Search Results (18)

Verticillium longisporum, a soil-borne fungus responsible for Verticillium wilt, primarily colonizes members of the Brassicaceae family. Using Arabidopsis thaliana roots as an experimental host, we systematically identify V. longisporum-responsive genes and pathways through comprehensive transcriptomic analysis, alongside screening of potential hub genes and evaluation of infection-associated regulatory mechanisms. The GSE62537 dataset was retrieved from the Gene Expression Omnibus database. After performing GEO2R analysis and filtering out low-quality data, 222 differentially expressed genes (DEGs) were identified, of which 184 were upregulated. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed on these DEGs. A protein–protein interaction network was constructed using the STRING database. CytoHubba and CytoNCA plugins in Cytoscape v3.10.3 were used to analyze and evaluate this network; six hub genes and four functional gene modules were identified. The GeneMANIA database was used to construct a co-expression network for hub genes. Systematic screening of transcription factors within the 14 DEGs revealed the inclusion of the hub gene NAC042. Integrative bioinformatics analysis centered on NAC042 enabled prediction of a pathogen-responsive regulatory network architecture. We report V. longisporum-responsive components in Arabidopsis, providing insights for disease resistance studies in Brassicaceae crops. Full article

Verticillium stripe, a soilborne disease of canola (Brassica napus) caused by Verticillium longisporum, was first identified on the Canadian Prairies in 2014. Despite its increasing incidence, the impact of this disease on canola yields has not been quantified. To address this gap, the relationship between Verticillium stripe severity and yield was investigated in two canola hybrids, ‘45H31’ and ‘CS2000’, at two infested field sites near St. Albert, Alberta, in 2020 and 2021. In 2020, a year with above-average rainfall, both hybrids developed moderate levels of the disease, whereas in 2021, a drought year, symptoms and signs of infection were milder. Regression analysis indicated that seed yield per plant declined with increasing Verticillium stripe severity in both years of the study. In both hybrids, the relationship between disease severity and yield was best explained by second-degree quadratic equations. Although single-plant seed yield declined by up to 80% with increasing Verticillium stripe severity, these reductions did not translate into significant yield losses at the plot level, suggesting that losses experienced by individual plants were offset by reduced competition among the surviving plants. These results underscore the complexity of assessing disease impacts solely based on symptom severity. Full article

Verticillium stripe, caused by Verticillium longisporum, is an emerging disease of canola (Brassica napus) in Canada. Studies were conducted to assess the impact of pH on both the growth of V. longisporum and its virulence on the canola host. Fungal growth was assessed by measuring the colony diameter following 14 and 21 days of incubation on potato dextrose agar at varying pH levels (4.7, 5.5, 6.5, 7.4, or 8.6). The results indicated that colonies of V. longisporum were approximately 16% greater in diameter at pH 7.4 and 8.6 compared with those at pH 5.5. The impact of pH on disease development at the seedling stage was investigated using a semi-hydroponic system with different pH levels of 4.4, 5.4, 6.3, 7.5, and 8.4 in half-strength Hoagland’s solution. Verticillium stripe was most severe at pH 7.5 and 8.4 after a 10-day period in the semi-hydroponic system. In a second inoculation experiment, canola seedlings previously inoculated with the fungus were transplanted into potting mix amended to four pH levels (5.6, 6.4, 7.2, and 7.8). The transplants were cultivated under greenhouse conditions and evaluated for Verticillium stripe severity at plant maturity. Disease severity was greatest at pH 7.8. This is the first study on the effects of pH on V. longisporum in canola. It suggests a substantial risk of increased disease severity and yield losses due to Verticillium stripe in regions with neutral to slightly alkaline soils. Full article

Metarhizium brunneum is a soil-borne fungal entomopathogen that can be associated with plant roots. Previous studies have demonstrated that root colonization by beneficial fungi can directly affect soil-borne pathogens through competition and antibiosis and can activate a systemic response in plants, resulting in a primed state for a faster and/or stronger response to stressors. However, the mechanisms by which Metarhizium inoculation ameliorates symptoms caused by plant pathogens are not well known. This study evaluated the ability of M. brunneum to protect oilseed rape (Brassica napus L.) plants against the soil-borne pathogen Verticillium longisporum and investigated whether the observed effects are a result of direct interaction and/or plant-mediated effects. In vitro and greenhouse experiments were conducted to measure fungal colonization of the rhizosphere and plant tissues, and targeted gene expression analysis was used to evaluate the plant response. The results show that M. brunneum delayed pathogen colonization of plant root tissues, resulting in decreased disease symptoms. Direct competition and antibiosis were found to be part of the mechanisms, as M. brunneum growth was stimulated by the pathogen and inhibited the in vitro growth of V. longisporum. Additionally, M. brunneum changed the plant response to the pathogen by locally activating key defense hormones in the salicylic acid (SA) and abscisic acid (ABA) pathways. Using a split-root setup, it was demonstrated that there is a plant-mediated effect, as improved plant growth and decreased disease symptoms were observed when M. brunneum was in the systemic compartment. Moreover, a stronger systemic induction of the gene PR1 suggested a priming effect, involving the SA pathway. Overall, this study sheds light on the mechanisms underlying the protective effects of M. brunneum against soil-borne pathogens in oilseed rape plants, highlighting the potential of this fungal entomopathogen as a biocontrol agent in sustainable agriculture. Full article

Priming plants with beneficial microbes can establish rapid and robust resistance against numerous pathogens. Here, compelling evidence is provided that the treatment of rapeseed plants with Trichoderma harzianum OMG16 and Bacillus velezensis FZB42 induces defence activation against Verticillium longisporum infection. The relative expressions of the JA biosynthesis genes LOX2 and OPR3, the ET biosynthesis genes ACS2 and ACO4 and the SA biosynthesis and signalling genes ICS1 and PR1 were analysed separately in leaf, stem and root tissues using qRT-PCR. To successfully colonize rapeseed roots, the V. longisporum strain 43 pathogen suppressed the biosynthesis of JA, ET and SA hormones in non-primed plants. Priming led to fast and strong systemic responses of JA, ET and SA biosynthesis and signalling gene expression in each leaf, stem and root tissue. Moreover, the quantification of plant hormones via UHPLC-MS analysis revealed a 1.7- and 2.6-fold increase in endogenous JA and SA in shoots of primed plants, respectively. In roots, endogenous JA and SA levels increased up to 3.9- and 2.3-fold in Vl43-infected primed plants compared to non-primed plants, respectively. Taken together, these data indicate that microbial priming stimulates rapeseed defence responses against Verticillium infection and presumably transduces defence signals from the root to the upper parts of the plant via phytohormone signalling. Full article

The genus Verticillium is a group of ascomycete fungi that includes several pathogenic plant species. In 2011, a new taxonomic classification, proposed by Inderbitzin and coworkers (2011), re-defined the genus as Verticillium sensu stricto. The objective of our study was the re-classification of the fungal species held in the culture collection in the Slovenian Institute of Hop Research and Brewing in accordance with the newly established taxonomy. With the PCR marker system proposed by Inderbitzin and coworkers in 2011, we re-classified 88 Verticillium isolates out of the 105 samples that are held in the institute’s bank, which were obtained from different geographic locations in Europe, North America, and Japan, and from different host plants, including alfalfa, cotton, hop, olive, potato, and tomato. However, the PCR marker for the V. dahliae identification proved to be less specific, and it resulted in the positive amplification of Gibellulopsis nigrescens, V. isaacii, and V. longisporum. To enable the accurate distinction of the fungi, the SSR and LAMP markers were added to the analyses. The 12 newly identified SSR markers, which were used in simplex PCR reactions or in combination, enabled the accurate identification of all included Verticillium isolates and could potentially be used as biomarkers for rapid and easy species identification. Full article

Organic selenocyanates (RSeCN) are among the most reactive and biologically active Se species, often exhibiting a pronounced cytotoxic activity against mammalian cells and microorganisms. Various aromatic selenocyanates have been synthesized and, similar to some of the most Reactive Sulfur Species (RSS), such as allicin, found to be active against a range of bacteria, including Escherichia coli, Pseudomonas syringae and Micrococcus luteus, and fungi, including Verticillium dahlia, Verticillium longisporum, Alternaria brassicicola, and Botrytis cinerea, even via the gas phase. The highest antimicrobial activity has been observed for benzyl selenocyanate, which inhibited the growth of all bacteria considerably, even at the lowest tested concentration of 50 µM. Notably, neither the analogues thiocyanate (BTC) nor isothiocyanate (BITC) show any of these activities, rendering this selenium motif rather special in activity and mode of action. Eventually, these findings advocate a range of potential applications of organic selenocyanates in medicine and agriculture. Full article

Blackleg, caused by Leptosphaeria maculans, is an important disease of canola (Brassica napus). The pathogen can attack stems, leaves and pods, but basal stem cankers are most damaging and can result in significant yield losses. In Canada, Verticillium stripe (Verticillium longisporum) has recently emerged as another disease threat to canola. Symptoms of Verticillium stripe can resemble those of blackleg, and the two diseases may occur together. The effect of blackleg on yield was explored in field experiments with two canola hybrids and by evaluating a wider variety of hybrids in commercial crops in central Alberta, Canada. The impact on yield of L. maculans/V. longisporum interactions was also assessed under field and greenhouse conditions. In most hybrids, the relationship between blackleg severity and yield components was best explained by second-degree quadratic equations, although a linear relationship was found for one variety sampled in commercial fields. When L. maculans was co-inoculated with V. longisporum, blackleg severity and yield losses increased. In some cases, Verticillium stripe caused greater yield losses than blackleg. The results suggest that the interaction between L. maculans/V. longisporum may cause more severe losses in canola, highlighting the need for proactive disease management strategies. Full article

Polyketides are structurally diverse and physiologically active secondary metabolites produced by many organisms, including fungi. The biosynthesis of polyketides from acyl-CoA thioesters is catalyzed by polyketide synthases, PKSs. Polyketides play roles including in cell protection against oxidative stress, non-constitutive (toxic) roles in cell membranes, and promoting the survival of the host organisms. The genus Verticillium comprises many species that affect a wide range of organisms including plants, insects, and other fungi. Many are known as causal agents of Verticillium wilt diseases in plants. In this study, a comparative genomics approach involving several Verticillium species led us to evaluate the potential of Verticillium species for producing polyketides and to identify putative polyketide biosynthesis gene clusters. The next step was to characterize them and predict the types of polyketide compounds they might produce. We used publicly available sequences from ten species of Verticillium including V. dahliae, V. longisporum, V. nonalfalfae, V. alfalfae, V. nubilum, V. zaregamsianum, V. klebahnii, V. tricorpus, V. isaacii, and V. albo-atrum to identify and characterize PKS gene clusters by utilizing a range of bioinformatic and phylogenetic approaches. We found 32 putative PKS genes and possible clusters in the genomes of Verticillium species. All the clusters appear to be complete and functional. In addition, at least five clusters including putative DHN-melanin-, cytochalasin-, fusarielien-, fujikurin-, and lijiquinone-like compounds may belong to the active PKS repertoire of Verticillium. These results will pave the way for further functional studies to understand the role of these clusters. Full article

The production of volatile organic compounds (VOCs) represents a promising strategy of plant-beneficial bacteria to control soil-borne phytopathogens. Pseudomonas sp. PICF6 and Pseudomonas simiae PICF7 are two indigenous inhabitants of olive roots displaying effective biological control against Verticillium dahliae. Additionally, strain PICF7 is able to promote the growth of barley and Arabidopsis thaliana, VOCs being involved in the growth of the latter species. In this study, the antagonistic capacity of these endophytic bacteria against relevant phytopathogens (Verticillium spp., Rhizoctonia solani, Sclerotinia sclerotiorum and Fusarium oxysporum f.sp. lycopersici) was assessed. Under in vitro conditions, PICF6 and PICF7 were only able to antagonize representative isolates of V. dahliae and V. longisporum. Remarkably, both strains produced an impressive portfolio of up to twenty VOCs, that included compounds with reported antifungal (e.g., 1-undecene, (methyldisulfanyl) methane and 1-decene) or plant growth promoting (e.g., tridecane, 1-decene) activities. Moreover, their volatilomes differed strongly in the absence and presence of V. dahliae. For example, when co incubated with the defoliating pathotype of V. dahliae, the antifungal compound 4-methyl-2,6-bis(2-methyl-2-propanyl)phenol was produced. Results suggest that volatiles emitted by these endophytes may differ in their modes of action, and that potential benefits for the host needs further investigation in planta. Full article

Interaction of plants with the environment affects lipid metabolism. Changes in the pattern of phospholipids have been reported in response to abiotic stress, particularly accumulation of triacylglycerols, but less is known about the alteration of lipid metabolism in response to biotic stress and leaves have been more intensively studied than roots. This work investigates the levels of lipids in roots as well as leaves of Arabidopsis thaliana in response to pathogens and elicitor molecules by UPLC-TOF-MS. Triacylglycerol levels increased in roots and systemically in leaves upon treatment of roots with the fungus Verticillium longisporum. Upon spray infection of leaves with the bacterial pathogen Pseudomonas syringae, triacylglycerols accumulated locally in leaves but not in roots. Treatment of roots with a bacterial lipopolysaccharide elicitor induced a strong triacylglycerol accumulation in roots and leaves. Induction of the expression of the bacterial effector AVRRPM1 resulted in a dramatic increase of triacylglycerol levels in leaves, indicating that elicitor molecules are sufficient to induce accumulation of triacylglycerols. These results give insight into local and systemic changes to lipid metabolism in roots and leaves in response to biotic stresses. Full article

Verticillium wilt is one of the most important diseases of cauliflower and can lead to serious economic losses. In this study, two complementary strategies were explored to employ the antagonistic capacity of Verticillium isaacii towards Verticillium wilt of cauliflower. The first strategy focused on introducing V. isaacii Vt305 by artificial inoculation of cauliflower plantlets at the nursery stage. Two inoculum types (spores and microsclerotia of V. isaacii Vt305) and different concentrations of microsclerotia were tested in greenhouse and field trials. Seed treatment with 500 microsclerotia seed⁻¹ led to a satisfying biocontrol level of Verticillium wilt. In addition, the PHYTO-DRIP^® system was successful in delivering the microsclerotia to cauliflower seeds. The second strategy relied on the stimulation of the natural V. isaacii populations by rotating cauliflower with green manures and potato. Four green manure crops and potato were tested during multiple field experiments. Although these crops seemed to stimulate the V. isaacii soil population, this increase did not result in a control effect on Verticillium wilt of cauliflower in the short term. Importantly, our results indicate that the use of green manures is compatible with the application of V. isaacii Vt305 as biocontrol agent of Verticillium wilt in cauliflower. Full article

Reduction in atmospheric sulfur and intensified agriculture have led to sulfur deficiency, often correlated with a higher susceptibility to pathogens. The spread of fungal pathogens, such as the soil-born Verticillium longisporum, was observed. Defense responses of infected plants are linked to sulfur-containing compounds including glucosinolates (GSLs). Some pathogens infect their hosts at specific time periods during the day. To investigate the relation of sulfur-containing metabolites with diurnal effects of infection time points, Brassica napus plants cultivated at two different sulfur supplies, were infected with V. longisporum at four different time points during the day. It was demonstrated that 3, 7 and 14 days after inoculation the infected plants differed in their infection rate depending on the time point of infection. Additionally, infected plants had higher contents of sulfur-containing metabolites, such as specific GSLs, in comparison to non-infected plants. Sufficient sulfur fertilization was always reflected in higher contents of sulfur-containing compounds as well as a lower rate of infection compared to sulfur-deprived plants. On the microscopic level vascular occlusions in the hypocotyl were visible and the amount was dependent on the time point of infection. The results might be used to optimize sulfur fertilization to reduce susceptibility to V. longisporum. Full article

Microbiome management is a promising way to suppress verticillium wilt, a severe disease in Brassica caused by Verticillium longisporum. In order to improve current biocontrol strategies, we compared bacterial Verticillium antagonists in different assays using a hierarchical selection and evaluation scheme, and we integrated outcomes of our previous studies. The result was strongly dependent on the assessment method chosen (in vitro, in vivo, in situ), on the growth conditions of the plants and their genotype. The most promising biocontrol candidate identified was a Brassica endophyte Serratia plymuthica F20. Positive results were confirmed in field trials and by microscopically visualizing the three-way interaction. Applying antagonists in seed treatment contributes to an exceptionally low ecological footprint, supporting efficient economic and ecological solutions to controlling verticillium wilt. Indigenous microbiome, especially soil and seed microbiome, has been identified as key to understanding disease outbreaks and suppression. We suggest that verticillium wilt is a microbiome-driven disease caused by a reduction in microbial diversity within seeds and in the soil surrounding them. We strongly recommend integrating microbiome data in the development of new biocontrol and breeding strategies and combining both strategies with the aim of designing healthy microbiomes, thus making plants more resilient toward soil-borne pathogens. Full article

Verticillium stripe in canola (Brassica napus L.) caused by Verticillium longisporum was first reported in Manitoba in 2014. In this study, Brassica crops including canola, mustard (Brassica juncea) and radish (Raphanus sativus) with visible symptoms of Verticillium stripe were collected from Portage La Prairie, Manitoba, and the pathogens were isolated. Isolates from canola and radish were identified to V. longisporum, which produced longer conidia (7.92–12.00 µm) than Verticillium dahliae (4.32–7.04 µm). An isolate derived from mustard was characterized as V. dahliae. Molecular diagnostics with 18S rDNA, 5.8S rDNA and mating-type marker primers were used to confirm the identification of Verticillium isolates. PCR-RFLP of the mitochondrial small subunit rDNA and the cytochrome b gene were also employed to distinguish V. longisporum isolates from V. dahliae. The multi-gene characterization approach allowed for lineage determination, and V. longisporum isolates from canola and radish were in the A1/D1 group. Isolates of Verticillium longisporum from canola inoculated onto the canola cultivar ‘Westar’ caused symptoms of stem striping, stunting and short plants. Re-isolated fungal strains from infected stems were again inoculated onto canola plants, in order to confirm that V. longisporum was the causal agent of Verticillium stripe disease in the pathogenicity test. Full article