Promoter variations of ClERF1 gene determines flesh firmness in watermelon.

BACKGROUND: Flesh firmness is a critical factor that influences fruit storability, shelf-life and consumer's preference as well. However, less is known about the key genetic factors that are associated with flesh firmness in fresh fruits like watermelon. RESULTS: In this study, through bulk segregant analysis (BSA-seq), we identified a quantitative trait locus (QTL) that influenced variations in flesh firmness among recombinant inbred lines (RIL) developed from cross between the Citrullus mucosospermus accession ZJU152 with hard-flesh and Citrullus lanatus accession ZJU163 with soft-flesh. Fine mapping and sequence variations analyses revealed that ethylene-responsive factor 1 (ClERF1) was the most likely candidate gene for watermelon flesh firmness. Furthermore, several variations existed in the promoter region between ClERF1 of two parents, and significantly higher expressions of ClERF1 were found in hard-flesh ZJU152 compared with soft-flesh ZJU163 at key developmental stages. DUAL-LUC and GUS assays suggested much stronger promoter activity in ZJU152 over ZJU163. In addition, the kompetitive allele-specific PCR (KASP) genotyping datasets of RIL populations and germplasm accessions further supported ClERF1 as a possible candidate gene for fruit flesh firmness variability and the hard-flesh genotype might only exist in wild species C. mucosospermus. Through yeast one-hybrid (Y1H) and dual luciferase assay, we found that ClERF1 could directly bind to the promoters of auxin-responsive protein (ClAux/IAA) and exostosin family protein (ClEXT) and positively regulated their expressions influencing fruit ripening and cell wall biosynthesis. CONCLUSIONS: Our results indicate that ClERF1 encoding an ethylene-responsive factor 1 is associated with flesh firmness in watermelon and provide mechanistic insight into the regulation of flesh firmness, and the ClERF1 gene is potentially applicable to the molecular improvement of fruit-flesh firmness by design breeding.

Antiproliferative and immunomodulative potential of Citrullus colocynthis and its bioactive compounds in human lymphocytes and lung cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Citrullus colocynthis (L.) Schrad is a member of the Cucurbitaceae plant family which has been used in traditional medicine for the treatment of lung diseases such as asthma and bronchitis. AIM OF THE STUDY: The study was conducted to investigate antiproliferative and immunomodulating effects of C. colocynthis and isolated cucurbitacins on human T lymphocytes and lung epithelial cells in order to evaluate their potential in the treatment of airway diseases. MATERIALS AND METHODS: Different concentrations of an ethanolic extract of C. colocynthis fruits and cucurbitacins B (CuB), E (CuE) and E-glucopyranoside (CuE-Glu) were analysed for their cytotoxicity and immunomodulatory potential on Peripheral Blood Mononuclear Cells (PBMCs) of healthy donors and on the epithelial lung cancer cell line A549. Viability and proliferation were tested using WST1 and CFSE assays. Flow cytometric analysis of AnnexinV/PI staining was used to investigate cell death through apoptosis/necrosis. Effects on regulatory mechanisms of T lymphocytes, such as CD69 and CD25 marker activation, cytokine production of the cytokines interleukin 2 (IL2), tumor necrosis factor α (TNFα) and interferon γ (IFNy) were also analysed via flow cytometry. Influences on the activator protein 1 (AP1), nuclear factor of activated T-cells (NFAT) or nuclear factor 'kappa-light-chain-enhancer' of activated B-cells (NFκB) pathways were analysed in the Jurkat reporter cell line. Cytokine secretion in A549 cells stimulated with virus-like particles was analysed using the bead-based Legendplex™ assay. RESULTS: Non-toxic concentrations of C. colocynthis and CuE-Glu showed dose-dependent effects on viability and proliferation in both T lymphocytes and A549 cells. The extracts inhibited lymphocyte activation and suppressed T cell effector functions, which was also shown by lower production of cytokines IL2, TNFα and IFNy. A dose dependent inhibition of the pathways NFκB, NFAT and AP1 in Jurkat cells could be observed. In A549 cells, especially CuE and CuE-Glu showed inhibitory effects on cytokine production following a simulated viral infection. Unglycosylated cucurbitacins were more effective in suppressing the immune function in lymphocytes than glycosylated cucurbitacins, however this activity is limited to cytotoxic concentrations. CONCLUSION: In our study we could confirm the immunmodulating effect of C. colocynthis and cucurbitacins B, E and E-glucopyranoside in vitro by suppression of different pathways of inflammation and T cell proliferation. Activity in a lung cell model using a virus-like stimulation shows promise for further research regarding cucurbitacins in airway diseases.

Streptomyces-triggered coordination between rhizosphere microbiomes and plant transcriptome enables watermelon Fusarium wilt resistance.

The use of microbial inoculant is a promising strategy to improve plant health, but their efficiency often faces challenges due to difficulties in successful microbial colonization in soil environments. To this end, the application of biostimulation products derived from microbes is expected to resolve these barriers via direct interactions with plants or soil pathogens. However, their effectiveness and mechanisms for promoting plant growth and disease resistance remain elusive. In this study, we showed that root irrigation with the extracts of Streptomyces ahygroscopicus strain 769 (S769) solid fermentation products significantly reduced watermelon Fusarium wilt disease incidence by 30% and increased the plant biomass by 150% at a fruiting stage in a continuous cropping field. S769 treatment led to substantial changes in both bacterial and fungal community compositions, and induced a highly interconnected microbial association network in the rhizosphere. The root transcriptome analysis further suggested that S769 treatment significantly improved the expression of the MAPK signalling pathway, plant hormone signal transduction and plant-pathogen interactions, particular those genes related to PR-1 and ethylene, as well as genes associated with auxin production and reception. Together, our study provides mechanistic and empirical evidences for the biostimulation products benefiting plant health through coordinating plant and rhizosphere microbiome interaction.

Mapping and validation of Fusarium wilt race 2 resistance QTL from Citrullus amarus line USVL246-FR2.

KEY MESSAGE: Fon race 2 resistant QTLs were identified on chromosomes 8 and 9. Families homozygous for resistance alleles at a haplotype of three KASP markers had 42% lower disease severity than those with susceptible alleles in an independent, interspecific validation population confirming their utility for introgression of Fusarium wilt resistance. Fusarium oxysporum f. sp. niveum (Fon) race 2 causes Fusarium wilt in watermelon and threatens watermelon production worldwide. Chemical management options are not effective, and no resistant edible watermelon cultivars have been released. Implementation of marker-assisted selection to develop resistant cultivars requires identifying sources of resistance and the underlying quantitative trait loci (QTL), developing molecular markers associated with the QTL, and validating marker-phenotype associations with an independent population. An intraspecific Citrullus amarus recombinant inbred line population from a cross of resistant USVL246-FR2 and susceptible USVL114 was used for mapping Fon race 2 resistance QTL. KASP markers were developed (N = 51) for the major QTL on chromosome 9 and minor QTL on chromosomes 1, 6, and 8. An interspecific F2:3 population was developed from resistance donor USVL246-FR2 (C. amarus) and a susceptible cultivar 'Sugar Baby' (Citrullus lanatus) to validate the utility of the markers for introgression of resistance from the wild crop relative into cultivated watermelon. Only 16 KASP markers segregated in the interspecific C. amarus/lanatus validation population. Four markers showed significant differences in the separation of genotypes based on family-mean disease severity, but together explained only 16% of the phenotypic variance. Genotypes that inherited homozygous resistant parental alleles at three KASP markers had 42% lower family-mean disease severity than homozygous susceptible genotypes. Thus, haplotype analysis was more effective at predicting the mean disease severity of families than single markers. The haplotype identified in this study will be valuable for developing Fon race 2 resistant watermelon cultivars.

The SUMOylation pathway regulates the pathogenicity of Fusarium oxysporum f. sp. niveum in watermelon through stabilizing the pH regulator FonPalC via SUMOylation.

SUMOylation is a key post-translational modification, where small ubiquitin-related modifier (SUMO) proteins regulate crucial biological processes, including pathogenesis, in phytopathogenic fungi. Here, we investigated the function and mechanism of the SUMOylation pathway in the pathogenicity of Fusarium oxysporum f. sp. niveum (Fon), the fungal pathogen that causes watermelon Fusarium wilt. Disruption of key SUMOylation pathway genes, FonSMT3, FonAOS1, FonUBC9, and FonMMS21, significantly reduced pathogenicity, impaired penetration ability, and attenuated invasive growth capacity of Fon. Transcription and proteomic analyses identified a diverse set of SUMOylation-regulated differentially expressed genes and putative FonSMT3-targeted proteins, which are predicted to be involved in infection, DNA damage repair, programmed cell death, reproduction, growth, and development. Among 155 putative FonSMT3-targeted proteins, FonPalC, a Pal/Rim-pH signaling regulator, was confirmed to be SUMOylated. The FonPalC protein accumulation was significantly decreased in SUMOylation-deficient mutant ∆Fonsmt3. Deletion of FonPalC resulted in impaired mycelial growth, decreased pathogenicity, enhanced osmosensitivity, and increased intracellular vacuolation in Fon. Importantly, mutations in conserved SUMOylation sites of FonPalC failed to restore the defects in ∆Fonpalc mutant, indicating the critical function of the SUMOylation in FonPalC stability and Fon pathogenicity. Identifying key SUMOylation-regulated pathogenicity-related proteins provides novel insights into the molecular mechanisms underlying Fon pathogenesis regulated by SUMOylation.

Fine mapping of ClLOX, a QTL for powdery mildew resistance in watermelon (Citrullus lanatus L.).

KEY MESSAGE: ClLOX, is located on chromosome 2 and encodes a lipoxygenase gene, which induced watermelon powdery mildew resistance by inhibiting pathogen spread. Powdery mildew is one of the most severe fungal diseases reducing yield and quality of watermelon (Citrullus lanatus L.) and other cucurbit crops. Genes responsible for powdery mildew resistance in watermelon are highly valuable. In this study, we first identified the QTL pm-lox for powdery mildew resistance in watermelon, located within a 0.93 Mb interval of chromosome 2, via XP-GWAS method using two F2 populations. The F2:3 families from one of the F2 populations were then used for fine-mapping the pm-lox locus into a 9,883 bp physical region between 29,581,906 and 29,591,789, containing only two annotated genes. Of these, only ClG42_02g0161300 showed a significant differential expression between the resistant and susceptible lines after powdery mildew inoculation based on RNA sequencing (RNA-seq) and qRT-PCR analysis, and is designated ClLOX. Derived Cleaved Amplified Polymorphic Sequence (dCAPs) markers were developed and validated. In addition, our tests showed that the resistance was anti-spread rather than anti-infection of the pathogen. This study identified a new resistance gene (ClLOX), provided insights into the mechanism of powdery mildew resistance, and developed a molecular marker for watermelon breeding.

A CRISPR/LbCas12a-based method for detection of bacterial fruit blotch pathogens in watermelon.

Acidovorax citrulli is the main pathogen causing bacterial fruit blotch, which seriously threatens the global watermelon industry. At present, rapid, sensitive, and low-cost detection methods are urgently needed. The established CRISPR/LbCas12a visual detection method can specifically detect A. citrulli and does not cross-react with other pathogenic bacteria such as Erwinia tracheiphila, Pseudomonas syringae, and Xanthomonas campestris. The sensitivity of this method for genomic DNA detection is as low as 0.7 copies/µL, which is higher than conventional PCR and real-time PCR. In addition, this method only takes 2.5 h from DNA extraction to quantitative detection and does not require complex operation and sample treatment. Additionally, the technique was applied to test real watermelon seed samples for A. citrulli, and the results were contrasted with those of real-time fluorescence quantitative PCR and conventional PCR. The high sensitivity and specificity have broad application prospects in the rapid detection of bacterial fruit blotch bacterial pathogens of watermelon.IMPORTANCEBacterial fruit blotch, Acidovorax citrulli, is an important seed-borne bacterial disease of watermelon, melon, and other cucurbits. The lack of rapid, sensitive, and reliable pathogen detection methods has hampered research on fruit spot disease prevention and control. Here, we demonstrate the CRISPR/Cas12a system to analyze aspects of the specificity and sensitivity of A. citrulli and to test actual watermelon seed samples. The results showed that the CRISPR/Cas12a-based free-amplification method for detecting bacterial fruit blotch pathogens of watermelons was specific for A. citrulli target genes and 100-fold more sensitive than conventional PCR with quantitative real-time PCR. This method provides a new technical tool for the detection of A. citrulli.

Organic fertilizer type and dose affect growth, morphological and physiological parameters, and mineral nutrition of watermelon seedlings.

Background: Organic agriculture has grown rapidly in recent years due to its environmental friendliness, sustainability, and improved farm profitability. Transplants are commonly used for fruits and vegetables to achieve consistent quality, uniformity, and easy field spacing control. The efficacy and optimal amounts of fertilizers for organic transplant production need to be investigated. Methods: The effects of three organic fertilizers (Sustane 4-6-4, Nature Safe 7-7-7, and Dramatic 2-4-1) and one conventional fertilizer Peters Professional 20-20-20 (Conventional) with four doses (nitrogen (N) content was matched among fertilizers in each level, as 0.14 g/L, 0.28 g/L, 0.56 g/L, and 0.84 g/L N, respectively) on watermelon seedlings were compared in this study. Results: The results showed that all organic fertilizer treatments were not significantly different from the Conventional group in terms of watermelon germination. The only exception was the highest dose of Sustane 4-6-4 (0.84 g/L N) which decreased the germination rate and relative emergence index. Generally, growth index, shoot fresh and dry weights, true leaf number, and stem diameter increased as the amount of N increased within each fertilizer type. The best shoot growth was observed in the highest doses of Conventional and Dramatic 2-4-1 treatments (0.84 g/L N). However, Dramatic 2-4-1 treatments resulted in the lowest root growth when compared to other fertilizers at the same N dose. The second highest fertilization dose (0.56 g/L N) of Sustane 4-6-4 had the best root growth according to root fresh weight, root volume, root area, total root length, as well as the numbers of root tip and crossing when compared to other treatments. For seedlings, a well-developed root system can ensure a good seedling establishment and high survival rate under stressful field conditions after transplanting. Thus, Sustane 4-6-4 at 14 g/L (0.56 g/L N) is recommended to produce high-quality organic watermelon seedlings among the treatments applied in this study.

Impact of sulfite use and acidification on chemical quality components in thermally processed watermelon juices.

The present study compared pasteurized and reconstituted (from vacuum-concentrated) watermelon juices with sulfite use (∼40 mg/L) and acidification (pH = 4.2) to fresh watermelon juices. The products were evaluated for phenolics, free amino acids, carotenoids, sugars, organic acids, and alcohols by high-performance liquid chromatography-HPLC and the volatile profile by headspace-gas chromatography/mass spectrometry(HS-GC/MS). Pasteurization had no significant impact on most of the chemical components. Furthermore, it potentiated typical watermelon aromas (E,E)-2,6-nonadienal, (Z)-3-nonen-1-ol, 4-hexen-1-ol, (E,Z)-3,6-nonadien-1-ol, 6-amino-2-methyl-2-heptanol, (E)-6-nonenal, (E)-2-nonenal, pentanal, nonanal and 1-nonanol), without off-flavor compounds formation. On the other hand, the reconstituted juice showed reduced amino acids (serine, glutamine, and tryptophan), phenolics (epicatechin gallate, myricetin, and cis-resveratrol), carotenoids (lycopene, ß-carotene, and violaxanthin) and most volatile compounds. Our results showed that sulfite and acidification could maintain watermelon juice's nutritional and quality parameters after pasteurization. The vacuum concentration and reconstitution processes negatively impacted the evaluated compounds. Our findings contribute to improving thermal processes in watermelon juices for better preservation of nutrients, flavor, and bioactive compounds.

Genome-Wide Identification and Expression Analysis of Chitinase Genes in Watermelon under Abiotic Stimuli and Fusarium oxysporum Infection.

Chitinases, which catalyze the hydrolysis of chitin, the primary components of fungal cell walls, play key roles in defense responses, symbiotic associations, plant growth, and stress tolerance. In this study, 23 chitinase genes were identified in watermelon (Citrullus lanatus [Thunb.]) and classified into five classes through homology search and phylogenetic analysis. The genes with similar exon-intron structures and conserved domains were clustered into the same class. The putative cis-elements involved in the responses to phytohormone, stress, and plant development were identified in their promoter regions. A tissue-specific expression analysis showed that the ClChi genes were primarily expressed in the roots (52.17%), leaves (26.09%), and flowers (34.78%). Moreover, qRT-PCR results indicate that ClChis play multifaceted roles in the interaction between plant/environment. More ClChi members were induced by Race 2 of Fusarium oxysporum f. sp. niveum, and eight genes were expressed at higher levels on the seventh day after inoculation with Races 1 and 2, suggesting that these genes play a key role in the resistance of watermelon to Fusarium wilt. Collectively, these results improve knowledge of the chitinase gene family in watermelon species and help to elucidate the roles played by chitinases in the responses of watermelon to various stresses.

Specific antibody production using recombinant proteins to elucidate seed transmission and nuclear localization of Coguvirus citrulli and Coguvirus henanense in radicles of watermelon crop.

Watermelon crinkle leaf-associated virus 1 (WCLaV-1) and WCLaV-2, both belonging to the genus Coguvirus (family Phenuiviridae), have been identified in watermelon plants in Brazil. To study tissue tropism and the potential for seed transmission of these viruses, we initially planned to produce specific antibodies. However, difficulties in isolating and propagating the virus in host plants hindered the purified virus preparations. To overcome this problem, the nucleocapsid (N) proteins of WCLaV-1 and -2 were produced using the pepper ringspot virus vector. The N protein genes and the vector backbone were prepared by (RT-)PCR and ligated by Gibson assembly. The constructs were agro-infiltrated in Nicotiana benthamiana plants. The expressed N proteins were purified and used for polyclonal antibody production. The specificity of both antibodies was confirmed by antigen-coating ELISA, tissue-blot immunobinding assay and Western blot. By antigen-coating ELISA demonstrated that WCLaV-1 showed 93.1% of seed-transmission, while WCLaV-2 showed only 17.8%. The N protein of WCLaV-1 was detected in the cytoplasm of the seed tissues. It was also found in the nuclei of the radicle, as confirmed by confocal microscopy. We concluded that the antibodies exhibited both a high titer and sufficient specificity for use in ELISA-based diagnostics and for subcellular localization study.

A natural substitution of a conserved amino acid in eIF4E confers resistance against multiple potyviruses.

Eukaryotic translation initiation factor 4E (eIF4E), which plays a pivotal role in initiating translation in eukaryotic organisms, is often hijacked by the viral genome-linked protein to facilitate the infection of potyviruses. In this study, we found that the naturally occurring amino acid substitution D71G in eIF4E is widely present in potyvirus-resistant watermelon accessions and disrupts the interaction between watermelon eIF4E and viral genome-linked protein of papaya ringspot virus-watermelon strain, zucchini yellow mosaic virus or watermelon mosaic virus. Multiple sequence alignment and protein modelling showed that the amino acid residue D71 located in the cap-binding pocket of eIF4E is strictly conserved in many plant species. The mutation D71G in watermelon eIF4E conferred resistance against papaya ringspot virus-watermelon strain and zucchini yellow mosaic virus, and the equivalent mutation D55G in tobacco eIF4E conferred resistance to potato virus Y. Therefore, our finding provides a potential precise target for breeding plants resistant to multiple potyviruses.

Consumer perception of whole watermelons.

There are many varieties of watermelons, providing distinct external and internal sensory attributes. This study used an online survey (n = 700) and focus groups (n = 25) to investigate consumer perception of whole watermelons. Rind color, sound of the melon, size, and price were the most important attributes for consumers when selecting a whole watermelon. Freshness was the most important whole watermelon characteristic, and watermelon freshness/quality was driven by sweetness, crispness, and juiciness. Consumers preferred seedless watermelons that had a light rind with dark green stripes, red flesh, an oval/oblong shape, firm and crisp flesh, a weight of approximately 2.2-5.5 kg, and labeling that described them as fresh, juicy, and sweet. Two consumer clusters were identified from quantitative survey data and were also representative of focus group participants: value consumers and watermelon enthusiasts. Watermelon enthusiasts were differentiated by a higher value for claims including local, product of USA, sustainably farmed, and organic. Watermelon purchase is quality driven: consumers will pay more for guaranteed sweetness and crispness. PRACTICAL APPLICATION: The ideal watermelon for all consumers is one that is dark green with stripes, is medium sized and oblong in shape, has a minimal rind-to-flesh ratio, and boasts dark, vibrant red flesh that is sweet, crisp, and juicy. All consumers want a better guarantee on watermelon quality because it is hard to predict sensory quality when selecting a melon. This study demonstrated the intrinsic and external drivers of liking for fresh watermelons and summarized a consumer watermelon purchase and consumption journey map that can guide further research and development of watermelons and provide insights on how to increase watermelon sales.

Effective application of citric acid treated Trapa natans and Citrullus lanatus lignocellulosic macromolecules for adsorptive remediation of acid Violet-7 dye.

The peels of Trapa natans (TRA) and Citrullus lanatus (CIT), were modified with a variety of chemicals to boost their surface for the optimization of adsorption performance by providing a greater number of additional active binding sites. Citric acid-processed peels (TRAC and CITC) had shown more favorable adsorption performance to eradicate acid violet 7 dye (AVS). Extra and additional active sites generated after chemical processing, including hydroxyl (OH), carboxyl (COOH), amines NH2, carbonyl, and ester (-O-CO-) groups, as evidenced from FTIR and SEM characterizations, may boost the potential of physicochemical integration of adsorbent surface activity in order to promote and encourage the retention of hazardous and risky AVS molecules from the water. The Langmuir isotherm assessed the qmax for the adsorption of AVS on TRAC, CITC, TRA, and CIT to be 212.8, 294, 24.3, and 60.6 mg/g, respectively, whereas the correlation coefficients assessed for both TRAC and CITC were 0.98 and for TRA and CIT were 0.97, closer to unity reflecting monolayer physio-sorption. According to Temkin, the adsorption of AVS on TRAC, TRA, CITC, and CIT gives "BT" values of 1.275, 0.947, 1.085, and 1.211 mg/g, also suggesting physio-sorption. Therefore, chemically modified peels can be employed for detoxification of AVS.

Bacillus velezensis WB induces systemic resistance in watermelon against Fusarium wilt.

BACKGROUND: Our previous findings indicated that Bacillus velezensis WB could control Fusarium wilt by changing the structure of the microbial community in the watermelon rhizosphere. However, there are few studies on its mechanism in the pathogen resistance of watermelon. Therefore, in this study, we determined the mechanism of B. velezensis WB-induced systemic resistance in watermelon against Fusarium wilt through glasshouse pot experiments. RESULTS: The results showed that B. velezensis WB significantly reduced the incidence and disease index of Fusarium wilt in watermelon. B. velezensis WB can enhance the basal immunity of watermelon plants by: increasing the activity of phenylalanine ammonia-lyase (PAL), peroxidase (POD), superoxide dismutase (SOD) and ß-1,3-glucanase; accumulating lignin, salicylic acid (SA) and jasmonic acid (JA); reducing malondialdehyde (MDA) concentrations; and inducing callus deposition in watermelon plant cells. RNA-seq analysis showed that 846 watermelon genes were upregulated and 612 watermelon genes were downregulated in the WF treatment. This process led to the activation of watermelon genes associated with auxin, gibberellin, SA, ethylene and JA, and the expression of genes in the phenylalanine biosynthetic pathway was upregulated. In addition, transcription factors involved in plant resistance to pathogens, such as MYB, NAC and WRKY, were induced. Gene correlation analysis showed that Cla97C10G195840 and Cla97C02G049930 in the phenylalanine biosynthetic pathway, and Cla97C02G041360 and Cla97C10G197290 in the plant hormone signal transduction pathway showed strong correlations with other genes. CONCLUSION: Our results indicated that B. velezensis WB is capable of inducing systemic resistance in watermelon against Fusarium wilt. © 2023 Society of Chemical Industry.

Natural variation in a short region of the Acidovorax citrulli type III-secreted effector AopW1 is associated with differences in cytotoxicity and host adaptation.

Bacterial fruit blotch, caused by Acidovorax citrulli, is a serious disease of melon and watermelon. The strains of the pathogen belong to two major genetic groups: group I strains are strongly associated with melon, while group II strains are more aggressive on watermelon. A. citrulli secretes many protein effectors to the host cell via the type III secretion system. Here we characterized AopW1, an effector that shares similarity to the actin cytoskeleton-disrupting effector HopW1 of Pseudomonas syringae and with effectors from other plant-pathogenic bacterial species. AopW1 has a highly variable region (HVR) within amino acid positions 147 to 192, showing 14 amino acid differences between group I and II variants. We show that group I AopW1 is more toxic to yeast and Nicotiana benthamiana cells than group II AopW1, having stronger actin filament disruption activity, and increased ability to induce cell death and reduce callose deposition. We further demonstrated the importance of some amino acid positions within the HVR for AopW1 cytotoxicity. Cellular analyses revealed that AopW1 also localizes to the endoplasmic reticulum, chloroplasts, and plant endosomes. We also show that overexpression of the endosome-associated protein EHD1 attenuates AopW1-induced cell death and increases defense responses. Finally, we show that sequence variation in AopW1 plays a significant role in the adaptation of group I and II strains to their preferred hosts, melon and watermelon, respectively. This study provides new insights into the HopW1 family of bacterial effectors and provides first evidence on the involvement of EHD1 in response to biotic stress.

Removal of dyes from water using Citrullus lanatus seed powder in continuous and discontinuous systems.

The objective of this study is to develop a low-cost biosorbent using residual seeds of the Citrullus lanatus fruit for the removal of cationic dyes. Physicochemical parameters such as pH, adsorbent mass, contact time, and temperature were evaluated for their effects on dye removal. The biosorbent is composed of lignin and cellulose, exhibiting a highly heterogeneous surface with randomly distributed cavities and bulges. The adsorption of both dyes was most effective at natural pH with a dosage of 0.8 g L-1. Equilibrium was reached within 120 min, regardless of concentration, indicating rapid kinetics. The Elovich model and pseudo-second-order kinetics were observed for crystal violet and basic fuchsin dye, respectively. The Langmuir model fitted well with the equilibrium data of both dyes. However, the increased temperature had a negative impact on dye adsorption. The biosorbent also demonstrated satisfactory performance (R = 43%) against a synthetic mixture of dyes and inorganic salts, with a small mass transfer zone. The adsorption capacities for crystal violet and basic fuchsin dye were 48.13 mg g-1 and 44.26 mg g-1, respectively. Thermodynamic studies confirmed an exothermic nature of adsorption. Overall, this low-cost biosorbent showed potential for the removal of dyes from aqueous solutions.

In this work, a novel biosorbent was developed using residual Citrullus lanatus fruit seeds that can efficiently remove cationic dyes from aqueous solutions. The biosorbent's composition includes lignin and cellulose, and its surface structure is highly heterogeneous, consisting of randomly distributed cavities and bulges. The biosorbent demonstrated a rapid and efficient adsorption capacity for both crystal violet and basic fuchsin, regardless of dye concentration. Moreover, the biosorbent was successfully employed in the treatment of a synthetic mixture containing several dyes and inorganic salts. Finally, the application of the biosorbent in continuous adsorption showed a low zone of mass transfer and high breakthrough time, indicating it to be an excellent material for fixed-bed operation. Overall, this study provides a low-cost and efficient alternative for the removal of dyes from aqueous solutions, with promising practical applications.

Transcriptome and Metabolome Analyses Reveal That Jasmonic Acids May Facilitate the Infection of Cucumber Green Mottle Mosaic Virus in Bottle Gourd.

Cucumber green mottle mosaic virus (CGMMV) is a typical seed-borne tobamovirus that mainly infects cucurbit crops. Due to the rapid growth of international trade, CGMMV has spread worldwide and become a significant threat to cucurbit industry. Despite various studies focusing on the interaction between CGMMV and host plants, the molecular mechanism of CGMMV infection is still unclear. In this study, we utilized transcriptome and metabolome analyses to investigate the antiviral response of bottle gourd (Lagenaria siceraria) under CGMMV stress. The transcriptome analysis revealed that in comparison to mock-inoculated bottle gourd, 1929 differently expressed genes (DEGs) were identified in CGMMV-inoculated bottle gourd. Among them, 1397 genes were upregulated while 532 genes were downregulated. KEGG pathway enrichment indicated that the DEGs were mainly involved in pathways including the metabolic pathway, the biosynthesis of secondary metabolites, plant hormone signal transduction, plant-pathogen interaction, and starch and sucrose metabolism. The metabolome result showed that there were 76 differentially accumulated metabolites (DAMs), of which 69 metabolites were up-accumulated, and 7 metabolites were down-accumulated. These DAMs were clustered into several pathways, including biosynthesis of secondary metabolites, tyrosine metabolism, flavonoid biosynthesis, carbon metabolism, and plant hormone signal transduction. Combining the transcriptome and metabolome results, the genes and metabolites involved in the jasmonic acid and its derivatives (JAs) synthesis pathway were significantly induced upon CGMMV infection. The silencing of the allene oxide synthase (AOS) gene, which is the key gene involved in JAs synthesis, reduced CGMMV accumulation. These findings suggest that JAs may facilitate CGMMV infection in bottle gourd.

Irrigation with saline water in the cultivation of mini watermelon under phosphate fertilization.

The objective of this study was to evaluate the water status, photosynthetic pigments, and photochemical efficiency of mini watermelon plants under salt stress and phosphate fertilization. The experiment was conducted in pots under greenhouse conditions in Pombal, PB, Brazil. The experimental design used was randomized blocks in a 5 × 4 factorial scheme, with five levels of electrical conductivity of irrigation water - ECw (0.3, 1.3, 2.3, 3.3, and 4.3 dS m-1) and four doses of phosphorus (60, 80, 100, and 120% of the recommendation), with three replicates. The relative water content in the tissues decreased with the increase in ECw levels in all phosphorus doses, with decreases of 7.05, 7.81 and 8.83% per unit increase in ECw, in plants fertilized with 80, 100 and 120% P2O5. On the other hand, ECw levels increased electrolyte leakage, regardless of phosphorus doses of the recommendation. The synthesis of photosynthetic pigments and the quantum efficiency of photosystem II were inhibited by increasing water salinity from 0.3 dS m-1 in plants grown under phosphorus doses above 60% of the recommendation. Water salinity from 0.3 dS m-1 reduced chlorophyll b contents, initial, maximum, and variable fluorescence of mini watermelon plants, with a decrease of 11.86, 4.51, 4.53, and 4.54% per unit increment of ECw, respectively.

From Fruit Waste to Medical Insight: The Comprehensive Role of Watermelon Rind Extract on Renal Adenocarcinoma Cellular and Transcriptomic Dynamics.

Cancer researchers are fascinated by the chemistry of diverse natural products that show exciting potential as anticancer agents. In this study, we aimed to investigate the anticancer properties of watermelon rind extract (WRE) by examining its effects on cell proliferation, apoptosis, senescence, and global gene expression in human renal cell adenocarcinoma cells (HRAC-769-P) in vitro. Our metabolome data analysis of WRE exhibited untargeted phyto-constituents and targeted citrulline (22.29 µg/mg). HRAC-769-P cells were cultured in RPMI-1640 media and treated with 22.4, 44.8, 67.2, 88.6, 112, 134.4, and 156.8 mg·mL-1 for 24, 48, and 72 h. At 24 h after treatment, (88.6 mg·mL-1 of WRE) cell proliferation significantly reduced, more than 34% compared with the control. Cell viability decreased 48 and 72 h after treatment to 45% and 37%, respectively. We also examined poly caspase, SA-beta-galactosidase (SA-beta-gal), and wound healing activities using WRE. All treatments induced an early poly caspase response and a significant reduction in cell migration. Further, we analyzed the transcript profile of the cells grown at 44.8 mg·mL-1 of WRE after 6 h using RNA sequencing (RNAseq) analysis. We identified 186 differentially expressed genes (DEGs), including 149 upregulated genes and 37 downregulated genes, in cells treated with WRE compared with the control. The differentially expressed genes were associated with NF-Kappa B signaling and TNF pathways. Crucial apoptosis-related genes such as BMF, NPTX1, NFKBIA, NFKBIE, and NFKBID might induce intrinsic and extrinsic apoptosis. Another possible mechanism is a high quantity of citrulline may lead to induction of apoptosis by the production of increased nitric oxide. Hence, our study suggests the potential anticancer properties of WRE and provides insights into its effects on cellular processes and gene expression in HRAC-769-P cells.