Plant parasitic nematodes represent a critical threat to global agriculture and ecosystems, and 28 biocontrol methods are becoming increasingly attractive as a means to combat nematode 29 infestation. Nematode-trapping fungi are a potentially useful biocontrol option, but further 30 research to enhance fungal pathogenicity will be needed before deployments are feasible. It is 31 known that nematode-trapping fungi can secrete cuticle-degrading serine proteases, which act 32 as key mediators of virulence against nematodes. Here, we describe the cloning and 33 characterization of the cuticle-degrading serine protease gene, AmSP1, from the 34 nematode-trapping fungus Arthrobotrys musiformis. Phylogenetic and selection force analysis 35 revealed a high degree of conservation of the AmSP1 catalytic and binding sites with 36 previously described serine proteases from other nematode-trapping fungi. The dN/dS ratio of 37 all six aligned-nematode-trapping fungi cuticle degrading serine proteases was less than 1, as 38 was the case of 386 individual codons, suggesting that the cuticle-degrading serine protease 39 gene has undergone purifying selection and is evolutionarily important for this group of fungi. 40
SummaryPolyporoid P hellinus fungi are ubiquitously present in the environment and play an important role in shaping forest ecology. Several species of P hellinus are notorious pathogens that can affect a broad variety of tree species in forest, plantation, orchard and urban habitats; however, current detection methods are overly complex and lack the sensitivity required to identify these pathogens at the species level in a timely fashion for effective infestation control. Here, we describe eight oligonucleotide microarray platforms for the simultaneous and specific detection of 17 important P hellinus species, using probes generated from the internal transcribed spacer regions unique to each species. The sensitivity, robustness and efficiency of this P hellinus microarray system was subsequently confirmed against template DNA from two key P hellinus species, as well as field samples collected from tree roots, trunks and surrounding soil. This system can provide early, specific and convenient detection of P hellinus species for forestry, arboriculture and quarantine inspection, and could potentially help to mitigate the environmental and economic impact of P hellinus‐related diseases.
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