocytes in the mucosa. These events are followed by B-cell activation and antibody production (IgA and IgG1). Additionally, mast cell and eosinophil inflammatory products, such as histamines, proteases, leukotrienes, and prostaglandins, lead to mucus production and smooth muscle contraction, which induce parasitic paralysis, elimination, or death. Furthermore, events favoring host homeostasis and coping with parasitic loads, such as protein and energy metabolism, hematological parameters, and body weight, have also been associated with resistance and/or resilience to gastrointestinal nematodes [Several mechanisms can disrupt parasite establishment and confer host resistance to gastrointestinal nematodes ,19. Brieematodes ,20. BaseBTLA, an immunoglobulin superfamily member, and LEF1, an enhancer of T-cell receptor-alpha, was increased in peripheral blood mononuclear cells in Suffolk sheep following exposure to H. contortus larvae antigens [BTLA expression in host T CD4(+) cells and innate leukocytes affected intestinal immunity and Strongyloides ratti infection [CFI, which regulates the complement cascade, was detected in the abomasum of H. contortus-resistant sheep breeds [CFI was also associated with H. contortus FEC [CD96, an immunoglobulin superfamily member, has been detected in nematode-susceptible goats [CD200 [NFE2L2 (or Nrf2) [Leishmania infection. TREML1 (or TLT-1), TREM2, and TREM1 genes, which are involved in inflammatory, innate, and adaptive immune responses, were associated with tick resistance in cattle [TRML1 in clot formation and inflammatory or immune-induced bleeding has been reported [TREM1 was detected in sheep peripheral blood mononuclear cells during chronic infection with Fasciola hepatica [FER, a tyrosine kinase involved in leukocyte recruitment, regulated the intestinal epithelial lipopolysaccharide barrier in response to bacteria [DPP4, which is involved in metabolism and immune regulation, was associated with the innate immune response to virus [Among candidate genes related to the immune response, the expression of antigens . In addinfection . Increasp breeds ,24,25, artus FEC . Increasle goats  and cattle goats . CD200 [s [CD200 , a glycoor Nrf2) , which in cattle , and thereported ,33. Furthepatica . FER, a bacteria . DPP4, wto virus  and demoto virus .GALNT10 gene, which drives mucin-type O-glycan synthesis, is a paralog of GALNTL6, which was found by GWAS to be associated with gastrointestinal parasite resistance in sheep [PGC, a component of the gastric mucosa, has been detected in the abomasum of a sheep breed resistant to H. contortus [GCNT3, which plays a role in mucin-type glycoproteins, has been detected in the small intestine in response to Cooperia oncophora [Ostertagia ostertagi infection [Regarding the gastric mucosa and mucins, the in sheep . In addiontortus . In cattncophora  and in tnfection . H. contortus is a hematophagous parasite, genes affecting hematological parameters in hosts may have a potential role in resistance. DLX1, a homeobox transcription factor, regulated the TGF-β superfamily during blood production [EDNRA, which encodes the receptor for endothelin-1, affected vasoconstriction in yaks [CLEC14A controlled angiogenesis in mice [SH2B3, a negative regulator of cytokine signaling, was found to be associated with erythrocyte traits in sheep by GWAS [Considering that oduction ; EDNRA,  in yaks ; CLEC14A in mice ; and SH2 by GWAS . PLA2G1B, a phospholipase A2 that regulates energy metabolism and inflammation in the intestine, was considered an endogenous anthelmintic that induces Heligmosomoides polygyrus and Nippostrongylous brasiliensis death in mice [HS3ST5, a cell-surface heparan sulfate, acted as a receptor facilitating Trypanosoma cruzi [Toxoplasma gondii [CCDC80, which enables glycosaminoglycan binding activity, was affected by Trypanosoma cruzi infection [Regarding homeostasis,  in mice . HS3ST5,ma cruzi  and Toxoa gondii  invasionnfection . HOXD1, HOXD3, HOXD10, HOXD12, and HOXD13 have been detected by GWAS for muscularity in cattle [PITX2 was related to growth in sheep [DPPA2 was involved in myogenesis [KLHL41 was associated with skeletal muscle differentiation [MYF5, a myogenic factor, was associated with growth in sheep [MAPK7 was involved in adipocyte differentiation [SREBF1 affected fat metabolism and deposition in sheep [Some of the identified genes in this study were associated with growth, muscle development, and fat traits. Homeobox genes n cattle , and PITin sheep  and weigin sheep . DPPA2 wogenesis , KLHL41 ntiation , and MYFin sheep . Furtherntiation , and SREin sheep . H. contortus resistance in Morada Nova sheep was validated by association analyses of four significant SNP markers genotyped by HRM qPCR with FEC and LW in animals. No association with PCV was detected. However, while FEC and LW were used as factors multiplied by 0.4 to rank animals in extreme phenotypes, PCV presented a lower weight (0.2) in the equation. This decision was based on the fact that PCV presented lower variability among animals compared with FEC and LW, suggesting that Morada Nova sheep are resilient, rather than fully resistant to H. contortus [The analytical strategy employed by GWAS to detect ontortus . The BB For phenotypic evaluation, 2 g of feces was collected from the rectum, mixed with 28 mL of sodium chloride-saturated solution, and evaluated in a McMaster chamber . The tot®) to remove natural infection with gastrointestinal nematodes. After two FEC at 7-day intervals, animals were experimentally infected with 4000 H. contortus third-stage larvae (L3) from an anthelmintic susceptible isolate [3H. contortus of the same isolate, followed by the sampling protocol described previously. At 42 DPI of the second parasitic challenge, mean FEC, PCV, and LW were calculated from the collected data. Subsequently, animals were ranked as extreme phenotypes (extremely resistant and extremely susceptible to H. contortus) based on the following equation:The DNA samples and phenotypic data  used to rank animals with extreme phenotypes were obtained in 2017 and 2018 . Briefly isolate . FEC wasBased on phenotype ranking, 44 lambs were selected considering homogeneous phenotype distribution (21 resistant and 23 susceptible), sex , and number of progenies (7–12) from four rams with a large number of descendants. This resulted in 12 resistant females, 11 susceptible females, 9 resistant males, and 12 susceptible males. In addition, differences in rank mean value varied from 8 to 36 times between extreme phenotype progenies from each ram. DNA was extracted by saline precipitation  from ven−15. An efficient mixed-model association expedited (EMMAX) was used to identify nominal p-values for phenotype–genotype interactions, and then PLINK  was used for Bonferroni testing. Linkage disequilibrium (LD) between SNP markers was calculated using Haploview [Genotyping chip data were subjected to quality control filtering, with 47,782 single-nucleotide polymorphisms (SNP) retained for analyses by case–control GWAS . PLINK waploview . www.ensembl.org/biomart accessed on 28 April 2022) and the Ensembl Genes 106 database. The same genomic interval was investigated for superposition with related sheep quantitative trait loci (QTL) mapped on SheepQTLdb  using the Sheep Genome Track on OAR_3.1 . Genes located within a 2 Mbp window upstream and downstream of each marker were searched in the OAR v3.1 sheep genome using BioMart (https://david.ncifcrf.gov/home.jsp accessed on 29 April 2022) with Ovis aries genome as the background. DAVID was used to identify the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways [p = 0.05 in Fisher’s exact test [https://www.genecards.org/ accessed on 9 May 2022) and a literature review were used to identify functional candidate genes. ENSEMBL_Gene_ID was used as a gene list in the functional annotation tool of the DAVID Bioinformatics Resources 2021 update  located on four ovine chromosomes , were genotyped via quantitative PCR (qPCR) followed by high-resolution melting (HRM) in the 44 lambs subjected to GWAS. The HRM qPCR assay consisted of 1X SsoFast Evagreen Supermix (Bio-Rad 172–5200), 0.3 µM of each primer , and 5 nThe effects of SNPs on FEC, PCV, and LW were analyzed as described by . BrieflyH. contortus resistance profiles using 44 extreme phenotype Morada Nova sheep genotyped by Illumina OvineSNP50 chip resulted in 37 significant markers in OAR 1, 2, 3, 5, 6, 7, 8, 11, 15, 17, 18, and 20. Marker genomic regions harbored QTLs for FEC, adults and larvae of Trichostrongylus spp. in the abomasum and small intestine, weight, and fat traits, in addition to functional candidate genes related to the immune response, gastric mucin, hematological parameters, homeostasis, growth, and fat deposition. HRM qPCR genotyping of four molecular markers  validated their association with FEC and live weight. Thus, the obtained results can be used for the selection of native Morada Nova sheep with the aim of controlling gastrointestinal nematode infection in flocks and increasing parasite resistance in production systems.GWAS to determine


Oxidative stress and endothelial dysfunction have been shown to play crucial roles in the pathophysiology of COVID-19 (coronavirus disease 2019). On these grounds, we sought to investigate the impact of COVID-19 on lipid peroxidation and ferroptosis in human endothelial cells. We hypothesized that oxidative stress and lipid peroxidation induced by COVID-19 in endothelial cells could be linked to the disease outcome. Thus, we collected serum from COVID-19 patients on hospital admission, and we incubated these sera with human endothelial cells, comparing the effects on the generation of reactive oxygen species (ROS) and lipid peroxidation between patients who survived and patients who did not survive. We found that the serum from non-survivors significantly increased lipid peroxidation. Moreover, serum from non-survivors markedly regulated the expression levels of the main markers of ferroptosis, including GPX4, SLC7A11, FTH1, and SAT1, a response that was rescued by silencing TNFR1 on endothelial cells. Taken together, our data indicate that serum from patients who did not survive COVID-19 triggers lipid peroxidation in human endothelial cells. We and others have evidenced the fundamental role of endothelial cells in the pathobiology of the coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus (SARS-CoV-2) ,15,16,17The overproduction of reactive oxygen species (ROS) is known to induce endothelial dysfunction through different mechanisms, including ferroptosis ,50, a noWe hypothesized that oxidative stress and subsequent lipid peroxidation induced by COVID-19 in endothelial cells could be linked with the disease outcome. To test this hypothesis, we collected serum from COVID-19 patients on hospital admission, and we incubated these sera with human endothelial cells, comparing the effects on ROS generation and lipid peroxidation between patients who survived and patients who did not survive.p < 0.0001) increased production of both cellular  of serum obtained from patients who did not survive COVID-19, comparing these effects with the serum of patients who did survive. The main characteristics of the patients are shown in cellular  and mitocellular  ROS in HOxidative stress has been linked to lipid peroxidation, a process that has been recently suggested to play a critical role in COVID-19 ; on thesSince the selenoprotein glutathione peroxidase 4 (GPX4) has been shown to be a major modulator of lipid peroxidation ,61,62, wc−) [1-acetyltransferase 1 (SAT1) [Increased lipid peroxidation and reduced levels of GPX4 represent two hallmarks of ferroptosis ,64; thusc−) ,68,69,70c−) ,72,73, w1 (SAT1) ,75,76 wap < 0.001). Several studies have consistently shown that increased TNFα serum levels on hospital admission are associated with the risk of mortality in COVID-19 patients ,78,79. SThen, to mechanistically prove that TNFα was involved in the COVID-19 induced ferroptotic response, we measured by immunoblot GPX4, SLC7A11, FTH1, and SAT1 after having treated HUVECs with a siRNA for TNFR1 or scramble , which has been recently reported both as a suppressor and as an inductor of ferroptosis in a series of conflicting data ,139.2, in EGM-2 medium , as we described [2).All reagents were purchased from Merck , unless otherwise stated. HUVECs were obtained from ThermoFisher Scientific . Cells were cultured in a standard humidified atmosphere (37 °C) containing 5% COescribed ,47,140. #202011756). Normal human serum was purchased from ThermoFisher Scientific. The absence of SARS-CoV-2 in the serum was confirmed by RT-qPCR. TNFα was quantified via ELISA  following the manufacturer’s protocol.The sera from fully de-identified COVID-19 patients were provided by the Montefiore–Einstein COVID-19 biorepository; the protocol was approved by the Institutional Ethical Committee , as described previously [3 10 mM, MgCl2 2.5 mM, CaCl2 2.5 mM, HEPES 20 mM) supplemented with 10 mM glucose. After 24 h of treatment with 10% patients’ serum, HUVECs were incubated with 2.5 μg/mL Hoechst 33342, trihydrochloride, trihydrate  for 30 min, in the dark, at room temperature. Then, HUVECs were washed and incubated with 10 μM H2DCF-DA for another 15 min and then washed 3 more times and incubated without any fluorescent probes for another 15 min, in the dark, at room temperature. Immediately after this step, cells were imaged using a Nikon CSU-W1 Spinning Disk confocal microscope, using a 40x objective . The non-fluorescent dye H2DCF-DA is a chemically reduced form of fluorescein and is cell-permeable; once intracellular esterases cleave off the diacetate (DA) moiety, H2DCF becomes sensitive to oxidation by ROS: in its oxidized form, dichlorofluorescein (DCF) is highly fluorescent and easily detectable. Thus, cells were excited with a laser at wavelengths 405 nm and 488 nm for Hoechst and DCF, respectively. Light emission was detected using 455/50 and 520/40 filters for Hoechst and DCF, respectively. The same settings  were used for imaging in all experimental groups. In order to prevent DCF photodynamic reactions, fields of view search and focusing were performed using a Hoechst signal. Quantification of DCF fluorescence was performed using ImageJ software.ROS production was quantified using 2′-7′-dichlorofluorescin diacetate . Cells were excited with a laser at wavelengths 488 nm and 561 nm for MTG and MitoSOX, respectively. Light emission was detected using 520/40 and 605/52 filters for MTG and MitoSOX, respectively. The same settings  were used for imaging all experimental groups. In order to prevent MitoSOX photodynamic reaction, fields of view search and focusing were performed using a MTG signal. MitoSOX fluorescence intensity was quantified using NIS-Elements software (Nikon Corporation).Mitochondrial ROS production was quantified by MitoSOX™ Red  as previously described and validated ,143,144.11-BODIPY581/591 (ThermoFisher Scientific) as previously described [11-BODIPY581/591 for 30 minutes (37 °C in the dark) and then fluorometrically measured (with an excitation wavelength of 581 nm and an emission wavelength of 591 nm) using a microplate reader.MDA and 4-HNE levels were measured by using commercially available kits , following the manufacturer’s instructions . We quanescribed ,146. BriImmunoblotting assays were performed as previously described by our group ,147; thep < 0.05, was tested using the two-way ANOVA followed by Tukey–Kramer multiple comparison test or the non-parametric Mann–Whitney U test, as appropriate. Statistical analyses were performed using GraphPad 9 . Statistical significance, set at Taken together, our results indicate that the serum from COVID-19 patients who did not survive induces oxidative stress, lipid peroxidation, and ferroptosis in human endothelial cells through a mechanism that depends on TNF-α.


Layered double hydroxides (LDHs) constitute a unique group of 2D materials that can deliver exceptional catalytic, optical, and electronic performance. However, they usually suffer from low stability compared to their oxide counterparts. Using density functional calculations, we quantitatively demonstrate the crucial impact of the intercalants  on the stability of a series of common LDHs based on Mn, Fe, and Co. We found that intercalation with the singly charged lactate results in higher stability in all these LDH compounds, compared to neutral water and doubly charged carbonate. Furthermore, we show that the dispersion effect aids the stability of these LDH compounds. This investigation reveals that certain intercalants enhance LDH stability and alter the bandgap favourably. Carbonate [By having atomically thin layers and internal nano-space intercalation, layered materials are at the frontier of nanomaterial research ,2,3. Amo capture ,6, templ capture , photoca capture ,8, super capture , membran capture , and dru capture ,12. LDHsarbonate ,15,16,17arbonate , sulphatarbonate ,16, and arbonate  are somearbonate . Recent arbonate . Therefo2Al(AlSi)O2(OH)10·2.25H2O) [0.7Al0.3(OH)2(CO3)0.15·0.63H2O) [Carbonate, one example of an intercalating anion, has three oxygens and therefore possesses a strong hydrogen bonding capacity with the interlayer surface of an LDH, thus showing a high adsorption tendency with LDHs ,22. Prev1/3) LDH , strätli2.25H2O) , and hyd0.63H2O)  were expoduction . Most ofoduction . In partoduction ,27.Given that the monometallic LDHs warrant further investigations, in this work we theoretically quantify the critical role of intercalant anions on the stability of three transition metal (TM) based LDHs, i.e., Mn, Fe, and Co, in monometallic form. We systematically investigate the role of the neutral water molecule, a single negatively charged lactate radical, and a double negatively charged carbonate radical as intercalants in these LDHs. Moreover, we also study the effect of intercalant anions on the bandgap of LDHs, which determines their optical properties. The latter part can aid in designing LDH-based photocatalysts, electrocatalysts, and photovoltaics.k-point grid with 0.07 Å−1 spacing was used for integration over the Brillouin zone throughout all geometry optimisations. This spacing produced a −1 and energies smaller than 10−5 eV. When a supercell contained more than one magnetic ion, both ferromagnetic and antiferromagnetic spin alignments were examined, and the lowest energy configuration was used. Using true magnetic ground state is critical for obtaining realistic geometries and total energies [We performed spin-polarized first-principles density functional theory (DFT) calculations ,29 usingmisation , performenergies .U) term for the 3d electrons to correct the inherent and superficial electronic delocalisation associated with the LDA functional [U values has been demonstrated to offer excellent electronic structures at a reasonable computational cost [We used an ad hoc Hubbard  for all the intercalated compounds are shown in U formalism, we demonstrated that Mn-, Fe-, and Co-based layered double hydroxide compounds were stable against decomposition to the respective most stable oxides. Furthermore, in Mn- and Co-based LDH compounds, this stability is enhanced with either water, lactate, or carbonate intercalation. However, the most significant margin of stability was achieved for lactate intercalation. In Fe-based LDH compounds, water and carbonate intercalation reduced the margin of stability against decomposition to Using density functional calculations within the LDA +


Salt stress is a severe type of environmental stress. It adversely affects agricultural production worldwide. The overproduction of reactive oxygen species (ROS) is the most frequent phenomenon during salt stress. ROS are extremely reactive and, in high amounts, noxious, leading to destructive processes and causing cellular damage. However, at lower concentrations, ROS function as secondary messengers, playing a critical role as signaling molecules, ensuring regulation of growth and adjustment to multifactorial stresses. Plants contain several enzymatic and non-enzymatic antioxidants that can detoxify ROS. The production of ROS and their scavenging are important aspects of the plant’s normal response to adverse conditions. Recently, this field has attracted immense attention from plant scientists; however, ROS-induced signaling pathways during salt stress remain largely unknown. In this review, we will discuss the critical role of different antioxidants in salt stress tolerance. We also summarize the recent advances on the detrimental effects of ROS, on the antioxidant machinery scavenging ROS under salt stress, and on the crosstalk between ROS and other various signaling molecules, including nitric oxide, hydrogen sulfide, calcium, and phytohormones. Moreover, the utilization of “-omic” approaches to improve the ROS-regulating antioxidant system during the adaptation process to salt stress is also described. During their life cycle, plants encounter diverse biotic and abiotic stresses, varying from salinity, drought, cold, flooding, heat, mineral deficiency or excess, and exposure to toxins . Stress 1O2), superoxide radical (O2−•), hydroxyl radical (HO•), hydrogen peroxide (H2O2), alkoxyl radicals (RO•), and peroxy radicals (ROO•) (ROS include singlet oxygen (s (ROO•) . They ars (ROO•)  15,16].,16.1O2),The cell wall, plasma membrane (PM), peroxisomes, mitochondria, and chloroplasts are the primary sites of ROS generation in plant cells  21,22].,22.21,223Chl*), inhibiting energy transmission to other molecules. If 3Chl* cannot be effectively quenched, it reacts with triplet oxygen, generating 1O2 [1P680*), transfers an electron to pheophytin, and subsequently to a quinone in the PSII reaction center. In adverse conditions, if this quinone acceptor is not capable of taking any electrons, pheophytin couples with P680, forming the triplet photoexcited state 3P680* [3P680*; however, the distance between β-carotene and 3P680* is too great, and the quenching is unsuccessful, leading to the production of 1O2 [1P680* in PS II and enhances the production of 1O2 [1O2. However, photoreduction of the molecular oxygen at PSI results in the production of O2−• (Mehler reaction), which is further converted into H2O2 via superoxide dismutases (SODs) [2+ transform H2O2 and O2−• into HO• [2−•, which yields H2O2 via copper-zinc-SOD (CuZn-SOD) and manganese-SOD (Mn-SOD) [2−•, the latter being converted to H2O2 through urate oxidase and SOD [Carotenoids such as lutein and zeaxanthin extinguish triplet-excited Chl (ting 1O2 . Furthere 3P680* . Theoretn of 1O2 . Addition of 1O2 . Converss (SODs) ,31. Theninto HO• . In the (Mn-SOD) ,31. In p(Mn-SOD) . Further and SOD ,34,35.2O2 is produced in the peroxisomes by flavin oxidase and from disproportionation of O2−• [2O2 in the peroxisome [2O2 via the ascorbate-glutathione (AsA-GSH) cycle and produces AsA in the peroxisomes [2−• in the endoplasmic reticulum. β-oxidation, urate oxidase (UO), and glycolate oxidase generate H2O2 and O2−• in the glyoxysomes [Apart from β-oxidation, H of O2−• ,36. Furtroxisome . MDHAR qoxisomes ,39. NADPoxisomes ,40. Moreoxisomes ,41, whiloxisomes . Additiooxysomes . Finallyoxysomes .In plant cells, physiological ROS function as secondary messengers, being redox-signaling molecules in various signaling pathways that regulate the response to adverse environmental conditions ,45. In cThe antioxidant enzymes—including CAT, SOD, MDHAR, APX, guaiacol peroxidase (GPX), DHAR, and GR—localized in diverse plant cell compartments comprise the plant antioxidant defense system A. In the2−• into O2 and H2O2, ensuring its elimination [2−• into H2O2 and O2 as follows:2•−+ SOD − M2+ → O2 + M+--------Step 1O2•−+ 2H+-SOD − M → H2O2 + M2+-------Step 2OSOD (E.C.1.15.1.1) is a metalloenzyme present in all aerobic organisms. It represents the first bulwark against ROS-induced damage owing to different environmental stresses. SOD catalyzes the conversion of Omination . Thus, tmination . Based omination . SOD iso2•−+ O2•−+ 2H+ → 2H2O2 + O2OThe overall reaction is presented as the following equation:Superoxide free radicals are produced at any location of the ETC; hence, SOD activation is evident in mitochondria, peroxisomes, and chloroplasts . The Hab2O2 and molecular oxygen are both byproducts of the enzymatic reaction catalyzed by SOD. H2O2 regulates different signaling activities. It is an essential signal during plant-microbe interactions, the response to inflammation, the closing of the stomata, and osmotic and excessive light stress [The abundance of different SOD isoenzymes in plants is influenced by climatic conditions and the growth phase of the plant . Compount stress . Abiotict stress .2 [2O2 into water and O2. The iron prosthetic group of the enzyme is essential for its catalytic activity. This enzyme shows higher reduction activity toward H2O2 than toward organic peroxides (R-O-O-R). CAT eliminates H2O2 with minimal energy use [2O2 [CAT (E.C.1.11.1.6) is a tetrameric iron-containing redox enzyme present in all organisms exposed to O2 . This energy use  and a veergy use . β-oxidause [2O2 . Based ouse [2O2 . Most ofuse [2O2 . However2O2 produced during photorespiration and is widely present in pollen and seeds [2O2 produced by the β-oxidation of lipids in the glyoxysomes [2O2 [CAT1 is critical for the elimination of Hnd seeds , but lesoxysomes . Stressfmes [2O2 .2O2 to water and O2 or other oxidized compounds such as lower alcohols , aldehydes , etc. In the latter, they convert lower amounts of H2O2 by oxidizing various hydrogen donors such as small-molecule alcohols, aldehydes, and ascorbic acid (AA) as follows [2 + H2O2 → R + 2H2ORH2H5OH → CAT-Fe-OH + H2O + CH3CHO (peroxidatic reaction)CAT-Fe-OOH + C2O2 → CAT-Fe-OH + H2O + O2 CAT-Fe-OOH + HCAT isoforms act in two modes: catalytic and peroxidatic. In the former, they catalyze the direct conversion of H follows :RH2 + H22O2. Depending on the reaction mode, catalases may be further classified asThus, CATs generate useful compounds and water from harmful compounds and HHPI catalase: comprises isoenzymes with dual function, participating in both catalytic and peroxidative reactions.HPII catalase: comprises isoenzymes that only participate in peroxidative reactions; these CATs play a crucial role in protecting plants from abiotic stress caused by different factors . Plants 2O2 pretreatment enhanced CAT activity and prevented the deleterious effects of salinity. Furthermore, a higher CAT activity was essential for the existence of plants under reasonable metal stress. However, Spartan metal stress causes the permanent impairment of CAT. The function of CAT is taken over by other enzymes, which ensure abiotic stress is managed [Gondim et al.  reported managed .2O2 through the AsA-GSH pathway  [2O2 in the cytosol and chloroplasts, while CAT primarily acts in the peroxisomes. APX, in the presence of a reducing agent such as AA, reduces H2O2 to water while the other reactant is oxidized . This enzyme is very unstable in the absence of AA. At concentrations AA < 20 µM, the activity of APX drastically decreases [2O2 scavenger than CAT because it is more extensively distributed and has a stronger affinity for H2O2. APX acts as an antioxidant by the following reaction:2O2 + AA→2H2O + DHAHAPX (E.C.1.1.11.1) is a key enzyme found abundantly in plants and algae . It is dpathway) . APX mosecreases . Under s2O2 [2O2 levels increase and cause aberrant stomatal closure [2O2-mediated-enhanced activation of heat shock proteins [To provide significant protection during oxidative stress, the enzymatic and non-enzymatic antioxidant molecules closely coordinate their actions. By keeping the redox equilibrium under stress, APX connects the two pathways. Furthermore, it has a particular affinity for H2O2 . Plants  closure . In APX1proteins ,74. Furtproteins . Moreoveproteins .2O2 and concomitantly oxidizes AA [MDHAR (E.C.1.6.5.4) restores AA from MDHA in the presence of NADPH as a reducing agent . Subsequdizes AA . MDHAR hdizes AA .+MDHA + NADPH→AA + NADPThe monovalent oxidation of AA results in MDHA. In the absence of MDHAR, MDHA will convert non-enzymatically to AA and DHA. Then, DHA is converted to AA by DHAR as GSH oxidizes . A rapidDHAR (M.C.1.8.5.1) is an enzyme that catalyzes the reduction of DHA to AA in the presence of GSH as an electron contributor . TogetheDHA + 2GSH→AA + GSSGThe catalytic mechanism of DHAR involves three steps . In the 1O2 [GR (E.C.1.6.4.2) belongs to the oxidoreductase class of enzymes and needs NADPH as a reducing agent to convert GSSG to GSH. The isoforms of GR are abundant in chloroplasts, cytosol, and mitochondria . In vari1O2 .+GSSG + NADPH → 2GSH + NADP2O2 and DHA to H2O and AA [GR exists mostly as a homodimer with a molecular weight ranging from 100 to 150 kDa. It comprises a single FAD molecule per monomer. In the absence of thiols, GR tends to aggregate into tetramers and even larger forms . The catO and AA . An enhaO and AA . The GR O and AA .2O2 generated by regular metabolism and by exposure to different stress conditions. Hence, it is also called the “stress enzyme” [2+ and four disulfide bridges. It is dynamically involved in the biological production of lignin and contributes to the defense against biotic stress by breaking the indole acetic acid (IAA) and consuming the H2O2 in the process [2O2 and lipid peroxides to water and fatty alcohols, using GSH as the cofactor. Subsequently, GSH is oxidized to GSSG. The enzyme GPX possesses cysteine in the active location instead of selenocysteine; thus, GPX may present selenium-dependent and -independent forms [GPX (E.C.1.11.1.7) is composed of 40–50 kDa molecular weight monomers. It actively decomposes H enzyme” . It cont process ,100. Pla process . This ennt forms .2O2 as it functions both outside and inside the cell, in the organelles (the cytosol and vacuole).The GPX enzyme is considered the primary enzyme in the elimination of H2O2 + GSH → H2O + GSSGHGPX is engaged in different biosynthetic pathways, such as the synthesis of lignin, which contributes to cell wall strengthening, the catalytic breakdown of IAA, and the production of ethylene (ET). It contributes to the rehabilitation of the biotic and abiotic stress-induced lesions. In plant cells, its activity is enhanced by exposure of the plant to different stress factors . Around The non-enzymatic antioxidants, which include AA, GSH, alpha-tocopherol, carotenoids, different phenolics, flavonoids, and the amino acids and osmolyte proline, make up the other half of the antioxidant machinery in plants. They play a crucial role in plant development and growth by adjusting cellular processes including cell division, cell expansion, maturity, and death. Moreover, they serve to protect various cell components from injury .AA is the most prevalent, water-soluble, and extensively researched antioxidant agent . Approxi2O2, HO•, and O2−• and restores α-tocopherol from tocopheroxyl free radicals produced through metabolism or induced by various types of stress. Thus, it contributes to the protection of cell membranes and the regeneration of the lipoproteins in their structure [MDAR increased the tolerance of tobacco plants to ozone, high salinity, and polyethylene glycol. This was attributable to the subsequent increase in AA levels [The highest concentration of AA is found in the leaves. Furthermore, a linear relationship exists between the amounts of chlorophyll and AA. AA represents one of the most effective antioxidant agents; it counteracts ROS by donating electrons in different enzymatic and non-enzymatic pathways. AA directly scavenges Htructure . AA alsotructure . The firtructure . Additiotructure . AmplifiA levels . FurtherA levels .Nearly all subcellular compartments, including the mitochondria, chloroplasts, vacuoles, endoplasmic reticulum, peroxisomes, cytosol, and apoplasts, contain high amounts of glutathione, a hydrosoluble antioxidant thiol tripeptide (γ -glutamyl-cysteinyl-glycine) . Among t1O2, O2−•, HO•, and H2O2, protecting diverse biomolecules. Subsequently, glutathione-derived derivatives, or GSSG, are formed as byproducts. GSH also plays an important role in reproducing AA to yield GSSG using NADPH as the reducing power [GSH is one of the most important antioxidants, as it helps maintain a low level of ROS . It actsng power . The celng power . ROS-indng power . Thus, tVTE4 [Among the four existing isomers , α-tocopherol has the strongest antioxidant properties. Tocopherols are exclusively produced by photosynthetic organisms and are only found in the fresh tissues of plants . α-tocopVTE4 . TocopheVTE4 ,126. TheVTE4 . By inteVTE4 . AdditioVTE4 . At the VTE4 . Yu and VTE4  found thVTE4 . SubsequVTE4 . TocopheVTE4 .1O2 without deterioration. The chemical reactivity of β-carotene with free radicals, such as O2−•, HO•, and ROO•, arises from the conjugated double-bonded structure, which allows the delocalization of unpaired electrons and is the primary cause of its antioxidant capacity [1O2, which produces heat as a by-product; (c) they inhibit the formation of 1O2 by counteracting 3Chl* and Chl*; and (d) they release energy when it exceeds the required amount through the xanthophyll cycle [Carotenoids, a class of phytonutrients generated by plants, algae, certain bacteria, and fungi, are pigments also known as tetraterpenoids . They arcapacity . Higher capacity . Carotenll cycle .2O2, HO•, and O2−•. Furthermore, anthocyanidin acts as a strong inhibitor of fatty acid oxidation [1O2, reducing its damaging consequences on the external layers of the chloroplasts. The concentration of flavonoids in plant cells typically exceeds 1 mM. Various flavonoids act as possible regulators of lipoxygenase, the enzyme that converts polyunsaturated fatty acids into corresponding peroxides and hydroperoxides [2O2 and good inhibitors of lipid oxidation. This is one of the most actively researched features of flavonoids.Flavonoids are mostly distributed throughout the plant world and are particularly abundant in the leaves, floral organs, and pollen . Based oxidation . The abixidation . Due to eroxides . Owing t1O2. In plants, it builds up significantly under stress, either as a result of enhanced production or decreased breakdown [Proline is a potential non-enzymatic antioxidant that counteracts the negative consequences of various ROS in the components of different plant species . Prolinereakdown . Plants reakdown  reportedreakdown . ProlineRedox reactions are ubiquitous in all living organisms and regulate the generation of ROS in plant cells . The exi2 and inhibit its fixation, resulting in a decrease in molecular oxygen and an overgeneration of ROS. The activity of the chloroplasts is impaired, thereby hindering the process of photosynthesis [2O2 and a 2- to 3-fold increase in the levels of thiobarbituric acid. Furthermore, the impact of the salinity stress on the generation of ROS differed between different developmental stages and tissues. For example, the root is more sensitive to salinity-induced oxidative stress than young and mature leaves. Cheng et al. [2O2 and O2−• , electrolyte leakage (158%), and malondialdehyde . Compared with control, salinity stress induced a 2-fold increase in electrolyte leakage and MDA levels in sweet peppers and in the levels of electrolyte leakage, MDA, H2O2, and O2−• in mung bean (100 mM NaCl) [2O2 (50%) [2O2 increased by 78% in FH-572, whereas it reduced by 20% in FH-621, indicating that FH-621—not FH-572—is a salt-tolerant genotype [2O2 and MDA) during high salinity stress in Medicago truncatula. Ailanthus altissima exposed to 150 mM NaCl had a higher level of antioxidant activity than non-stressed Ailanthus altissima. However, the level of H2O2 did not differ significantly between the two groups, indicating a connection between the antioxidant defense machinery and the enhanced insensitivity of plants in harsh environmental conditions [Exposure of plants to several abiotic stresses under field conditions resulted in the generation of oxidative stress via the overproduction of ROS. ROS are produced mainly in chloroplasts and, to a lesser extent, in PMs, mitochondria, apoplasts, and peroxisomes ,23. Mostynthesis . Howeverynthesis . Rehman ynthesis  reportedg et al.  found thg et al.  showed tmM NaCl) . FurtherO2 (50%) . The deggenotype . Similargenotype . Moreovegenotype  revealednditions . CollectIn adverse environmental conditions, plants activate their antioxidant defense machinery to minimize the damage induced by oxidative stress. The antioxidant defense response differs depending on the species, genotype, stress type, and duration of exposure to stress. Various plant species have different methods to increase their antioxidant capacity. Regulation of the antioxidant defense system counteracts the effects of high salinity stress and increases stress tolerance in various plant species. The response and the potency of the antioxidant activity vary depending on the developmental stage, type of tissue, salinity degree, and duration of exposure to stress .For example, Vighi et al.  reportedVicia faba, when the levels of H2O2 exceeded 90%, the expression of CAT, SOD, and GR at the mRNA level and that of AsA at the protein level were elevated, demonstrating the regulation of the antioxidant response by high salinity and its mitigation [OsSOD3-Cu/Zn, OsGR2, OsGR3, and OsAPX3 are the distinct markers of the salt-tolerant rice cultivar, not the salt-sensitive one. Furthermore, the salt-tolerant wheat cultivar Suntop presented higher activities of the antioxidant enzymes CAT, APX, SOD, POD, and GR than the salt-tolerant wheat cultivar Sunmate. Additionally, the barley salt-tolerant cultivar, CM72, presented higher activities of the antioxidants than the sensitive cultivar. These results indicate that an elevated level of antioxidants or enhanced antioxidant activity contributes to salt tolerance in plants . Anothertigation .2O2 level in salt-stressed wheat seedlings (compared with not-stressed wheat seedlings). Similarly, combinatorial treatment with humic acid and jasmonic acid (JA) also enhanced APX activity in sorghum, improving its salt tolerance [2O2 levels (2.5-fold), and increased O2−• levels (1.7-fold) [Phaseolus vulgaris and Triticum aestivum, Glycyrrhiza glabra root extract or 6% Moringa oleifera leaf extract ameliorated the salinity stress by enhancing the antioxidant activity [Sesamum indicum [The role of the antioxidant “defense system” in improving salinity tolerance was demonstrated by using natural or chemical protective compounds to regulate the antioxidant activity. For instance, Alsahli et al.  reportedolerance . In saliolerance . Further.7-fold) . Exogeno.7-fold) . In soyb.7-fold) . Similar.7-fold) . Santand.7-fold)  showed tactivity ,177. Las indicum . These rCucumis melo and Solanum tuberosum have a higher level of proline than salt-sensitive cultivars [Solanum lycopersicum exhibited a higher accumulation of proline than non-stressed Solanum lycopersicum [Calendula sp. and Vicia faba [Exogenous application of proline enhanced stress tolerance  and planultivars . Similarpersicum . Additiocia faba ,184. Concia faba ,186. Usucia faba .Arabidopsis showed that P5CS1 regulates proline synthesis, with its expression being highly elevated under high salinity stress [P5CS1 lead to a severe decrease in salinity stress-induced proline expression. Subsequently, growth is impaired, and the levels of ROS increase, indicating that these mutants are hypersensitive to high salinity stress [The biosynthesis of proline through the glutamate pathway uses two molecules of NADPH for one molecule of proline, removing the electrons from the chloroplast and buffering the redox potential of the cell . Proliney stress . Knockouy stress . Glyciney stress . The effy stress ,192. Higy stress . These r2O2, and other ROS, H2S, calcium (Ca2+), and phytohormones crosstalk and control diverse biological processes and the expression of genes implicated in salt stress tolerance [Salinity tolerance is regulated by an intricate gene network and affects diverse cellular processes in plants. Different signaling molecules such as NO, Holerance ,10,11. T2O2 formation results in oxidative stress, and its accumulation causes cellular and molecular damage, activating programmed cell death [2O2 serves as a signal to regulate various biological processes and the crosstalk among different signaling pathways [2O2 and NO signaling pathways are strongly correlated with the plant response to various environmental stimuli [2O2 and NO. Zhao et al. [AtNOA1 in Arabidopsis, resulting in a subsequent reduction in NO levels. Conversely, high salinity stress moderately elevated the transcript-level expression of OsNOA1, a homolog of AtNOA1 in rice [2O2 enhanced salt tolerance [2O2 also regulates translation and post-translational changes. In Bermuda grass, NO and H2O2 stimulated the activities of CAT and SOD via ABA [2O2 by regulating the expression of SOD. Thus, SA and H2O2 work together in a self-amplifying manner. H2O2 at higher concentrations causes nitrosative or oxidative stress, whereas at basal levels it functions as a signaling molecule, regulating plant response under diverse stress conditions.Hll death . Its genpathways . H2O2 an stimuli . High sao et al.  found th in rice . Furtherolerance . Besides via ABA , affecti via ABA . SubsequArabidopsis callus exposed to salt stress (100 mM NaCl) triggered the accumulation of NO and a subsequent increase in the expression of H+-ATPase in the PM [+/K+), resulting in enhanced susceptibility to salt stress [2O2 accumulation, which increases the H+-ATPase influx into the PM. The combined effect of NO and SA enhances the absorption of Mg2+/Ca2+ and reduces the uptake of Na+ in conditions of salt stress [NO is a key gasotransmitter that regulates various signaling pathways and a range of physiological processes during salt stress in plants . The cron the PM . Howevert stress . The supt stress . SA intet stress .2S help counteract the effects of salt stress in plants. In cucumber, NO application under saline stress modulates proline metabolism, increasing the accumulation of free proline ratio; thus, it maintains the turgor potential and protects cucumber seedlings [2 improves the salt tolerance of mustard by enhancing the antioxidant activity and the accumulation of proline and glycine betaine. Subsequently, a decline in thiobarbituric acid-reactive substances, electrolyte leakage, and H2O2 is observed [Lactuca sativa, NaCl application activates ionic, osmotic, and oxidative stress, leading to hormonal imbalances and reduced plant growth. Supplementation of NO results in decreased Na+ accumulation and the maintenance of mineral balance, thereby improving the photosynthetic rate and growth [Crocus sativus treated with NO displays vigorous growth during high salinity. NO application induced the biosynthesis of secondary metabolites, enhanced the deposition of compatible solutes, and enhanced the activities of antioxidant enzymes, while exogenous application of SA did not ameliorate the growth of plants under salt stress [Brassica napus L. seedling roots, concomitant application of NO-releasing substances and of melatonin neutralized the inhibitory effect of NaCl on seedling growth and reestablished redox and ion homeostasis, which was demonstrated by a decrease in the overproduction of ROS, the ratio of Na+/K+, and the synthesis of thiobarbituric acid-reactive substances. Melatonin inhibited the effects of salinity stress via a NO-dependent mechanism [Sulfur is a primary biocomponent and is present in methionine, GSH, cysteine (Cys), coenzyme A, thioredoxin, sulfo-lipids, and iron–sulfur (Fe–S), which control cellular processes in high salinity conditions ,211. Addeedlings . The exoobserved . In Lactd growth . NO signt stress . In saltechanism . Thus, N2+ and ROS are well-known intracellular signals. Under severe salt stress conditions, the concentration of cytosolic Ca2+ is significantly elevated, activating specific signaling pathways [+ and Na+, accumulation of compatible solutes, and an increase in the expression of stress-related genes [Salt overly sensitive (SOS) pathway plays a crucial role in the response to salt stress. It regulates the membrane conductance in the epidermal cells of the root when exposed to high salinity. Thus, it helps detoxification by the epidermal cells of the root by removing the ions from the root . The SOSpathways . Salt sted genes .2. Consequently, reduction of carbon and utilization of photosynthetic NADPH in the Calvin cycle decrease, leading to electron holes in PS I and electrolyte leakage [2S and GSH, which crosstalk with NO [2O2 in the root tissue and young leaves, which killed the plant [+ than non-supplemented plants. Furthermore, leaves did not display any symptoms of salt injury, and the integrity of the membrane was maintained. Owing to the above-mentioned roles of abiotic-stress-induced intracellular ROS, a question was raised on whether intracellular amounts of NO and ROS can contribute to the protection of the distressed leaf protoplasts. ROS also activate Ca2+-permeable channels such as respiratory burst oxidase homolog (Rboh), which is assumed to represent a positive feedback loop that induces Ca2+ and ROS signals in root cells. In Arabidopsis, RHD2/RbohC influenced the generation of ROS, regulating root hair elongation in a Ca2+-dependent manner [The production of ROS in plant organelles is unavoidable as they are byproducts of metabolic processes ,19,31. H leakage . Glycola leakage . The que leakage . Other s with NO ,212. Phy with NO . The neghe plant . Szepesihe plant  reportedt manner .2+ levels via depolarization and activation of hyperpolarization-activated Ca2+ permeable channels in the PM. This increase plays an important role in ROS signaling and salt stress tolerance [2+ pathway in plants [2+/Calmodulin-dependent Protein Kinases (CDPKs) were more evident in halophytes compared with glycophytes under salt stress. The calcium-binding proteins play an important role as amplifiers in early calcium influx during salinity stress. Ca2+ signaling has a significant impact on signaling mechanisms in salt stress. Two-pore channel 1 is involved in the dissemination of the signal (Ca2+) in salt stress and the activation of the plant defense [Salt stress increases cytosolic Caolerance . Besidesn plants . This con plants  demonstr defense . Further defense . The pro defense ,212,215.2S serves as a signaling molecule and plays an important role in seed germination, postharvest senescence, and adventitious rooting. Furthermore, it also has a protective role in multifactorial stress conditions, including biotic and abiotic stresses [2S donors such as phosphinodithioate, morpholin-4-ium 4 methoxyphenyl, NOSH-aspirin, CaS2, AP39, sodium hydrosulfide, diallyl trisulfide, and dialkyldithiophosphate, have been identified and characterized. NOSH-aspirin simultaneously released two gasotransmitters, for instance, NO and H2S [2S, as this enhanced the activity of the antioxidant enzymes CAT and SOD [2S through post-translational changes. By regulating the antioxidant defense system, H2S also preserves membrane integrity and ROS homeostasis, increasing the tolerance level under saline conditions. Mostofa et al. [2S contributes to the maintenance of the Na+/K+ ratio during salt stress. In rice, salt stress increases Na+ concentration and decreases K+ concentration, resulting in an increased Na+/K+ ratio in the root and leaves. Exogenous application of H2S maintained K+/Na+ homeostasis during salt stress. Furthermore, under high salinity stress, H2S acts synergistically with NO in response to salt stress [2S revealed that in stressful conditions, NO induces H2S accumulation in plants and increases stress tolerance; H2S further functions as a downstream signal. Nitrosothiol is a newly identified signal molecule generated when H2S responds to NO Cys and plant hormones and contributes to NO-mediated salt stress tolerance.Hstresses ,174. It stresses . Further and H2S . Further and SOD . Hence, a et al.  demonstrt stress . The dow2+ also mediates the production of special metabolites induced in response to SA, ABA, and JA [2+ and MT—and how they influence the development of phenolic compounds under salt stress—were examined in Dracaena kotschyi. Exogenous application of NaCl decreases the dry biomass of shoots; however, it enhances H2O2 production, electrolyte leakage, the scavenging capacity of 2,2-diphenyl-1-picrylhydrazyl, and the expression of TAL, PAL, and RAS . Pretreatment of D. kotschyi with an inhibitor of the melatonin pathway had no influence on the Ca2+-induced generation of phenolic compounds in salt-treated plants [D. kotschyi with a PM channel blocker during salt stress had the opposite effect. These results indicate that melatonin induces the biosynthesis of phenolic compounds only when there is an influx of Ca2+.Calcium is one of the key signaling molecules and secondary messengers that crosstalks with other signaling molecules to minimize the impact of salt stress. The activities of phenylalanine ammonia lyase (PAL) and flavonoids were increased after the incorporation of calcium chloride or an ionophore into the culture medium . Further, and JA ,170,222.d plants . However2O2 work together with Ca2+ to form an intricate regulatory network to mitigate the effect of salinity stress [2O2, and sodium nitroprusside exhibits a positive linear relationship with the germination rate and germination index, while under salt stress, a negative linear relationship was observed with the mean germination time. Pretreatment of seeds enhanced their germination and the quick establishment of the seedling in the saline soil. Additionally, pretreatment with NaCl reduced α- and β-amylase activities. This inhibited starch hydrolysis, impairing seed germination. The above-mentioned pretreatment reduced the negative effect on these enzymes. Another study demonstrated that external application of calcium chloride and H2O2 might decrease the harmful effect of multifactorial abiotic stress on the activity of amylase and rescue seed germination [2+, NO, and H2O2 induced the activity of amylase. The increased starch degradation enhanced seed germination and sprouting in quinoa, minimizing the negative impact of salt stress.NO and Hy stress ,230,233.mination . InteresADC was elevated under salt stress conditions  [2O2, and ABA [Lactuca sativa [2+. They regulate ion channel activity by modulating their conductivity under salt-stress conditions [2+ might have a detrimental effect on the standard cellular metabolism in stressful conditions. Polyamines trigger the efflux of Ca2+ by activating the membrane Ca2+-ATPase and the PM channel. Thus, they play an important role in maintaining intracellular Ca2+ homeostasis and sustaining a steady plasmatic level [2+ signaling is poorly understood.Polyamines such as putrescine, spermine, and spermidine have been implicated in a wide range of plant developmental processes. They also regulate the adaptive response to various stress conditions . Additioditions) . The met and ABA ,235,237.a sativa . Additionditions . Moreoveic level . However2+. Furthermore, it may influence calcium ion channels, eventually activating Ca2+ signaling. The mechanisms underlying this crosstalk are largely unknown. The activation of NO synthase, which catalyzes NO synthesis, involves activation of calmodulin and Ca2+ signaling [2+ during salt stress recovery [2+ signaling plays an essential role in the response and adaptation of plants under salt stress.Numerous studies have demonstrated the tight regulation of NO metabolism under salt stress ,216. In ignaling ,206. Morrecovery . Therefo2O2, NO, and H2S. For instance, supplementation of NO in salinity-stressed lettuce reduces the accumulation of Na+ and regulates the concentrations of mineral nutrients, thereby maintaining the photosynthetic rate and restoring vegetative growth. Furthermore, NO also restored the osmotic balance, stimulated the antioxidant system, and reinstalled the equilibrium of plant hormones, improving the plant’s tolerance to high salinity stress [+/Na+ ratio. Exogenous application of 24-epibrassinolide, sodium nitroprusside (SNP), or their combination enhanced endogenous ABA levels in Brassica juncea via nitrogen and proline metabolism [+/Na+ ratio. By generating ROS such as hydroxyl radicals (HO•), superoxide (O2−•), and H2O2, it also modulates the cellular redox pathways. The formed free radicals negatively impact cellular processes by inducing lipid peroxidation and damaging nucleic acids and proteins. Therefore, they impair fundamental processes such as plant growth and development, gaseous exchange, nitrogen, carbon, and proline metabolism [Amaranthus caudatus [Arabidopsis, GA and ET positively regulate hypocotyl elongation. GA alone is inefficient; however, it acts synergistically with ET and enhances the growth of emerging roots. These results suggest that GA and ET do not have additive results but rather act synergistically.In agriculture worldwide, the quality and yield of crops are significantly affected by high salinity stress. The capacity of plants to tolerate this type of stress can be enhanced by the exogenous application of Hy stress . High satabolism . High satabolism ,242. Thetabolism . Phytohotabolism ,5,244. Gtabolism . Furthercaudatus . Furthercaudatus . In ArabArabidopsis, the exogenous application of sodium nitroprusside and ACC counteracts the negative impact of salt stress on seed germination [ACS2 expression was elevated by sodium nitroprusside. These findings demonstrate the significance of NO and ET during salt stress. Supplementation of ACC enhances seed germination under oxidative stress; seed germination is activated by H2O2. Nevertheless, NO-treated Arabidopsis was not substantially affected, while the seeds of the ET-insensitive mutant were not affected in any way.In plants under salt stress, seed germination is influenced by NO and RT. In mination . ConversMatricaria chamomilla, external application of SA (500 μM/L) markedly enhanced plant growth under salt stress and normal conditions [Arabidopsis. These findings demonstrate that the beneficial effect of SA on salinity stress might depend on the concentration of salt and may vary between different plant species. The detailed mechanism underlying the effect of SA on plants exposed to salt stress remains unknown.SA plays a crucial role in various physiological and biochemical processes and has significant effects on the tolerance to biotic and abiotic stress ,250,251.nditions . Howevernditions  and Arfanditions  showed tnditions  reportedOsIAAs respond to different abiotic stresses, suggesting a link between abiotic stresses and plant growth. Additionally, the expression of OsIAA18 in rice was induced by phytohormones and various abiotic stresses, including high salinity, drought, and treatment with abscisic and IAA treatments [OsIAA18 enhanced the tolerance to salinity and osmotic stress in Arabidopsis [OsIAA6/OsIAA20 improves the tolerance to salinity and drought in rice [VvIAA18 increases the tolerance to salinity in tobacco [Auxin regulates diverse physiological and developmental processes in plants ,258,259.eatments . Overexpbidopsis . Overexp in rice ,266, whi tobacco . TherefoIn the past few years, biotechnology approaches have been extensively used to develop cultivars with better nutritional quality, yield, and stress tolerance. Omics are the most prominent and efficient approaches to improving plant tolerance against various biotic and abiotic stresses. Plant biologists can utilize the omic tools, including genomics, transcriptomics, proteomics, metabolomics, phenomics, and ionomics, to identify genes, RNAs, proteins, metabolites, phenotypes, and ions belonging to the intricate regulatory mechanisms activated during the response to internal and external stimuli and during various developmental stages of plants .Chrysanthemum overexpressing DgNAC1 exhibited improved salt tolerance by reducing the content of O2−•, H2O2, and MDA and increasing the activities of CAT, POD, and SOD [Furthermore, deciphering the underlying mechanisms by which plants perceive and transduce signals that initiate adaptive responses is a prerequisite for the identification of key genes and pathways to cope with future challenges. Furthermore, decoding and understanding such a regulatory network of genes would help gain insight into plant biology and could be practically applied to improve the quality, productivity, resistance to disease, and stress resistance of crops. This will allow us to feed the ever-increasing human population. Hence, plants can be genetically engineered to increase their tolerance to oxidative stress and improve the response and activities of the antioxidant defense system. An overview of genetically engineered plants with increased activities of the antioxidant enzymes under salt stress is presented in  and SOD .PaSOD (Potentilla atrosanguinea) and RaAPX  displayed increased activity of APX and SOD. APX and SOD might be functioning as positive regulators, enhancing salt tolerance by the regulation of ROS signaling and by the stimulation of lignin biosynthesis [OsMYB6 exhibited enhanced tolerance to salt and drought stress compared with control rice. Under salt and drought stress, transgenic plants showed higher proline levels, higher SOD and CAT activities, and lower MDA and REL levels than wild-type plants [StCYS1 had a significantly higher salt tolerance than wild-type potatoes. Transgenic potatoes presented higher levels of proline, better scavenging of H2O2, and a superior integrity of the cell membrane than non-transgenic plants [OsSTAP1 presented enhanced salt stress tolerance, demonstrating higher CAT, POD, and SOD activities and lower Na+/K+ ratios in the shoots than wild-type rice [DnWRKY11 in tobacco; transgenic tobacco displayed longer root length, a higher germination rate, higher CAT, SOD, and POD activities, and lower MDA concentrations than wild-type tobacco. Soybean plants overexpressing GmMYB84 displayed a higher germination rate, longer root elongation, higher antioxidant enzyme activities, and higher proline content compared with wild-type plants [Similarly, transgenic potatoes overexpressing ynthesis . Rice ove plants . Potatoec plants . Rice ovype rice . Xu et aype rice  overexpre plants .OsEXPA7 significantly increased the salt tolerance of rice. Transgenic rice presented reduced levels of Na+ and increased levels of K+ in the leaves and roots of the plant. Compared with wild-type rice, transgenic rice presented higher activity of antioxidant enzymes (POD and SOD), lower accumulation of ROS and MDA, and higher proline accumulation. These findings indicate that OsEXPA7 plays a critical role in enhancing the tolerance to salt stress by regulating the transport of Na+ and scavenging ROS [MsWRKY11 showed a better tolerance to salt stress and longer hypocotyls than non-transgenic plants. Additionally, transgenic plants had higher contents of soluble sugar, chlorophyll, proline, CAT, and SOD, lower contents of O2−•, H2O2, and MDA, and lower relative electrical conductivity [NbExo70D1 had lower accumulation of ROS than control. Pretreatment with diphenylene iodonium, an inhibitor of NADPH oxidase, caused a reduction in salt stress-induced ROS production in the roots of both transgenic and wild-type plants. However, the activity of NADPH oxidase was lower in the transgenic plants than in wild-type plants under salt stress, suggesting NbExo70D1 participates in NADPH oxidase-mediated production of ROS [VvIAA18 in tobacco and E. coli; salt stress tolerance significantly improved in transgenic tobacco and E. coli. Quantitative reverse transcription PCR revealed that overexpression of VvIAA18 induced the expression of salt stress-responsive genes, such as LEA5, P5CS, POD, and SOD, under salt stress. In addition, transgenic tobacco had higher activities of POD and SOD and higher levels of proline and lower levels of MDA and H2O2 than wild-type tobacco. Zhu et al. [VvWRKY30 in Arabidopsis. This significantly improved the salt stress tolerance at different developmental stages, increased antioxidant activity, and decreased the ROS levels. Additionally, the concentrations of soluble sugar and proline and the transcript-level expression of genes involved in antioxidant biosynthesis and proline synthesis were also higher in salt-stressed transgenic plants than in wild-type plants. Overexpression of SsMAX2 in Arabidopsis resulted in decreased degradation of chlorophyll and enhanced accumulation of proline and soluble sugar. The antioxidant activities of SOD, APX, and POD were higher in transgenic plants than in wild-type plants, resulting in a significant decrease in H2O2 levels [MfWRKY70 in Arabidopsis, significantly improving the tolerance to salt, osmotic, and drought stress, enhancing the root growth, increasing the antioxidant activities of CAT, SOD, and POD, and maintaining ROS homeostasis. Tobacco overexpressing PeHSF displayed better control of the ROS homeostasis under salt stress than wild-type tobacco. Transgenic plants showed increased activities of APX, GPX, and GSH. These results indicate that PeHSF plays an essential role in the detoxification of ROS and in the transactivation of antioxidant genes under high salinity stress [ThHSFA1 exhibited increased tolerance to salt stress, reduced ROS levels, and enhanced antioxidant enzyme activities under salt stress [SlGGP-LIKE presented higher levels of reduced AsA, the xanthophyll cycle, and the AsA/DHA ratio in normal conditions, and higher levels of reduced AsA, reduced GSH, total GSH, higher AsA/DHA and GSH/GSSG ratios, and the xanthophyll cycle under salt stress conditions. Thus, transgenic plants showed better salt stress tolerance, higher photosynthetic, antioxidant, and D1 protein activities, and lower levels of ROS and membrane damage than wild-type plants. Overexpression of SlGGP-LIKE increases tolerance to salt stress by inducing the synthesis of AsA in tomatoes [Overexpression of ging ROS . Under huctivity . Tobaccon of ROS . Furthern of ROS  overexpru et al.  overexpr2 levels . Xiang e2 levels  overexpry stress . Transget stress . Moreovetomatoes . Taken tThe frequency and intensity of multifactorial stresses have perturbed plant health through ROS generation. Changing climate is a major challenge for food security, particularly when considering the ever-growing world population. Furthermore, ROS are a double-edged sword. Basal concentrations activate signaling pathways that sustain cellular homeostasis and help plants adapt to diverse stresses; however, when the concentration of ROS surpasses a particular threshold, oxidative stress is triggered. Antioxidants are essential to maintain the equilibrium between the generation of ROS and their quenching, reducing the impact of stress. Inventing techniques that prevent the ROS-induced losses during stress and elucidating the underlying mechanism will contribute to the development of crops adapted to environmental conditions. Recently, molecular priming has shown huge potential to enhance plant tolerance against environmental stresses; however, some gaps in knowledge still exist. Furthermore, identification of putative candidate genes, proteins, and metabolites regulating diverse signaling pathways using the “-omics” approach will help find new approaches . Remarka