Dual-Transcriptomic, Microscopic, and Biocontrol Analyses of the Interaction Between the Bioeffector Pythium oligandrum and the Pythium Soft-Rot of Ginger Pathogen Pythium myriotylum

Biological control is a promising approach to suppress diseases caused by Pythium spp. such as Pythium soft rot of ginger caused by P. myriotylum. Unusually for a single genus, it also includes species that can antagonize Pythium plant pathogens, such as Pythium oligandrum. We investigated if a new isolate of P. oligandrum could antagonize P. myriotylum, what changes occurred in gene expression when P. oligandrum (antagonist) and P. myriotylum (host) interacted, and whether P. oligandrum could control soft-rot of ginger caused by P. myriotylum. An isolate of P. oligandrum, GAQ1, recovered from soil could antagonize P. myriotylum in a plate-based confrontation assay whereby P. myriotylum became non-viable. The loss of viability of P. myriotylum coupled with how P. oligandrum hyphae could coil around and penetrate the hyphae of P. myriotylum, indicated a predatory interaction. We investigated the transcriptional responses of P. myriotylum and P. oligandrum using dual-RNAseq at a stage in the confrontation where similar levels of total transcripts were measured from each species. As part of the transcriptional response of P. myriotylum to the presence of P. oligandrum, genes including a subset of putative Kazal-type protease inhibitors were strongly upregulated along with cellulases, elicitin-like proteins and genes involved in the repair of DNA double-strand breaks. In P. oligandrum, proteases, cellulases, and peroxidases featured prominently in the upregulated genes. The upregulation along with constitutive expression of P. oligandrum proteases appeared to be responded to by the upregulation of putative protease inhibitors from P. myriotylum, suggesting a P. myriotylum defensive strategy. Notwithstanding this P. myriotylum defensive strategy, P. oligandrum had a strong disease control effect on soft-rot of ginger caused by P. myriotylum. The newly isolated strain of P. oligandrum is a promising biocontrol agent for suppressing the soft-rot of ginger. The dual-RNAseq approach highlights responses of P. myriotylum that suggests features of a defensive strategy, and are perhaps another factor that may contribute to the variable success and durability of biological attempts to control diseases caused by Pythium spp.

A Comparison of the Ability of Some Commercially Produced Biological Control Agents to Protect Strawberry Plants against the Plant Pathogen Phytophthora cactorum

A comparison of the ability of commercially produced biological control agents—Contans, Gliorex, Hirundo, Polyversum, Prometheus, Clonoplus, Integral Pro and Xilon GR, completed with an isolate of Clonostachys rosea and of Pseudomonas sp.—to protect strawberry plants against Phytophthora cactorum was performed. The experiment was performed on strawberry cultivars Sonata, Karmen, and Wendy—cultivated in a cultivating room and greenhouse. The health of plants was affected negatively by the pathogen in all variants of biological agents used, but differences were seen in the rates of this decrease. The results revealed the ability of some tested agents to improve the growth of plants in the absence of the pathogen; the preparation Polyversum (Pythium oligandrum) was the most beneficial, in both the presence and absence of the pathogen. Contrarily, some agents alone decreased the health of plants; Integral Pro (Bacillus subtillis) and a strain of Pseudomonas sp. caused a deterioration in the health of the plants, even in the absence of a pathogen. The results of our analysis demonstrate the varied usefulness of all agents under unified environmental conditions; their effect seems to be dependent on the conditions and on the combination of the genotypes of all three participants in the interaction: plant–pathogen–antagonist.

Methods for suppressing Fusarium infection during malting and their effect on malt quality

The incidence of Fusarium head blight (FHB) in cereal grains such as barley and wheat is of growing concern due to climate change threatening food safety. Further processing of cereals by malting provides an ideal environment for the growth of Fusarium, leading to food safety concerns due to the production of mycotoxins, production challenges with the negative effects to malt and beer qualities, and economic loss owing to the field yield reduction. To improve food safety and product quality, different methods of fungal control have been investigated and reported in the literature. Traditional methods to control fungal growth and mycotoxin production have included chemical and physical methods, but these treatments led to worsened malt properties, limiting their applicability to the brewing industry. Biological control methods have, therefore, attracted wide interest as alternative treatments due to their ability to limit Fusarium growth and mycotoxin production in malting cereals without toxic by-products, thus exhibiting promise for improving food safety. Various biological agents have been investigated and applied in malting and have shown the potential to suppress Fusarium spp. growth and mycotoxin production. These agents include several lactic acid bacterial (LAB) species and Geotrichum candidum. Another promising biocontrol agent for malting control is Pythium oligandrum, which has successfully limited Fusarium infection in other agricultural crops. The review outlines the Fusarium-control methods reported referenced for the brewing industry and the present prospects in biological control applications on the promise of P. oligandrum as a novel agent for malting.

Effect of Seed Dressing and Soil Chemical Properties on Communities of Microorganisms Associated with Pre-Emergence Damping-Off of Broad Bean Seedlings

Combating soil pathogens that disable plant emergence is among the most difficult challenges of global agriculture. Legumes, preferred in sustainable cultivation systems, are particularly sensitive to pre-emergence damping-off of seedlings. Seed dressing is therefore a very important element in the cultivation technology. The aim of this study was to compare the impact of biological (Pythium oligandrum) and chemical (carboxin + thiuram) seed dressing on the quantitative and qualitative composition of microorganisms participating in the epidemiology of this disease, under specific hydrothermal conditions and chemical properties of the soil (pH, humus, macro-, and micronutrient). Microorganism identification was done using the MALDI-TOF MS (Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry) technique. Species were assigned to frequency groups, and populations of pathogens, saprophytes, and antagonists were identified. The biodiversity of these communities was expressed with Simpson’s Reciprocal, Shannon–Wiener, and Evenness (Shannon) indices. In individual variants of seed pre-treatment, the correlations between individual edaphic factors and the suppression of pre-emergence damping-off, the number of isolates obtained from infected seedlings, and the share of individual trophic groups of fungi were assessed. The main causes of pre-emergence damping-off of broad bean seedlings are Ilyonectria destructans, Globisporangium irregulare, Fusarium equiseti, Rhizoctonia solani, and Fusarium solani. Eliminating seed treatment results in a seedling mortality rate of 33.5%–42.5%. The effectiveness of the chemical protection product is 44.2% and 25.9%. Carboxin and thiuram reduce the diversity of microorganisms involved in the pathogenesis of pre-emergence damping-off and limit the presence of antagonistic fungi. Under the influence of P. oligandrum, there was a five-fold increase in the population of antagonists. An increase in humus in the soil reduces the percentage of diseased broad bean seedlings.

Novel approach of dermatophytosis eradication in shelters: effect of Pythium oligandrum on Microsporum canis in FIV or FeLV positive cats

Background Shelters and similar facilities with a high concentration and fluctuation of animals often have problems with various infections, which are usually difficult to solve in such environments and are very expensive to treat. This study investigated the eradication of Microsporum canis, the widespread cause of zoonotic dermatophytosis in shelters, even in immunosuppressed feline leukaemia virus or feline immunodeficiency virus positive cats. Results Our study showed the increased effectiveness of an alternative topical therapy for affected animals using the mycoparasitic fungus Pythium oligandrum, which is gentler and cheaper than the standard systemic treatment with itraconazole, and which can also be easily used as a preventative treatment. A decrease in the number of M. canis colonies was observed in cats treated with a preparation containing P. oligandrum 2 weeks after the start of therapy (2 cats with P-1 score, 2 cats with P-2 score, 5 cats with P-3 score) compared with the beginning of the study (9 cats with P-3 score = massive infection). The alternative topical therapy with a preparation containing P. oligandrum was significantly more effective compared with the commonly used systemic treatment using itraconazole 5 mg/kg in a 6-week pulse. After 16 weeks of application of the alternative topical therapy, the clinical signs of dermatophytosis were eliminated throughout the whole shelter. Conclusion The complete elimination of the clinical signs of dermatophytosis in all cats indicates that this therapy will be useful for the management and prevention of zoonotic dermatophytosis in animal shelters.

Type 2 Nep1-Like Proteins from the Biocontrol Oomycete Pythium oligandrum Suppress Phytophthora capsici Infection in Solanaceous Plants

As a non-pathogenic oomycete, the biocontrol agent Pythium oligandrum is able to control plant diseases through direct mycoparasite activity and boosting plant immune responses. Several P. oligandrum elicitors have been found to activate plant immunity as microbe-associated molecular patterns (MAMPs). Necrosis- and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) are a group of MAMPs widely distributed in eukaryotic and prokaryotic plant pathogens. However, little is known about their distribution and functions in P. oligandrum and its sister species Pythium periplocum. Here, we identified a total of 25 NLPs from P. oligandrum (PyolNLPs) and P. periplocum (PypeNLPs). Meanwhile, we found that PyolNLPs/PypeNLPs genes cluster in two chromosomal segments, and our analysis suggests that they expand by duplication and share a common origin totally different from that of pathogenic oomycetes. Nine PyolNLPs/PypeNLPs induced necrosis in Nicotiana benthamiana by agroinfiltration. Eight partially purified PyolNLPs/PypeNLPs were tested for their potential biocontrol activity. PyolNLP5 and PyolNLP7 showed necrosis-inducing activity in N. benthamiana via direct protein infiltration. At sufficient concentrations, they both significantly reduced disease severity and suppressed the in planta growth of Phytophthora capsici in solanaceous plants including N. benthamiana (tobacco), Solanum lycopersicum (tomato) and Capsicum annuum (pepper). Our assays suggest that the Phytophthora suppression effect of PyolNLP5 and PyolNLP7 is irrelevant to reactive oxygen species (ROS) accumulation. Instead, they induce the expression of antimicrobial plant defensin genes, and the induction depends on their conserved nlp24-like peptide pattern. This work demonstrates the biocontrol role of two P. oligandrum NLPs for solanaceous plants, which uncovers a novel approach of utilizing NLPs to develop bioactive formulae for oomycete pathogen control with no ROS-caused injury to plants.

Biocontrol Agents: Toolbox for the Screening of Weapons against Mycotoxigenic Fusarium

The aim of this study was to develop a set of experiments to screen and decipher the mechanisms of biocontrol agents (BCAs), isolated from commercial formulation, against two major mycotoxigenic fungi in cereals, Fusarium graminearum and Fusarium verticillioides. These two phytopathogens produce mycotoxins harmful to human and animal health and are responsible for the massive use of pesticides, for the protection of cereals. It is therefore essential to better understand the mechanisms of action of alternative control strategies such as the use of BCAs in order to optimize their applications. The early and late stages of interaction between BCAs and pathogens were investigated from germination of spores to the effects on perithecia (survival form of pathogen). The analysis of antagonist activities of BCAs revealed different strategies of biocontrol where chronological, process combination and specialization aspects of interactions are discussed. Streptomyces griseoviridis main strategy is based on antibiosis with the secretion of several compounds with anti-fungal and anti-germination activity, but also a mixture of hydrolytic enzymes to attack pathogens, which compensates for an important deficit in terms of spatial colonization capacity. It has good abilities in terms of nutritional competition. Trichoderma asperellum is capable of activating a very wide range of defenses and attacks combining the synthesis of various antifungal compounds (metabolite, enzymes, VOCs), with different targets (spores, mycelium, mycotoxins), and direct action by mycoparasitism and mycophagy. Concerning Pythium oligandrum, its efficiency is mainly due to its strong capacity to colonize the environment, with a direct action via microbial predation, stimulation of its reproduction at the contact of pathogens and the reduction of perithecia formation.

Myco-Suppression Analysis of Soybean (Glycine max) Damping-Off Caused by Pythium aphanidermatum

The role of Pythium oligandrum as a biocontrol agent against Pythium aphanidermatum was investigated to avoid the harmful impacts of fungicides. Three isolates of P. oligandrum (MS15, MS19, and MS31) were assessed facing the plant pathogenic P. aphanidermatum the causal agent of Glycine max damping-off. The tested Pythium species were recognized according to their cultural and microscopic characterizations. The identification was confirmed through sequencing of rDNA-ITS regions including the 5.8 S rDNA. The biocontrol agent, P. oligandrum, isolates decreased the mycelial growth of the pathogenic P. aphanidermatum with 71.3%, 67.1%, and 68.7% through mycoparasitism on CMA plates. While the half-strength millipore sterilized filtrates of P. oligandrum isolates degrade the pathogenic mycelial linear growth by 34.1%, 32.5%, and 31.7%, and reduce the mycelial dry weight of the pathogenic P. aphanidermatum by 40.1%, 37.4%, and 36.8%, respectively. Scanning electron microscopy (SEM) of the most effective antagonistic P. oligandrum isolate (MS15) interaction showed coiling, haustorial parts of P. oligandrum to P. aphanidermatum hyphae. Furthermore, P. oligandrum isolates were proven to enhance the germination of Glycine max seedling to 93.3% in damping-off infection using agar pots and promote germination of up to 80% during soil pot assay. On the other hand, P. oligandrum isolates increase the shoot, root lengths, and the number of lateral roots.