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Phylogenetic and phenotypic characterization of Fusarium oxysporum f. sp. niveum isolates from Florida-grown watermelon.

ABSTRACT: Fusarium wilt of watermelon (Citrullus lanatus) caused by Fusarium oxysporum f. sp. niveum (Fon), has become an increasing concern of farmers in the southeastern USA, especially in Florida. Management of this disease, most often through the use of resistant cultivars and crop rotation, requires an accurate understanding of an area's pathogen population structure and phenotypic characteristics. This study improved the understanding of the state's pathogen population by completing multilocus sequence analysis (MLSA) of two housekeeping genes (BT and TEF) and two loci (ITS and IGS), aggressiveness and race-determining bioassays on 72 isolates collected between 2011 and 2015 from major watermelon production areas in North, Central, and South Florida. Multilocus sequence analysis (MLSA) failed to group race 3 isolates into a single large clade; moreover, clade membership was not apparently correlated with aggressiveness (which varied both within and between clades), and only slightly with sampling location. The failure of multilocus sequence analysis using four highly conserved housekeeping genes and loci to clearly group and delineate known Fon races provides justification for future whole genome sequencing efforts whose more robust genomic comparisons will provide higher resolution of intra-species genetic distinctions. Consequently, these results suggest that identification of Fon isolates by race determination alone may fail to detect economically important phenotypic characteristics such as aggressiveness leading to inaccurate risk assessment.

SUBMITTER: Fulton JC

PROVIDER: S-EPMC7993609 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Genetic and phenotypic diversity of Fusarium oxysporum f. sp. niveum populations from watermelon in the southeastern United States.

Project description:Fusarium wilt of watermelon, caused by Fusarium oxysporum f. sp. niveum (FON), occurs worldwide and is responsible for substantial yield losses in watermelon-producing areas of the southeastern United States. Management of this disease largely relies on the use of integrated pest management (i.e., fungicides, resistant cultivars, crop rotation, etc.). Knowledge about race structure and genetic diversity of FON in the southeastern US is limited. To determine genetic diversity of the pathogen, FON isolates were collected from symptomatic watermelon plants in commercial fields in Georgia and Florida, USA, and identified based on morphological characteristics and PCR analysis using FON-specific primers. Discriminant analysis of principal components (DAPC) of 99 isolates genotyped with 15 simple sequence repeat (SSR) markers grouped the isolates in eight distinct clusters with two prominent clusters (clusters 1 and 8). Cluster 1 consisted of a total of 14 isolates, out of which 85.7% of the isolates were collected in Florida. However, most of the isolates (92.4%) in cluster 8 were collected in Georgia. Both DAPC and pairwise population differentiation analysis (ФPT) revealed that the genetic groups were closely associated with geographical locations of pathogen collection. Three races of FON (races 0, 2 and 3) were identified in the phenotypic analysis; with race 3 identified for the first time in Georgia. Overall, 5.1%, 38.9% and 55.9% of the isolates were identified as race 0, race 2 and race 3, respectively. The majority of the isolates in cluster 1 and cluster 8 belonged to either race 2 (35.6%) or race 3 (45.8%). Additionally, no relationship between genetic cluster assignment and races of the isolates was observed. The information obtained on genotypic and phenotypic diversity of FON in the southeastern US will help in development of effective disease management programs to combat Fusarium wilt.

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Transcriptional profiling of watermelon during its incompatible interaction with Fusarium oxysporum f. sp. Niveum

Project description:Watermelon (Citrullus lanatus) is one of the most important vegetable crops in the world and accounts for 20% of the world’s total area devoted to vegetable production. Fusarium wilt of watermelon is one of the most destructive diseases in watermelon worldwide. Transcriptome profiling of watermelon during its incompatible interactions with Fusarium oxysporum f.sp. niveum (FON) was generated using an Agilent custom microarray which contains 15,000 probes representing approximately 8,200 watermelon genes. A total of 24, 275, 596, 598, and 592 genes that are differentially expressed genes between FON- and mock-inoculated watermelon roots at 0.5, 1, 3, 5 and 8 days post inoculation (dpi), respectively, were identified. Bioinformatics analysis of these differentially expressed genes revealed that during the incompatible interaction between watermelon and FON, the expression of a number of pathogenesis-related (PR) genes, transcription factors, signaling/regulatory genes, and cell wall modification genes, was significantly induced. A number of genes for transporter proteins such as aquaporins were down-regulated, indicating that transporter proteins might contribute to the development of wilt symptoms after FON infection. In the incompatible interaction, most genes involved in biosynthesis of jasmonic acid (JA) showed expressed stronger and more sustained than those in compatible interaction in FON-infected tissues. Similarly, genes associated with shikimate-phenylpropanoid-lignin biosynthesis were also induced in incompatible interaction, but expression of these genes were not changed or repressed in the compatible interaction.

2011-06-09 | GSE29804 | GEO

Transcriptional profiling of watermelon during its incompatible interaction with Fusarium oxysporum f. sp. Niveum

2011-06-09 | E-GEOD-29804 | biostudies-arrayexpress

The FonSIX6 gene acts as an avirulence effector in the Fusarium oxysporum f. sp. niveum - watermelon pathosystem.

Project description:When infecting a host plant, the fungus Fusarium oxysporum secretes several effector proteins into the xylem tissue to promote virulence. However, in a host plant with an innate immune system involving analogous resistance proteins, the fungus effector proteins may trigger resistance, rather than promoting virulence. Identity of the effector genes of Fusarium oxysporum f. sp. niveum (Fon) races that affect watermelon (Citrullus lanatus) are currently unknown. In this study, the SIX6 (secreted in xylem protein 6) gene was identified in Fon races 0 and 1 but not in the more virulent Fon race 2. Disrupting the FonSIX6 gene in Fon race 1 did not affect the sporulation or growth rate of the fungus but significantly enhanced Fon virulence in watermelon, suggesting that the mutant ΔFon1SIX6 protein allowed evasion of R protein-mediated host resistance. Complementation of the wild-type race 2 (which lacks FonSIX6) with FonSIX6 reduced its virulence. These results provide evidence supporting the hypothesis that FonSIX6 is an avirulence gene. The identification of FonSix6 as an avirulence factor may be a first step in understanding the mechanisms of Fon virulence and resistance in watermelon and further elucidating the role of Six6 in Fusarium-plant interactions.

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The Transcriptomic Profile of Watermelon Is Affected by Zinc in the Presence of Fusarium oxysporum f. sp. niveum and Meloidogyne incognita.

Project description:Zinc (Zn) accumulation and deficiency affect plant response to pests and diseases differently in varying pathosystems. The concentrations of Zn in plants aid in priming defense signaling pathways and help in enhanced structural defenses against plant pathogens. Studies are lacking on how concentrations of Zn in watermelon plants influence defense against two important soil-borne pathogens: Fusarium oxysporum f. sp. niveum (FON) and southern root-knot nematode (RKN, Meloidogyne incognita). In this study a comparative transcriptomics evaluation of watermelon plants in response to high (1.2 ppm) and low (0.2 ppm) levels of Zn were determined. Differential transcript-level responses differed in watermelon plants when infected with FON or RKN or both under high- and low-Zn treatment regimes in a controlled hydroponics system. Higher numbers of differentially expressed genes (DEGs) were observed in high-Zn-treated than in low-Zn-treated non-inoculated plants, in plants inoculated with FON alone and in plants inoculated with RKN alone. However, in the co-inoculated system, low-Zn treatment had higher DEGs as compared to high-Zn treatment. In addition, most DEGs were significantly enriched in hormone signal transduction and MAPK signaling pathway, suggesting an induction of systemic resistance with high-Zn concentrations. Taken together, this study substantially expands transcriptome data resources and suggests a molecular potential framework for watermelon-Zn interaction in FON and RKN.

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Draft Genome Sequences of Three Fusarium oxysporum f. sp. niveum Isolates Used in Designing Markers for Race Differentiation.

Project description:Here, we report the draft genome sequences of three Fusarium oxysporum f. sp. niveum isolates that were used to design markers for molecular race differentiation. The isolates were collected from watermelon fields in Georgia (USA) and were determined to be different races of F. oxysporum f. sp. niveum using a traditional bioassay.

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Salicylic acid remodeling of the rhizosphere microbiome induces watermelon root resistance against Fusarium oxysporum f. sp. niveum infection.

Project description:Fusarium wilt disease poses a severe threat to watermelon cultivation by affecting the yield and quality of the fruit. We had previously found that the rhizosphere microbiome has a significant impact on the ability of watermelon plants to resist Fusarium wilt development and that salicylic acid (SA) is closely related to this phenomenon. Therefore, in this study, the role of SA as a mediator between plants and microbes in activating resistance against Fusarium oxysporum f. sp. niveum (FON) infection was explored through physiological, biochemical, and metagenomic sequencing experiments. We demonstrated that exogenous SA treatment could specifically increase some beneficial rhizosphere species that can confer resistance against FON inoculation, such as Rhodanobacter, Sphingomonas, and Micromonospora. Functional annotation analysis indicated that SA application significantly increased the relative abundance of glycoside hydrolase and polysaccharide lyase genes in the microbiome, which may play an essential role in increasing plant lipids. Moreover, network interaction analysis suggested that the highly expressed AAC6_IIC gene may be manipulated through SA signal transduction pathways. In conclusion, these results provide a novel strategy for controlling Fusarium wilt in watermelons from the perspective of environmental ecology, that is, by manipulating the rhizosphere microbiome through SA to control Fusarium wilt.

| S-EPMC9539938 | biostudies-literature

Antifungal Activity and Action Mode of Cuminic Acid from the Seeds of Cuminum cyminum L. against Fusarium oxysporum f. sp. Niveum (FON) Causing Fusarium Wilt on Watermelon.

Project description:In order to develop a novel biofungicide, the antifungal activity and action mode of cuminic acid from the seed of Cuminum cyminum L. against Fusarium oxysporum f. sp. niveum (FON) on watermelon was determined systematically. In this study, the median effective concentration (EC50) value for cuminic acid in inhibiting mycelial growth of FON was 22.53 μg/mL. After treatment with cuminic acid, the mycelial morphology was seriously influenced; cell membrane permeability and glycerol content were increased markedly, but pigment and mycotoxin (mainly fusaric acid) were significantly decreased. Synthesis genes of bikaverin (Bike1, Bike2 and Bike3) and fusaric acid (FUB1, FUB2, FUB3 and FUB4) both were downregulated compared with the control, as confirmed by quantitative RT-PCR. In greenhouse experiments, cuminic acid at all concentrations displayed significant bioactivities against FON. Importantly, significant enhancement of activities of SOD, POD, CAT and decrease of MDA content were observed after in vivo cuminic acid treatment on watermelon leaves. These indicated that cuminic acid not only showed high antifungal activity, but also could enhance the self-defense system of the host plant. Above all, cuminic acid showed the potential as a biofungicide to control FON.

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Project description:CCR4-Not complex is a multifunctional regulator that plays important roles in multiple cellular processes in eukaryotes. In the present study, the biological function of FonNot2, a core subunit of the CCR4-Not complex, was explored in Fusarium oxysporum f. sp. niveum (Fon), the causal agent of watermelon wilt disease. FonNot2 was expressed at higher levels in conidia and germinating conidia and during infection in Fon-inoculated watermelon roots than in mycelia. Targeted disruption of FonNot2 resulted in retarded vegetative growth, reduced conidia production, abnormal conidial morphology, and reduced virulence on watermelon. Scanning electron microscopy observation of infection behaviors and qRT-PCR analysis of in planta fungal growth revealed that the ΔFonNot2 mutant was defective in the ability to penetrate watermelon roots and showed reduced fungal biomass in root and stem of the inoculated plants. Phenotypic and biochemical analyses indicated that the ΔFonNot2 mutant displayed hypersensitivity to cell wall perturbing agents (e.g., Congo Red and Calcofluor White) and oxidative stress (e.g., H2O2 and paraquat), decreased fusaric acid content, and reduced reactive oxygen species (ROS) production during spore germination. Our data demonstrate that FonNot2 plays critical roles in regulating vegetable growth, conidiogenesis and conidia morphology, and virulence on watermelon via modulating cell wall integrity, oxidative stress response, ROS production and FA biosynthesis through the regulation of transcription of genes involved in multiple pathways.

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Inhibitory effect and mechanism of Tagetes erecta L. fungicide on Fusarium oxysporum f. sp. niveum.

Project description:Botanical fungicides comprise attractive alternatives to chemical fungicides because of their environmental compatibility. Flavonoids extracted from Tagetes erecta L. have an inhibitory effect on fusarium wilt in watermelons caused by Fusarium oxysporum f. sp. niveum (FON). In this study, we synthesized one of these flavonoids, 2,5-dicyclopentylidene cyclopentanone (Tagetes erecta L. fungicide (TEF)) and assessed its activity against FON. In vitro, TEF inhibited FON growth and killed FON cells directly. TEF also affected FON cell physiology and mycelial structure. In watermelon plants with fusarium wilt, TEF protected the leaf cell structure and improved the germination rate of infected seeds while increasing overall plant resistance. A TEF-resistant mutant (FONM) was created by chemical mutagenesis. FON and FONM were analysed using iTRAQ and RNA-Seq, which identified 422 differentially expressed proteins and 7817 differentially expressed mRNAs in the proteome and transcriptome, respectively. The FONM mutations caused changes in the cell membrane and cell wall, which may constitute the site of action of TEF. Together, these results demonstrate that TEF could effectively control the watermelon fusarium wilt caused by FON, possibly through the inhibition of sterol biosynthesis. The data presented here suggest that TEF represents a new potential botanical anti-fungal drug.

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