Fusarium Wilt Of Watermelon Research Articles

The secreted protein FonCHRD is essential for vegetative growth, asexual reproduction, and pathogenicity in watermelon Fusarium wilt fungus

Fungal pathogens often secrete numerous effectors to interfere with and/or suppress plant immunity to promote their infection. Watermelon Fusarium wilt, caused by the soil-borne fungus Fusarium oxysporum f. sp. niveum (Fon), is one of the devastating diseases that severely affect the watermelon industry. Here, we report the function of a candidate effector protein, FonCHRD, in Fon. FonCHRD harbors a chordin (CHRD) domain of unknown function and has a signal peptide with secretion activity. FonCHRD shows a relatively high expression level in Fon marcoconidia and is inducible by watermelon root tissues. Phenotypic analysis of the targeted deletion mutant revealed that FonCHRD plays roles in vegetative growth, asexual reproduction, and conidial morphology of Fon, while it is not involved in spore germination as well as cell wall, oxidative and salt stress responses. Deletion of FonCHRD impaired the ability to colonize and spread within host plants, significantly reducing its virulence on watermelon. FonCHRD is distributed across multiple compartments of plant cells but can target to the apoplast space in plants. FonCHRD inhibits the INF1- and Bcl2-associated X protein-triggered cell death and defense gene expression in transiently expressed Nicotiana benthamiana leaves. These findings suggest that FonCHRD is essential for Fon pathogenicity by modulating invasive growth and spreading abilities as well as by suppressing plant immune responses.

Production-optimized fermentation of antifungal compounds by bacillus velezensis LZN01 and transcriptome analysis.

Fusarium wilt is one of the major constraints on global watermelon production, and Fusarium oxysporum f. sp. niveum (Fon) is the causative agent of Fusarium wilt in watermelon and results in severe yield and quality losses worldwide. The enhancement of antifungal activity from antagonistic bacteria against Fon is highly practical for managing Fusarium wilt in watermelon. The aim of this study was to maximize the antifungal activity of Bacillus velezensis LZN01 by optimizing fermentation conditions and analysing its regulatory mechanism via transcriptome sequencing. The culture and fermentation conditions for strain LZN01 were optimized by single-factor and response surface experiments. The optimum culture conditions for this strain were as follows: the addition of D-fructose at 35 g/L and NH4Cl at 5 g/L in LB medium, pH 7, and incubation at 30°C for 72 h. The fungal inhibition rate for strain LZN01 reached 71.1%. The improvement of inhibition rate for strain LZN01 in optimization fermentation was supported by transcriptomic analysis; a total of 491 genes were upregulated, while 736 genes were downregulated. Transcriptome analysis revealed that some differentially expressed genes involved in carbon and nitrogen metabolism, oxidation-reduction, fatty acid and secondary metabolism; This optimization process could potentially lead to significant alterations in the production levels and types of antimicrobial compounds by the strain. Metabolomics and UPLC/Q-Exactive Orbitrap MS analysis revealed that the production yields of antimicrobial compounds, such as surfactin, fengycin, shikimic acid, and myriocin, increased or were detected in the cell-free supernatant (CFS) after the fermentation optimization process. Our results indicate that fermentation optimization enhances the antifungal activity of the LZN01 strain by influencing the expression of genes responsible for the synthesis of antimicrobial compounds.

Antagonistic effect of Bacillus and Pseudomonas combinations against Fusarium oxysporum and their effect on disease resistance and growth promotion in watermelon.

We aimed to develop an effective bacterial combination that can combat Fusarium oxysporum infection in watermelon using in vitro and pot experiments. In total, 53 strains of Bacillus and 4 strains of Pseudomonas were screened. Pseudomonas strains P3 and P4 and Bacillus strains XY-2-3, XY-13, and GJ-1-15 exhibited good antagonistic effects against F. oxysporum. P3 and P4 were identified as Pseudomonas chlororaphis and Pseudomonas fluorescens, respectively. XY-2-3 and GJ-1-15 were identified as B. velezensis, and XY-13 was identified as Bacillus amyloliquefaciens. The three Bacillus strains were antifungal, promoted the growth of watermelon seedlings and had genes to synthesize antagonistic metabolites such as bacilysin, surfactin, yndj, fengycin, iturin, and bacillomycin D. Combinations of Bacillus and Pseudomonas strains, namely, XY-2-3+P4, GJ-1-15+P4, XY-13+P3, and XY-13+P4, exhibited a good compatibility. These four combinations exhibited antagonistic effects against 11 pathogenic fungi, including various strains of F. oxysporum, Fusarium solani, and Rhizoctonia. Inoculation of these bacterial combinations significantly reduced the incidence of Fusarium wilt in watermelon, promoted plant growth, and improved soil nutrient availability. XY-13+P4 was the most effective combination against Fusarium wilt in watermelon with the inhibition rate of 78.17%. The number of leaves; aboveground fresh and dry weights; chlorophyll, soil total nitrogen, and soil available phosphorus content increased by 26.8%, 72.12%, 60.47%, 16.97%, 20.16%, and 16.50%, respectively, after XY-13+P4 inoculation compared with the uninoculated control. Moreover, total root length, root surface area, and root volume of watermelon seedlings were the highest after XY-13+P3 inoculation, exhibiting increases by 265.83%, 316.79%, and 390.99%, respectively, compared with the uninoculated control. XY-13+P4 was the best bacterial combination for controlling Fusarium wilt in watermelon, promoting the growth of watermelon seedlings, and improving soil nutrient availability.

The Ser/Thr protein kinase FonKin4-poly(ADP-ribose) polymerase FonPARP1 phosphorylation cascade is required for the pathogenicity of watermelon fusarium wilt fungus Fusarium oxysporum f. sp. niveum.

Poly(ADP-ribosyl)ation (PARylation), catalyzed by poly(ADP-ribose) polymerases (PARPs) and hydrolyzed by poly(ADP-ribose) glycohydrolase (PARG), is a kind of post-translational protein modification that is involved in various cellular processes in fungi, plants, and mammals. However, the function of PARPs in plant pathogenic fungi remains unknown. The present study investigated the roles and mechanisms of FonPARP1 in watermelon Fusarium wilt fungus Fusarium oxysporum f. sp. niveum (Fon). Fon has a single PARP FonPARP1 and one PARG FonPARG1. FonPARP1 is an active PARP and contributes to Fon pathogenicity through regulating its invasive growth within watermelon plants, while FonPARG1 is not required for Fon pathogenicity. A serine/threonine protein kinase, FonKin4, was identified as a FonPARP1-interacting partner by LC-MS/MS. FonKin4 is required for vegetative growth, conidiation, macroconidia morphology, abiotic stress response and pathogenicity of Fon. The S_TKc domain is sufficient for both enzyme activity and pathogenicity function of FonKin4 in Fon. FonKin4 phosphorylates FonPARP1 in vitro to enhance its poly(ADP-ribose) polymerase activity; however, FonPARP1 does not PARylate FonKin4. These results establish the FonKin4-FonPARP1 phosphorylation cascade that positively contributes to Fon pathogenicity. The present study highlights the importance of PARP-catalyzed protein PARylation in regulating the pathogenicity of Fon and other plant pathogenic fungi.

Evaluation of differential miRNA expression between Fusarium wilt-resistant and -susceptible watermelon varieties

Watermelon (Citrullus lanatus) has been cultivated for nearly one thousand years and remains a commercially important crop. However, continuous planting often leads to the aggravation of many diseases. Watermelon Fusarium wilt (FW) is one of them, which is a fungal soil-borne disease caused by Fusarium oxysporum f. sp. niveum (Fon) that damages plant roots by invading vascular bundles. Therefore, it is imperative to comprehensively characterize the mechanism mediating disease resistance or susceptibility, as well as identify effective resistance genes, in order to breed improved disease-resistant watermelon varieties. miRNAs are endogenous non-coding RNAs that are widely involved in plant growth, development, metabolism, transport, stress response, and pathogen defense. However, there are few reports of disease-associated miRNAs, or their mechanisms of action, in Cucurbitaceae crops. In this study, the roots of both Fon-susceptible and resistant watermelon varieties were infected with GFP-labeled Fon. Three key infection time points were identified. Illumina sequencing was used to obtain Fon-resistance related miRNAs, and degradome sequencing was used to obtain miRNA target genes. A total of 23 differentially expressed miRNAs were identified at the three key infection time points, the degradation group sequencing found their only target gene. The results of this study provide a theoretical basis for studies of miRNA function and regulation during the interaction between Fon and watermelon plants and clues for the screening of watermelon resistance genes. In addition, this information can be used to breed disease-resistant watermelon varieties.

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

Key messageFon 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.

Management of Fusarium wilt of watermelon (Citrullus lanatus) using Bacillus subtilus and Bacillus cereus

Fusarium wilt of watermelon [Citrullus lanatus (Thunb.) Matsumura & Nakai] is caused by Fusarium oxysporum f. sp. niveum (FON), is the most severe soil-borne disease under the prevailing conditions of Punjab. Present study was carried out during 2018–19 and 2019–20 at the pot house and research farm of Punjab Agricultural University, Ludhiana, Punjab to evaluate the antagonistic activity of 15 isolates of Bacillus spp. against FON. These isolates were screened by using dual culture assay and among them, Bacillus subtilus (B1) and Bacillus cereus (B3) showed maximum inhibition percentage i.e. 57.00 and 58.22, respectively. Similarly, these isolates (B1 and B3) showed maximum activity of chitinase (2.31 and 2.16 unit/ml) and glucanase (1.72 and 1.79 unit/ml). Further, Talc-based bio- formulations of B1 and B3 isolates were evaluated in pot house and research farm of Punjab Agricultural University, Ludhiana, Punjab. Amongst all the treatments, Bacillus subtilis (B1) and Bacillus cereus (B3) as seed + soil @15 g treatment showed maximum inhibition of FON. Therefore, based on our findings, Bacillus subtilis (B1) and Bacillus cereus (B3) used as seed + soil treatment was able to effectively manage the fusarium wilt in watermelon under pothouse and field conditions.

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.

Tacrolimus analogue produced by Bacillus amyloliquefaciens HSSN09 suppresses watermelon Fusarium wilt by antagonizing FON

Watermelon Fusarium wilt caused by Fusarium oxysporum f. sp. niveum (FON) is one of the most devastating diseases in watermelon production. Biological control approaches are efficient and environmentally-friendly, thus have aroused broad interest. However, only a few biocontrol factors/molecules have been characterized due to varied genomes and diversified secreted secondary metabolites of biocontrol microorganisms. We report the screen and identification of a Bacillus amyloliquefaciens strain HSSN09 with high antagonism to FON and watermelon growth promotion capacity. B. amyloliquefaciens HSSN09 and its secondary metabolites showed inhibition on FON mycelial growth, spore formation and spore germination. Metabolomes of the highly antagonistic strain B. amyloliquefaciens HSSN09, medium antagonistic strain B. amyloliquefaciens FGW53, weakly antagonistic strain B. subtilis JC03 and non-antagonistic strain B. cereus FGW109 were compared with LC-MS/MS. By antagonistic assay, tacrolimus was screened out among the 531 molecules that appeared in the sequencing results of HSSN09 metabolome but not in that of the non-antagonistic strain without significant inhibition on the growth of FON. Among the four tested Bacillus strains, tacrolimus analogue was only observed in the supernatant of HSSN09 by HPLC and MALDI-TOF-MS. We further confirmed the ability of tacrolimus to suppress FON and promote watermelon seedling growth.

Sensitivity of Meloidogyne incognita, Fusarium oxysporum f. sp. niveum, and Stagonosporopsis citrulli to Succinate Dehydrogenase Inhibitors Used for Control of Watermelon Diseases.

Watermelon is affected by diseases such as Fusarium wilt, gummy stem blight, and root-knot nematode (RKN). Succinate dehydrogenase inhibitors (SDHIs) with potential fungicide and nematicide activity provide the opportunity to control multiple diseases with one compound. In this study, we aimed to determine the sensitivity of Meloidogyne incognita race 4 (MI4), Fusarium oxysporum f. sp. niveum (FON), and Stagonosporopsis citrulli (SCIT) to existing SDHIs: benzovindiflupyr, fluopyram, cyclobutrifluram, and pydiflumetofen. All SDHIs had fungicidal activity against 19 SCIT isolates in mycelial growth assays, but isolates were most sensitive to pydiflumetofen (median EC50 = 0.41 μg/ml). Most of the 50 FON isolates tested were sensitive to cyclobutrifluram for mycelial growth (median EC50 = 4.04 μg/ml) and conidial germination (median EC50 = 0.2 μg/ml) assays but were not sensitive to fluopyram. MI4 was most sensitive to cyclobutrifluram for egg hatch (mean EC50 = 0.0019 μg/ml) and J2 motility (mean EC50 = 1.16 μg/ml) assays but was not sensitive to pydiflumetofen. Significant positive correlations between the sensitivity of SCIT (mycelial growth) and FON (mycelial growth and conidial germination) for cyclobutrifluram and benzovindiflupyr (SCIT r = 0.88; FON r = 0.7; P < 0.0001) and cyclobutrifluram and pydiflumetofen (SCIT r = 0.83; FON r = 0.67 and 0.77; P < 0.0001) indicate a potential for cross-resistance between these SDHIs for these fungal pathogens. Overall, results suggest that cyclobutrifluram may be used for managing RKN, whereas it should be used judiciously for Fusarium wilt of watermelon and gummy stem blight due to the existence of insensitive isolates to the fungicide.

Effects of wheat intercropping on growth and occurrence of Fusarium wilt in watermelon.

Watermelon is commonly affected by Fusarium wilt in a monoculture cropping system. Wheat intercropping alleviates the affection of Fusarium wilt of watermelon. The objective of this study was to determine the effects of wheat and watermelon intercropping on watermelon growth and Fusarium wilt. Our results showed that wheat and watermelon intercropping promoted growth, increased chlorophyll content, and photosynthesis of watermelon. Meanwhile, wheat and watermelon intercropping inhibited watermelon Fusarium wilt occurrence, decreased spore numbers, increased root vigor, increased antioxidant enzyme activities, and decreased malondialdehyde (MDA) content in watermelon roots. Additionally, wheat and watermelon intercropping enhanced the bacterial colonies and total microbes growth in soil, decreased fungi and Fusarium oxysporum f. sp. niveum (FON) colonies, and increased soil enzyme activities in watermelon rhizosphere soil. Our results indicated that wheat and watermelon intercropping enhanced watermelon growth and decreased the incidence of Fusarium wilt in watermelon. These effects could be due to intercropping inducing physiological changes, regulating soil enzyme activities, and/or modulating soil microbial communities.

Studies on Fusarium wilt of Watermelon by using Antifungal Agents: An Invitro in Sights

Watermelon (Citrullus lanatus) is highly prone to Fusarium wilt, caused by a deadly soil-borne pathogen called Fusarium oxysporum f. sp. niveum (FON). Changes in production practices in crops, reduction in usage of fumigation with methyl bromide, crop rotation activities, and increasing human population, led to a resurgence of more virulent strains of FON. When resistant varieties are unavailable, the disease must be managed through a combination of integrated disease control practices. We conducted in vitro screening of botanicals where Allium cepa exhibited maximum inhibition of 80.32 and 82.62 percent at 5 and 10 percent concentrations, respectively. In vitro evaluation of bioagents revealed the maximum percent inhibition by T. harzianum (83.03%) followed by B. subtilis, A. flavus, P. fluorescence, T. viride, and A. niger. Fungicides by Poisoned Food Technique showed that a combination of carbendazim +mancozeb was effective in mycelial inhibition of 99.74 and 99.81 percent at 500 and 1000 ppm concentration respectively. The knowledge obtained from these in vitro findings can be applied in the field at a large scale for reducing and managing watermelon wilt disease.

Agronomic and disease responses of three watermelon (citrilus lanatus l.) varieties to fungicide spraying regimes in a tropical environment

usarium wilt disease is a major biotic factor limiting watermelon production in the derived savanna agroecology of southeast Nigeria. The study hypothesized that variety could interact with the fungicide spraying regime to improve the agronomic performance of watermelon (Citrulus lanatus (Thunb) Matsum and Nakai). The objectives of the study were: to assess the effect of fungicide spraying regimes on disease incidence, severity, and agronomic performance of three watermelon varieties; and to identify and characterize the fungal causal organism of Fusarium wilt of watermelon in the derived savanna agroecology of southeast Nigeria. Three varieties of watermelon (Jubaly, red diamond, and sweet sangaria) and four fungicide spraying regimes (No spray, weekly, bi-weekly, and tri-weekly spray) were evaluated for agronomic performance, and disease incidence and severity in a 3 × 4 factorial in a randomized complete block design replicated thrice. Data were collected on phenological, growth and yield indices, and disease incidence and severity. Analysis of variance was done using Genstat 16th edition and GraphPad Prism 9 was used to construct the graphs. Variety significantly affected agronomic traits as well as disease incidence and severity. The Jubaly variety performed better than other varieties in most of the growth and yield parameters measured despite recording higher disease incidence and severity scores. Weekly application of fungiforce recorded the lowest disease incidence (16.67, 16.67, and 50%) and severity (6.7, 6.7, and 10%) at 9, 10, and 11 weeks after planting, respectively. The interaction effect of the Jubaly variety and weekly spray was consistently higher in vine length, node number, fruit number, and fruit yield. Morphological isolation and identification of the fungal causal organism showed the presence of Fusarium species. Further molecular characterization by DNA extraction from the Fusarium isolate and sequencing using the Sanger sequencing method confirmed the presence of Fusarium equiseti. The basic local alignment search tool result showed 94–100% similarity with F. equiseti strain WZ-98 from the NCBI Gene bank. The organism was identified as F. equiseti based on the Phylogeny.

Antifungal profile and mechanism of bioinspired nanoscale magnesium against the agriculturally important pathogen Fusarium oxysporum f. sp. niveum

Biogenic MgNPs suppress watermelon Fusarium wilt, caused by Fusarium oxysporum f. sp. niveum (Fon), through precisly targeting peroxin biogenesis-centered signaling events in Fon.

Promotion of the symbiotic interaction between phagotrophic protists and beneficial bacteria, mediated via a balancing of soil nutrients, reduces the incidence of watermelon Fusarium wilt

Elucidating the microbial mechanisms that trigger Fusarium wilt represents a key step in addressing the barriers to sustainable cropping. However, from the perspective of the complete microbiome, the integrated role of soil nutrients and the microbial community in fields with different rate of wilt disease remains unclear. In this study, we examined the potential interrelationships among rhizosphere soil nutrients, bacteria, fungi, and protists in soils collected from fields in which watermelon had been cultivated for 8 years, and which were characterized by high (HW: 81.25%) or low (LW: 6.25%) incidence of watermelon Fusarium wilt. Soil from HW fields was found to contain a higher density of F. oxysporum (1.30-fold higher than that in the LW soil), along with higher contents of available phosphorus (1.31-fold higher), and available potassium (2.39-fold higher). In addition, the inter-kingdom correlation between protists and bacteria in the HW soil was 2.08-fold higher than that in LW soil. Furthermore, structural equation modeling revealed that an excess of soil available potassium enhances predation by potentially detrimental phagotrophic protists on potentially beneficial bacteria. Collectively, our findings indicate that a balanced nutrient input and the interaction between protists and beneficial bacteria play important roles in controlling the incidence of watermelon Fusarium wilt. These findings have implications for controlling the incidence of watermelon Fusarium wilt based on modifying the nutrient balance of soil and increasing the flora of certain beneficial protists (Cercomonas and Colpoda) that interact symbiotically with species of Bacillus.

Bacillus velezensis WB invokes soil suppression of Fusarium oxysporum f. sp. niveum by inducing particular taxa

Fusarium wilt of watermelon caused by Fusarium oxysporum f. sp. niveum (Fon) results in a substantial economic losses in agriculture. Biological control agents (BCAs) help in controlling the infection of Fon. However, it remains unclear whether the application of BCAs can invoke soil suppression by inducing specific microbial groups concerned with disease suppression to synergistically resist fungal pathogens. The objective of this project was to determine the microbial mechanisms involved in disease suppression by BCAs. Here, we conducted pot experiments to explore Fusarium wilt in watermelon and soil microbial communities in response to different treatments. We found that the application of Bacillus velezensis WB enhanced disease suppression in nonsterilized soil. We further found that disease suppression in the WF treatment was connected to influence on the resident soil microbial communities, explicitly by causing an increase in Hyphomicrobiaceae and Chitinophagaceae. Pathogen suppression by the above two families was further studied in laboratory and pot experiments. The results showed that a synthetic community consisting of specific bacterial taxa effectively decreased the incidence of Fusarium wilt. B. velezensis WB introduction impacts the resident soil microbiome and invokes soil suppression by inducing resident microbes. Our results should help in designing synthetic microbial communities to improve soil function.

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

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. 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 β-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. Our results indicated that B. velezensis WB is capable of inducing systemic resistance in watermelon against Fusarium wilt. © 2023 Society of Chemical Industry.

Antagonistic Activity of Streptomyces alfalfae 11F against Fusarium Wilt of Watermelon and Transcriptome Analysis Provides Insights into the Synthesis of Phenazine-1-Carboxamide.

Streptomyces alfalfa strain 11F has inhibitory effects on many phytopathogenic fungi and improves the establishment and biomass yield of switchgrass. However, the antagonistic effects of strain 11F on Fusarium wilt of watermelon and its secondary metabolites that contribute to its biocontrol activity are poorly understood. We evaluated the antagonistic and growth-promoting effects of strain 11F and conducted a transcriptome analysis to identify the metabolites contributing to antifungal activity. Strain 11F had marked inhibitory effects on six fungal pathogens. The incidence of Fusarium wilt of watermelon seedlings was decreased by 46.02%, while watermelon seedling growth was promoted, as indicated by plant height (8.7%), fresh weight (23.1%), and dry weight (60.0%). Clean RNA-sequencing data were annotated with 7553 functional genes. The 2582 differentially expressed genes (DEGs) detected in the Control vs. Case 2 comparison were divided into 42 subcategories of the biological process, cellular component, and molecular function Gene Ontology categories. Seven hundred and forty functional genes (55.47% of the DEGs) were assigned to Kyoto Encyclopedia of Genes and Genomes metabolic pathways, reflecting the complexity of the strain 11F metabolic regulatory system. The expression level of the gene phzF, which encodes an enzyme essential for phenazine-1-carboxylic acid (PCA) synthesis, was downregulated 3.7-fold between the 24 h and 48 h fermentation time points, suggesting that strain 11F can produce phenazine compounds. A phenazine compound from 11F was isolated and identified as phenazine-1-carboxamide (PCN), which contributed to the antagonistic activity against Fusarium oxysporum f. sp. niveum. PCA was speculated to be the synthetic precursor of PCN. The downregulation in phzF expression might be associated with the decrease in PCA accumulation and the increase in PCN synthesis in strain 11F from 24 to 48 h. Streptomyces alfalfae 11F protects watermelon seedlings from Fusarium wilt of watermelon and promotes seedling growth. The transcriptome analysis of strain 11F provides insights into the synthesis of PCN, which has antifungal activity against F. oxysporum f. sp. niveum of watermelon.

A Biocontrol Strain of Serratia plymuthica MM Promotes Growth and Controls Fusarium Wilt in Watermelon

Fusarium wilt, caused by Fusarium oxysporum f. sp. niveum (FON), is a predominant and devastating soil-borne disease that results in significant yield losses in watermelon cultivation. In this study, a strain MM isolated from the herbage rhizosphere soil, exhibited an inhibition rate of 65.46% against FON, leading to mycelial collapse, atrophy, and deformation. In pot experiments, strain MM effectively controlled Fusarium wilt of watermelon, showing a control efficacy of 74.07%. Through morphological observation and 16S rDNA gene sequencing, strain MM was identified as Serratia plymuthica. Additionally, S. plymuthica MM demonstrated antagonistic activity against eight plant pathogens, indicating that MM had broad-spectrum antifungal activity. The strain also exhibited the ability to synthesize siderophores and indole acetic acid (IAA), both of which are growth-promoting compounds. Moreover, strain MM secreted various extracellular enzymes, including protease, chitinase, β-glucanase, and cellulase. This ability allowed S. plymuthica MM to readily colonize watermelon roots and promote seedling growth. Inoculation with S. plymuthica MM increased the activity of PAL, POD, PPO, and CAT enzymes associated with watermelon defense. Furthermore, qRT-PCR analysis revealed up-regulation of LOX, POD, PAL, ClPR3, and C4H genes, which are related to plant disease resistance. The results indicated that S. plymuthica MM enhances watermelon plants’ resistance to FON by activating the JA, SA, and shikimic acid phenylpropanoid–lignin synthesis pathways. Gas chromatography–mass spectrometry (GC-MS) analysis of S. plymuthica MM culture supernatant identified piperazinedione, pyrrolo[1,2-a]pyrazine-1,4-dione, and octadecenamide as the main antimicrobial substances. Overall, S. plymuthica MM shows promise as a biocontrol agent against Fusarium wilt of watermelon, suggesting its potential for the development of a new biocontrol agent.

Control effect of root exudates from mycorrhizal watermelon seedlings on Fusarium wilt and the bacterial community in continuously cropped soil.

Watermelon (Citrullus lanatus) is susceptible to wilt disease caused by Fusarium oxysporum f. sp niveum (FON). AMF colonization alleviates watermelon wilt and regulates the composition of root exudates, but the effects of mycorrhizal watermelon root exudates on watermelon Fusarium wilt is not well understood. Root exudates of watermelon inoculated with AMF (Funeliformis mosseae or Glomus versiformme) were collected in this study. Then the root exudates of control plants and mycorrhizal plants were used to irrigate watermelon in continuous cropping soil, respectively. Meanwhile, the watermelon growth, antioxidant enzyme activity, rhizosphere soil enzyme activities and bacterial community composition, as well as the control effect on FON were analyzed. The results indicated that mycorrhizal watermelon root exudates promoted the growth of watermelon seedlings and increased soil enzyme activities, actinomyces, and the quantity of bacteria in rhizosphere soil. The proportion of Proteobacteria and Bacteroides was decreased, and the proportion of Actinobacteria, Firmicutes, and Chloroflexi in rhizosphere soil was increased when the seedlings were watered with high concentrations of mycorrhizal root exudates. The dominant bacterial genera in rhizosphere soil were Kaistobacter, Rhodanobacter, Thermomonas, Devosia, and Bacillus. The root exudates of mycorrhizal watermelon could reduce the disease index of Fusarium wilt by 6.7-30%, and five ml/L of watermelon root exudates inoculated with F. mosseae had the strongest inhibitory effect on watermelon Fusarium wilt. Our results suggest mycorrhizal watermelon root exudates changed the composition of bacteria and soil enzyme activities in rhizosphere soil, which increase the resistance of watermelon to Fusarium wilt and promoted the growth of plants in continuous cropping soil.