Search Results (40)

A wide range of seed-borne and soil-borne plant pathogens belonging to various fungal and fungal-like species cause pre-emergence seed decay and post-emergence seedling blights of wheat and other small-grain cereal crops. To prevent the death of the seedlings, poor establishment and reduced stand of the crops, extensive crop rotations, planting good-quality seeds and seed treatments with fungicides are used on regular basis. This study is aimed at assessing the efficacy of pre-sowing seed treatments with cold atmospheric plasma for the disinfestation of winter wheat seed from economically important fungal and fungal-like pathogens. Uninoculated or surface-inoculated with Fusarium culmorum, Bipolaris sorokiniana or Pythium ultimum wheat seeds, the cultivar Madara was treated by cold plasma produced either by microwave torch (MW) or underwater diaphragm discharge (UW) with low power at very short treatment times, or remained untreated controls. As per the treatments, the seeds were sown in a ready-to-use growing medium comprising a mixture of light and dark moss peat (w:w) 90–95%, 5–10% perlite and 3–5 kg/m³ CaCO₃, having an electrical conductivity of 40 mS/m, pH (H₂O) of 5.5–6.5 and moisture content of 60–70%, filling in 250 × 250 × 70 mm aluminum flat seed trays (40 grains per tray, four trays per treatment). The plants were cultivated for 45 days in a growth chamber held at (20 ± 2) °C, set to a cycle of 8 h/night and 16 h/day under fluorescent light of 2000–3000 lux intensity. For each replicate, disease incidence (DI) was determined as the total percentage of missing, dead and apparently symptomatic plants. Seed treatment with a microwave plasma torch with a power of 16 W for 40 s significantly (p < 0.001) reduced seedling blights caused by F. culmorum, B. sorokiniana and P. ultimum by 46.8%, 51.0% and 77.3%, respectively, but limited the emergence of wheat seedlings by 15.9% on average. Simultaneously, the effectiveness of underwater discharge seed treatments reached an average of about a 60% reduction of seedling blight caused by F. culmorum and B. sorokiniana and about 37% of the disease caused by P. ultimum. Pre-sowing treatments with a MW plasma torch with an input power of 11 W and treatment time of 60, 90 or 120 s exposure also showed significant (p < 0.001) effects in controlling winter wheat seedling blights caused by the three pathogens. The effectiveness of the treatment increased with increasing the time period of exposure and reached full disease control (>80% reduction) for B. sorokiniana and P. ultimum seedling blights. This study demonstrated that pre-sowing treatment with a microwave plasma torch and underwater diaphragm discharge at a relatively low input power and short exposure time can be used for disinfestation and the effective control of seedling blights in winter wheat caused by seed-borne fungal pathogens, such as Fusarium culmorum and Bipolaris sorokiniana, and fungal-like oomycetes, such as Pythium ultimum. Full article

This study investigated whether viable cells, dead cells or cell-free supernatants (CFS) were responsible for the biocontrol effect of strains from two important bacterial genera, Pseudomonas and Lactobacillus, known for their antifungal properties against plant pathogens and food spoilage microorganisms. Specifically, the capability of these strains to produce extracellular hydrolytic enzymes on specified media was assessed, along with their effectiveness in inhibiting the mycelial growth of several phytopathogenic fungi (Fusarium oxysporum, Botrytis cinerea, Pythium ultimum and Rhizoctonia solani) using dual culture plate assays. Results from these inhibition assays revealed that P. fluorescens PF05 and L. plantarum LMG 23520 strains were the most effective in suppressing fungal growth, especially F. oxysporum. Therefore, further experiments were carried out to investigate the antifungal potential of the viable cells, heat-killed cells (HKC) and CFS from these strains against the germination of F. oxysporum spores. The viable cell trial proved successful, whereas HKC from the two bacterial isolates were ineffective against fungal spore germination. Conversely, the CFS of L. plantarum LMG 23520 was able to prevent fungal spore development for up to six days. The CFS of P. fluorescens PF05, instead, did not yield positive results. Additional studies are required to evaluate the potential inhibitory effects of the CFS from P. fluorescens PF05 and the HKC from both strains. Full article

To understand the molecular regulation of host defense responses in the pathosystem between apple roots and a necrotrophic oomycete pathogen Pythium ultimum, a series of transcriptome analyses have revealed a multi-phase and multi-layer defense tactic in apple root tissues. Among the most notable transcriptome changes during defense activation in apple roots, upregulation of genes involved in phenylpropanoid biosynthesis, transport of secondary metabolites, and lignin formation appeared to be the key defense themes which may crucially impact the outcome of plant–pathogen interactions. From our transcriptome datasets, the MdMATE52 gene, which encodes a MATE transporter, was shown to be differentially expressed between a resistant and a susceptible apple rootstock genotype in response to P. ultimum infection. The cis elements at promoter regions and sequence variations within coding regions of MdMATE52 were compared among several resistant and susceptible apple rootstock genotypes as well as various Malus species. The stronger upregulated expression patterns of MdMATE52 appeared to be correlated with the observed resistance traits among various genotypes. Our results suggested that minimal but clearly identifiable sequence variations may contribute to the genotype-specific expression and function of MdMATE52. The findings from this study should facilitate future experiments such as site-specific mutation and Crispr-based genome editing to define the regulation mechanisms of MdMATE52 and function during defense activation in apple roots. Full article

Lettuce is the most cultivated leafy vegetable in Greece; however, due to the adopted intensive cropping system, its cultivation is susceptible to many soilborne pathogens that cause significant yield and quality losses. In the current study, the impact of various soil disinfestation methods such as solarization, chemical disinfestation, and application of a biofungicide were evaluated in a commercial field that has been repeatedly used for lettuce cultivation. The populations of soilborne pathogens Rhizoctonia solani, Pythium ultimum, Fusarium oxysporum, and Fusarium equiseti were measured via qPCR before and after the implementation of the specific disinfestation methods. Although all the tested methods significantly reduced the population of the four soilborne pathogens, soil solarization was the most effective one. In addition, solarization reduced the number of lettuce plants affected by the pathogens R. solani and F. equiseti, and at the same time, significantly influenced the growth of lettuce plants. Amplicon sequence analysis of 16S rRNA-encoding genes used to study the soil bacterial community structure showed that Firmicutes, Proteobacteria, and Actinobacteria were the predominant bacterial phyla in soil samples. In general, solarization had positive effects on Firmicutes and negative effects on Proteobacteria and Actinobacteria; soil fumigation with dazomet increased the relative abundance of Firmicutes and Proteobacteria and reduced the corresponding values of Actinobacteria; and biofungicide had no significant effects on the three predominant bacterial phyla. The bacterial community composition and structure varied after the application of the soil disinfestation treatments since they imposed changes in the α- and β-diversity levels. The results of this study are expected to contribute towards implementing the most effective control method against the most common soilborne pathogens in intensively cultivated fields, such as those cultivated with leafy vegetables. Full article

Fungal diseases have always been a major problem for cantaloupe crops; however, synthetic fungicides are hazardous to humans and the environment. Consequently, a feasible alternative to fungicides without side effects could be by using bio agents and naturally occurring plants with antibacterial potential. This study has achieved a novel procedure for managing wilt and root rot diseases by potentially using Trichoderma sp. culture filtrates in consortium with plant extract of Calotropis procera, Rhizoctonia solani, Fusarium oxysporum, and Pythium ultimum, which were isolated from infected cantaloupe roots with identified root rot symptoms. The antagonistic activity of four Trichoderma isolates and analysis of antibiotics and filtrate enzymes of the most active Trichoderma isolate were determined as well as phytochemical analysis of C. procera plant extract using HPLC-UV. The obtained results showed that all Trichoderma isolates considerably lowered the radial growth of P. ultimum, R. solani, and F. oxysporum in varying degrees. The scanning electron micrographs illustrate the mycoparasitic nature of Trichoderma sp. on F. oxysporum. The phytochemical analysis of C. procera indicated that phenolic contents were the major compounds found in extracts, such as vanillin (46.79%), chlorogenic acid (30.24%), gallic acid (8.06%), and daidzein (3.45%) but including only a low amount of the flavonoid compounds rutin, naringenin, and hesperetin. The Pot experiment’s findings showed that cantaloupe was best protected against wilting and root rot diseases when it was treated with both Trichoderma sp. culture filtrates (10%) and C. procera extract of (15 mg/mL), both alone and in combination. This study demonstrates that the application of bio agent Trichoderma spp. filtrate with C. procera phenol extract appears useful for controlling wilting and root rot disease in cantaloupe. This innovative approach could be used as an alternative to chemical fungicide for the control of wilting and rot root diseases. Full article

Brewers’ spent grain (BSG) constitutes the primary by-product of the brewing industry. The valorization of BSG from a circular economy perspective is of high industrial interest. The objective of this study was the exploitation of BSG for the microbial production of branched-chain fatty acids (BCFAs) and polyunsaturated fatty acids (PUFAs), representing two different classes of high-value fatty acids (FAs). In the present study, this waste material underwent treatment with hot water in an autoclave and the resultant extract was utilized for the preparation of a novel liquid medium (BSG medium) to be employed for microbial fermentation. Screening and subsequent scaling-up experiments confirmed the suitability of the BSG medium to support the microbial production of various high-value FAs. In particular, Streptomyces jeddahensis and Conidiobolus heterosporus could be employed for BCFAs production, Pythium ultimum and Mortierella alpina could be used to provide cis-5,8,11,14,17-eicosapentaenoic acid (EPA) and arachidonic acid (ARA), whereas Mucor circinelloides, when grown in a BSG medium, was able to accumulate γ-linolenic acid (GLA). Full article

Currently, there are no standard management practices to counteract the adverse effects of fumigation on the soil microbiome. In this study, a variety of pre-plant soil amendments were examined for their ability to recruit and maintain apple rhizosphere microbiomes that are suppressive to pathogen re-infestation of fumigated orchard soils. The capacity of these amendments to improve other characteristics of soil productivity was also evaluated. Results suggest that composted chicken manure and liquid chitin are likely to be detrimental to plant and soil health when used as a post-fumigation soil amendment. In comparison, insect frass (IF) resulted in a significant increase in tree trunk diameter relative to the fumigated control. Following pathogen re-infestation of fumigated soil, however, IF induced a significant increase in Pythium ultimum in the rhizosphere. Therefore, IF can benefit the growth of young apple trees in fumigated soil but may stimulate pathogen activity upon re-infestation. To date, the possibility of using soil amendments to suppress pathogen re-infestation of fumigated soils has not been tested. Results from this study ground support the use of soil amendments as an intervention strategy for “steering” the soil and rhizosphere microbiome in more beneficial and/or prophylactic directions following fumigation. Full article

Pythium species are saprophytic or facultative plant pathogens that cause a variety of diseases. Usually, chemical anti-fungal seed dressing is applied in the conventional pre-harvest for seed protection. Nevertheless, recent legislative rules have created a ban on certain agrochemicals. Therefore, alternative eco-friendly methods have to be identified to ensure healthy field emergence and seedling development. In this study, a proof-of-concept was performed on the inactivation of Pythium ultimum Trow mycelia grown on potato dextrose broth agar (PBA) by plasma-processed air (PPA). Different plasma process parameters were applied using variation in gas flow of air through the microwave plasma generator and PPA exposure time. The PPA treatment was compared to the untreated and gas treated controls. The results showed a complete inactivation of P. ultimum mycelia after the PPA treatment. Inactivation efficiency was independent of the gas flow parameter and even shorter exposure times resulted in complete inactivation. To fully evaluate the potential of PPA as a possible seed hygiene measure, tests regarding the inactivation of P. ultimum after artificial inoculation onto seeds and/or studies using naturally infected seeds should be performed. This may be accompanied by monitoring the disease severity after the PPA treatment on a field scale. Full article

Pyrethrum (Tanacetum cinerariifolium) cultivation in Australia, which accounts for the majority of global production of natural insecticidal pyrethrins, is affected by a persistent yield decline which in part is caused by a complex of pathogens. Globisporangium and Pythium species were isolated from crown and roots of pyrethrum plants showing stunting and brown discoloration of crown tissue, and from soil adjacent to diseased plants from yield-decline-affected sites in Tasmania and Victoria, Australia. Ten known Globisporangium species (Globisporangium attrantheridium, G. erinaceum, G. intermedium, G. irregulare, G. macrosporum, G. recalcitrans, G. rostratifingens, G. sylvaticum, G. terrestris and G. ultimum var. ultimum), two new Globisporangium species (Globisporangium capense sp. nov. and Globisporangium commune sp. nov.) and three Pythium species (Pythium diclinum/lutarium, P. tracheiphilum and P. vanterpoolii) were identified through morphological studies and multigene phylogenetic analyses using ITS and Cox1 sequences. Globisporangium ultimum var. ultimum, G. sylvaticum, G. commune sp. nov. and G. irregulare were most abundant. Globisporangium attrantheridium, G. macrosporum and G. terrestris were reported for the first time in Australia. Seven Globisporangium species were pathogenic on both pyrethrum seeds (in vitro assays) and seedlings (glasshouse bioassays), while two Globisporangium species and three Pythium species only caused significant symptoms on pyrethrum seeds. Globisporangium irregulare and G. ultimum var. ultimum were the most aggressive species, causing pyrethrum seed rot, seedling damping-off and significant plant biomass reduction. This is the first report of Globisporangium and Pythium species causing disease in pyrethrum globally and suggests that oomycete species in the family Pythiaceae may have an important role in the yield decline of pyrethrum in Australia. Full article

Plant biostimulants are substances or microorganisms aimed at promoting plant growth by increasing the mineral nutrition efficiency, tolerance to environmental stress, and crop quality traits. This new category of crop inputs has been capturing the interest of both researchers and agriculture takeholders in light of the promising effects they could have on crop productivity and sustainability. This study investigated a variety of biostimulants for their effect on germination rates, plant health, chlorophyll fluorescence parameters, SPAD index, and growth of baby leaf lettuce and Batavia lettuce submitted to biotic (absence/presence of Pythium ultimum in the growing medium) or abiotic (0, 40, 80, and 120 mM NaCl L⁻¹ concentrations and −0.5, −2, −4, and −6 kPa water potentials) stresses when grown in a greenhouse under conventional and organic cultivation. The results obtained show that lettuce response to biostimulants was influenced by the type or level of stress applied and the growing system used. The effects of the tested biostimulants varied from strongly detrimental to strongly beneficial. Full article

Plant disease outbreaks are increasingly exacerbated by climate change and the conditions of stress combinations. They are negatively affecting crop yield and driving threats to food security in many areas of the world. Although synthetic pesticides offer relative success in the control of pests and plant diseases, they are often overused, and this method faces numerous drawbacks, including environmental toxicity, soil degradation, and adverse effects on human health. Therefore, alternatives are being developed and examined, including the biocontrol of pests and pathogens and biomass pyrolysis leading to wood vinegar that has shown great promise in agriculture and organic farming. However, while wood vinegar use is expanding and allows the control of numerous pests and bacterial and fungal diseases, its application to control oomycete diseases is limited. This study aimed to test wood vinegar for the control of oomycete plant pathogens from which six wood vinegars of pistachio, pomegranate, almond, pine, cypress, and walnut were produced. The inhibitory effects of volatile metabolites (semi-VOCs) of different wood vinegars concentrations (100%, 50%, 25%, 12.5%, and 6.25%) were examined against the hyphal growth of Globisporangium ultimum and Pythium aphanidermatum isolates. An in vitro analysis unambiguously demonstrated that for Globisporangium ultimum, the wood vinegar semi-VOCs of almond, pistachio (C 100% and 50%), and walnut (C 100%) totally inhibited mycelial growth. On the other hand, Pythium aphanidermatum, pistachio (C 100%, 50%, and 25%), and cypress (C 100%) expressed their abilities to completely inhibit the mycelial growth. Other treatments, including relevant concentrations of pine and pomegranate significantly inhibited the growth of mycelia of both species compared to the control (p ≤ 0.05). Therefore, wood vinegar could be considered a natural and organic product to use in agriculture to cope not only against pests, bacterial and fungal pests but also against emerging oomycete plant diseases. Full article

Oomycetes-borne diseases represent a serious problem for agriculture sustainability due to the high use of chemical products employed for their control. In recent years, increasing concerns on side effects associated with fungicide utilization have led to the reduction of the permissible modes of action, with the remaining ones continuously threatened by the increase of resistant strains in the pathogen populations. In this context, it is mandatory to develop new generation fungicides characterized by high specificity towards the target species and low environmental impact to guarantee the sustainability, productivity, and quality of food production. Fungicide discovery is a lengthy and costly process, and despite these urgent needs, poor description and formalization of high-throughput methodologies for screening the efficacy of active compounds are commonly reported for these kinds of organisms. In this study, a comprehensive picture of two high-throughput practices for efficient fungicide screening against plant-pathogenic oomycetes has been provided. Different protocols using multiwell plates were validated on approved crop protection products using Phytophthora infestans and Pythium ultimum as the model species. In addition, detailed statistical inputs useful for the analysis of data related to the efficacy of screenings are included. Full article

Some Trichoderma spp. exhibit natural abilities to reduce fungal diseases of plants through their mycoparasitic and antagonistic properties. In this study, we created new Trichoderma atroviride strains with elevated antifungal activity. This effect was achieved by improving the activity of cis-prenyltransferase, the main enzyme in dolichol synthesis, by expressing the RER2 gene from Saccharomyces cerevisiae. Since dolichyl phosphate is the carrier of carbohydrate residues during protein glycosylation, activation of its synthesis enhanced the activities of dolichyl-dependent enzymes, DPM synthase and N-acetylglucosamine transferase, as well as stimulated glycosylation of secretory proteins. Cellulases secreted by the transformants revealed significantly higher levels or activities compared to the control strain. Consequently, the resulting Trichoderma strains were more effective against the plant pathogens Pythium ultimum. Full article

The plant microbiome can be used to bolster plant defense against abiotic and biotic stresses. Some strains of endophytes, the microorganisms within plants, can directly inhibit the growth of plant fungal pathogens. A previously isolated endophyte from wild Populus (poplar), WPB of the species Burkholderia vietnamiensis, had robust in vitro antifungal activity against pathogen strains that are highly virulent and of concern to Pacific Northwest agriculture: Rhizoctonia solani AG-8, Fusarium culmorum 70110023, and Gaemannomyces graminis var. tritici (Ggt) ARS-A1, as well as activity against the oomycete, Pythium ultimum 217. A direct screening method was developed for isolation of additional anti-fungal endophytes from wild poplar extracts. By challenging pathogens directly with dilute extracts, eleven isolates were found to be inhibitory to at least two plant pathogen strains and were therefore chosen for further characterization. Genomic analysis was conducted to determine if these endophyte strains harbored genes known to be involved in antimicrobial activities. The newly isolated Bacillus strains had gene clusters for production of bacillomycin, fengicyn, and bacillibactin, while the gene cluster for the synthesis of sessilin, viscosin and tolaasin were found in the Pseudomonas strains. The biosynthesis gene cluster for occidiofungin (ocf) was present in the Burkholderia vietnamiensis WPB genome, and an ocf deletion mutant lost inhibitory activity against 3 of the 4 pathogens. The new isolates lacked the gene cluster for occidiofungin implying they employ different modes of action. Other symbiotic traits including nitrogen fixation, phosphate solubilization, and the production of auxins and siderophores were investigated. Although it will be necessary to conduct in vivo tests of the candidates with pathogen-infected agricultural crops, the wild poplar tree microbiome may be a rich source of beneficial endophyte strains with potential for biocontrol applications against a variety of pathogens and utilizing varying modes of action. Full article

The molecular mechanisms underpinning apple root resistance to infection by Pythium ultimum have not been carefully investigated until recently. A panoramic and high-resolution picture of genome-wide transcriptional networks regulating the defense activation in apple roots was obtained recently from a comprehensive transcriptome analysis. Among the most notable transcriptome changes are the upregulation of phenylpropanoid and flavonoid biosynthesis, transportation of secondary metabolites, and laccase-directed lignin formation. However, assessing cellular responses and anatomical changes in young roots of apple as a woody species of the Rosacea family remains challenging. Taking advantage of the autofluorescence of lignin and the specific staining by Wiesner reagent, the feasibility of using microscopy to detect lignin deposition in apple roots was evaluated. The preliminary results indicated that a combination of brightfield and fluorescence images may provide the opportunity to reveal the location and degree of lignification in sectioned roots. The procedure represents a proof-of-concept of using fluorescence microscopy to examine cellular features relevant to apple root resistance traits. The ability to detect subtle phenotypic variations, such as lignin deposition patterns, is critically needed to bridge the gap between genomic data and macro-level plant resistance phenotypes and to further identify the resistance mechanisms of apple root. Full article