Essential Role of WetA, but No Role of VosA, in Asexual Development, Conidial Maturation and Insect Pathogenicity of Metarhizium robertsii

WetA and VosA regulate conidiation and conidial maturation required for the life cycle of Beauveria bassiana, like they do in Aspergillus, but remain functionally unexplored in Metarhizium robertsii, another hypocrealean pathogen considered to have evolved insect pathogenicity ~130 million years later than B. bassiana. This study reveals a similar role of WetA ortholog in asexual development, conidial maturation, and insect pathogenicity, and also its distinctive role in mediating some other conidial maturation-related cellular events, but has functional redundancy of VosA in M. robertsii. ABSTRACT Conidial maturation, which is crucial for conidial quality, is controlled by the asexual development activator WetA and the downstream, velvety protein VosA in Aspergillus. Their orthologs have proved functional in conidial quality control of Beauveria bassiana, as seen in Aspergillus, but are functionally unexplored, in Metarhizium robertsii, another hypocrealean insect pathogen. Here, WetA and VosA prove essential and nonessential for M. robertsii's life cycle, respectively. Disruption of wetA increased hyphal sensitivity to oxidative stress and Congo red-induced cell wall stress, but had little impact on radial growth. The ΔwetA mutant was severely compromised in conidiation capacity and conidial quality, which was featured by slower germination, decreased UV resistance, reduced hydrophobicity, and deformed hydrophobin rodlet bundles that were assembled onto conidial coat. The mutant's virulence was greatly attenuated via normal infection due to a blockage of infection-required cellular processes. All examined phenotypes were unaffected for the ΔvosA mutant. Intriguingly, mannitol was much less accumulated in the 7- and 15-day-old cultures of ΔwetA and ΔvosA than of control strains, while accumulated trehalose was not detectable at all, revealing little a link of intracellular polyol accumulation to conidial maturation. Transcriptomic analysis revealed differential regulation of 160 genes (up/down ratio: 72:88) in ΔwetA. These genes were mostly involved in cellular component, biological process, and molecular function but rarely associated with asexual development. Conclusively, WetA plays a relatively conserved role in M. robertsii’s spore surface structure, and also a differentiated role in some other cellular processes associated with conidial maturation. VosA is functionally redundant in M. robertsii unlike its ortholog in B. bassiana. IMPORTANCE WetA and VosA regulate conidiation and conidial maturation required for the life cycle of Beauveria bassiana, like they do in Aspergillus, but remain functionally unexplored in Metarhizium robertsii, another hypocrealean pathogen considered to have evolved insect pathogenicity ~130 million years later than B. bassiana. This study reveals a similar role of WetA ortholog in asexual development, conidial maturation, and insect pathogenicity, and also its distinctive role in mediating some other conidial maturation-related cellular events, but has functional redundancy of VosA in M. robertsii. The maturation process vital for conidial quality proves dependent on a role of WetA in spore wall assembly but is independent of its role in intracellular polyol accumulation. Transcriptomic analysis reveals a link of WetA to 160 genes involved in cellular component, biological process, and molecular function. Our study unveils that M. robertsii WetA or VosA is functionally differential or different from those learned in B. bassiana and other ascomycetes.