Plant-parasitic nematodes cause significant economic losses to a wide variety of crops. Chemical control is a widely used option for plantparasitic nematode management. However, chemical nematicides are now being reappraised in respect of environmental hazard, high costs, limited availability in many developing countries or their diminished effectiveness following repeated applications. This review presents progress made in the field of microbial antagonists of plant-parasitic nematodes, including nematophagous fungi, endophytic fungi, actinomycetes and bacteria. A wide variety of microorganisms are capable of repelling, inhibiting or killing plant-parasitic nematodes, but the commercialisation of these microorganisms lags far behind their resource investigation. One limiting factor is their inconsistent performance in the field. No matter how well suited a nematode antagonist is to a target nematode in a laboratory test, rational management decision can be made only by analysing the interactions naturally occurring among ''host plant-nematode target-soil-microbial control agent (MCA)-environment''. As we begin to develop a better understanding of the complex interactions, microbial control of nematodes will be more fine-tuned. Multidisciplinary collaboration and integration of biological control with other control methods will also contribute to more successful control practices.

BackgroundSubtilisin-like serine proteases play an important role in pathogenic fungi during the penetration and colonization of their hosts. In this study, we perform an evolutionary analysis of the subtilisin-like serine protease genes of subphylum Pezizomycotina to find if there are similar pathogenic mechanisms among the pathogenic fungi with different life styles, which utilize subtilisin-like serine proteases as virulence factors. Within Pezizomycotina, nematode-trapping fungi are unique because they capture soil nematodes using specialized trapping devices. Increasing evidence suggests subtilisin-like serine proteases from nematode-trapping fungi are involved in the penetration and digestion of nematode cuticles. Here we also conduct positive selection analysis on the subtilisin-like serine protease genes from nematode-trapping fungi.ResultsPhylogenetic analysis of 189 subtilisin-like serine protease genes from Pezizomycotina suggests five strongly-supported monophyletic clades. The subtilisin-like serine protease genes previously identified or presumed as endocellular proteases were clustered into one clade and diverged the earliest in the phylogeny. In addition, the cuticle-degrading protease genes from entomopathogenic and nematode-parasitic fungi were clustered together, indicating that they might have overlapping pathogenic mechanisms against insects and nematodes. Our experimental bioassays supported this conclusion. Interestingly, although they both function as cuticle-degrading proteases, the subtilisin-like serine protease genes from nematode-trapping fungi and nematode-parasitic fungi were not grouped together in the phylogenetic tree. Our evolutionary analysis revealed evidence for positive selection on the subtilisin-like serine protease genes of the nematode-trapping fungi.ConclusionsOur study provides new insights into the evolution of subtilisin-like serine protease genes in Pezizomycotina. Pezizomycotina subtilisins most likely evolved from endocellular to extracellular proteases. The entomopathogenic and nematode-parasitic fungi likely share similar properties in parasitism. In addition, our data provided better understanding about the duplications and subsequent functional divergence of subtilisin-like serine protease genes in Pezizomycotina. The evidence of positive selection detected in the subtilisin-like serine protease genes of nematode-trapping fungi in the present study suggests that the subtilisin-like serine proteases may have played important roles during the evolution of pathogenicity of nematode-trapping fungi against nematodes.

Aims:  To PCR‐amplify the full‐length genomic‐encoding sequence for one chitinase from the facultative fungal pathogen Paecilomyces lilacinus, analyse the DNA and deduced amino acid sequences and compare the amino acid sequence with chitinases reported from mycopathogens, entomopathogens and nematopathogens. Methods and Results:  The encoding gene (designated as PLC) was isolated using the degenerate PCR primers and the DNA‐Walking method. The gene is 1458 bp in length and contains three putative introns. A number of sequence motifs that might play a role in its regulation and function had also been found. Alignment of the translation product (designated as Plc, molecular mass of 45·783 kDa and pI of 5·65) with homologous sequences from other species showed that Plc belongs to Class V chitinase within the glycosyl hydrolase family 18. The phylogenetic and molecular evolutionary analysis using mega (Molecular Evolutionary Genetics Analysis) indicated that these chitinases from mycopathogens, entomopathogens and nematopathogens, the majority of which belong to glycosyl hydrolase family 18, were clustered into two well‐supported subgroups corresponding to ascomycetes fungal and nonfungal chitinases (bacteria, baculoviruses). Conclusions:  Our study showed that chitinases from mycoparasitic, entomopathogenic and nematophagous fungi are closely related to each other and reaffirmed the hypothesis that baculovirus chitinase is most likely to be of a bacterial origin – acquired by gene transfer. Bacterial and baculoviral chitinases in our study are potential pathogenicity factors; however, we still cannot ascribe any specific function to those chitinases from the fungi. Significance and Impact of the Study:  To our knowledge, this is the first report describing the chitinase gene and its translation product from Paecilomyces lilacinus, which constitutes the largest number of formulated biological nematicides reported so far, this is also the first study to analyse and resolve the phylogenetic and molecular evolutionary relationships among the chitinases produced by mycopathogens, entomopathogens and nematopathogens.

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