The entomopathogenic fungus Lecanicillium muscarium (Petch) Zare and Gams is currently being developed as a biocontrol agent against insect pests, as well as some plant-pathogenic fungi and bacteria. Data about its activity against plant-parasitic nematodes exist, but are relatively limited. To expand this understanding, we investigated the biocontrol efficiency of three isolates of L. muscarium (Lm) against the root knot nematode, Meloidogyne incognita, in both in vitro and in vivo conditions. In our experiments, the maximum number of nematode eggs, juveniles (J2s), females, and egg masses that were parasitized were quantified after a 72-h exposure to the fungus. The isolate Lm1 was designated as the best biocontrol agent against nematode eggs as well as J2s. It showed the highest colonization of eggs and significantly decreased egg hatching events. The results from two additional isolates, Lm2 and Lm3, were also significant (P = 0.05) but less pronounced than those observed with Lm1. L. muscarium treatments had significant (P = 0.05) positive effects on plant shoot and root growth compared with the growth of control plants. These results suggest the effectiveness of the fungus may be due to either the infection of eggs and J2s, or the production of secondary metabolites that induced plant defense mechanisms and lead to systemic resistance. Our study demonstrates that L. muscarium could be used as a potential biocontrol agent against root knot nematodes.

Caenorhabditis elegans presents functioning, biologically relevant phenotypes and is frequently used as a bioindicator of toxicity. However, most C. elegans in vivo effect-assessment methods are laborious and time consuming. Therefore, we developed a novel method to measure the oxygen consumption rate of C. elegans as a sublethal endpoint of toxicity. This protocol was tested by exposing 50 larval stage one C. elegans individuals for 48 h (at 20 °C) to different concentrations of two toxicants i.e. benzylcetyldimethylammonium chloride (BAC-C16) and cadmium (Cd). Following exposures, the oxygen consumption rate of the C. elegans individuals were measured using the high-throughput functionality of the Seahorse XF^e96 Extracellular Flux Analyzer. Dose-response curves for BAC-C16 (R² = 0.93; P = 0.001) and Cd (R² = 0.98; P = 0.001) were created. Furthermore, a strong, positive correlation was evidenced between C. elegans oxygen consumption rate and a commonly used, ecologically relevant endpoint of toxicity (growth inhibition) for BAC-C16 (R² = 0.93; P = 0.0001) and Cd (R² = 0.91; P = 0.0001). The data presented in this study show that C. elegans oxygen consumption rate can be used as a promising functional measurement of toxicity.

Clubroot, caused by Plasmodiophora brassicae, is a crucial oilseed rape disease worldwide. Information on the virulence of P. brassicae populations is essential to apply disease control with proper clubroot-resistant cultivars. In 2016–2020, 84 isolates of P. brassicae were collected in the Czech Republic (CZ), Germany (DE), Poland (PL), and Sweden (SW). Pathotypes were designated using 17 Brassica hosts, including the European Clubroot Differentials (ECD), Somé set, and clubroot-resistant oilseed rape cv. Mendel. According to the ECD set, virulence analyses differentiated the isolates into 42 pathotypes. The most common pathotypes were 16/31/31 (in DE, PL, and SW) and 16/06/12 (in CZ, DE, and PL). Six pathotypes were found according to the Somé set, including 1–4 pathotypes per country. P1 was most prevalent in DE, PL, and SW, while P3 was abundant in CZ, DE, and PL. The current study provides clear evidence for a shift towards increased virulence in P. brassicae populations compared to previous studies. Several isolates overcame the resistance of cv. Mendel and of Brassica rapa genotypes ECD 01 to ECD 04. Considering all investigated samples, significant negative correlations were found between clubroot incidence and the frequency of oilseed rape in crop rotation, as for clubroot incidence and soil pH.