The Dissemination of Metarhizium brunneum Conidia by Females of the Red Palm Weevil, Rhynchophorus ferrugineus, Suggests a New Mechanism for Prevention Practices.

Direct contact between the conidia of entomopathogenic fungi (EPF) and their host is a prerequisite to successful infection; the host can, therefore, be infected by both direct treatment and by transmission of fungal inoculum from infested surfaces. This unique characteristic makes EPF especially relevant for the control of cryptic insects. In the case of the red palm weevil (RPW) Rhynchophorus ferrugineus, the eggs and larvae are almost inaccessible to direct-contact treatment. The objective of the present study was to investigate the mechanism of conidia transmission from a treated surface to host eggs and larvae. Foam pieces infested with Metarhizium brunneum conidial powder, conidial suspension, or distilled water were used as a laying surface for RPW females. The number of eggs laid was not affected by the EPF treatments and ranged from 2 to 14 eggs per female. However, hatching rate and larval survival were significantly reduced in the conidial powder treatment, resulted in 1.5% hatching and no live larvae. In the conidial suspension treatment, 21% of laid eggs hatched, compared to 72% in the control treatment. In both M. brunneum treatments, females' proboscis, front legs and ovipositor were covered with conidia. The females transferred conidia in both treatments to the laying holes, reaching up to 15 mm in depth. This resulted in reduced egg-hatching rate and significant larval mortality due to fungal infection. The stronger effect on egg and larval survival using dry conidia seemed to result from better conidial adhesion to the female weevil in this formulation. In future studies, this dissemination mechanism will be examined as a prevention strategy in date plantations.

The SWC4 subunit of the SWR1 chromatin remodeling complex is involved in varying virulence of Metarhizium brunneum isolates offering role of epigenetic regulation of pathogenicity.

The host - pathogen interaction is a multifactorial process subject to a co-evolutionary arms race consisting of rapid changes in both host and pathogen, controlled at the genetic and epigenetic levels. Previously, we showed intra-species variation in disease progression and pathogenicity in aphids for Metarhizium brunneum isolates MbK and Mb7. Herein, we compared genomic, epigenetic, and metabolomic variations between these isolates and their effects on pathogenicity. Genomic variation could not completely explain the observed differences between the isolates. However, differential N6-adenine methylation (6 mA) and its correlation to reduced expression of the essential SWC4 subunit of SWR1 chromatin-remodelling complex (SWR1-C) led us to hypothesize a role for swc4 in the varying pathogenicity. Mutagenesis of the essential swc4 gene in MbKisolate resulted in reduction of secondary-metabolite (SM) secretion and impaired virulence in Galleria mellonella. Our results suggest the role of SWC4 in the regulation of SMs and the role of both SWC4 and SWR1-C in virulence of M. brunneum isolates. A better understanding of epigenetic regulation of SM production and secretion in entomopathogenic fungi may enable theirmanipulation for better biocontrol performance, and expand possibilities for environmentally friendly pest control.

Not Only a Formulation: The Effects of Pickering Emulsion on the Entomopathogenic Action of Metarhizium brunneum.

Growing global population and environmental concerns necessitate the transition from chemical to eco-friendly pest management. Entomopathogenic fungi (EPF) are rising candidates for this task due to their ease of growing, broad host range and unique disease process, allowing EPF to infect hosts directly through its cuticle. However, EPF's requirement for high humidity negates their integration into conventional agriculture. To mitigate this problem, we formulated Metarhizium brunneum conidia in an oil-in-water Pickering emulsion. Conidia in aqueous and emulsion formulations were sprayed on Ricinus communis leaves, and Spodoptera littoralis larvae were introduced under low or high humidity. The following were examined: conidial dispersion on leaf, larval mortality, conidial acquisition by larvae, effects on larval growth and feeding, and dynamic of disease progression. Emulsion was found to disperse conidia more efficiently and caused two-fold more adhesion of conidia to host cuticle. Mortality from conidia in emulsion was significantly higher than other treatments reaching 86.5% under high humidity. Emulsion was also found to significantly reduce larval growth and feeding, while conferring faster fungal growth in-host. Results suggest that a Pickering emulsion is able to improve physical interactions between the conidia and their surroundings, while weakening the host through a plethora of mechanisms, increasing the chance of an acute infection.