Christina Schuster

Genetic and Electron-Microscopic Characterization of ‘Rickettsiella agriotidis’, a new Rickettsiella Pathotype Associated with Wireworm, Agriotes sp. (Coleoptera: Elateridae)

Wireworms, the polyphagous larvae of click beetles belonging to the genus Agriotes (Coleoptera: Elateridae), are severe and widespread agricultural pests affecting numerous crops. A previously unknown intracellular bacterium has been identified in a diseased Agriotes larva. Microscopic studies revealed the subcellular structures characteristic of Rickettsiella infections. Molecular phylogenetic analysis based on 16S ribosomal RNA and signal recognition particle receptor (FtsY) encoding sequences demonstrates that the wireworm pathogen belongs to the taxonomic genus Rickettsiella. Therefore, the new pathotype designation ‘R. agriotidis’ is proposed to refer to this organism. Moreover, genetic analysis makes it likely that—on the basis of the currently accepted organization of the genus Rickettsiella—this new pathotype should be considered a synonym of the nomenclatural type species, Rickettsiella popilliae.

Multilocus sequence analysis (MLSA) of ‘Rickettsiella costelytrae' and ‘Rickettsiella pyronotae’, intracellular bacterial entomopathogens from New Zealand

Larvae of scarab beetles live in the soil and are frequently hosts for microbial pathogens. In New Zealand, larvae of the grass grub, Costelytrae zealandica (Coleoptera: Scarabaeidae), and manuka beetles, Pyronota spp. (Coleoptera: Scarabaeidae), have been collected from field populations showing loss of vigour and a whitened appearance. Diagnosis indicated an intracellular infection of fat body tissues by Rickettsiella‐like micro‐organisms. Rickettsiella bacteria are under evaluation as a possible new source of insect bio‐control agents for important agricultural pests as, e.g. scarabaeid and elaterid larvae. The present study aimed at the unequivocal molecular taxonomic identification and comparison of the bacteria associated with Costelytra and Pyronota.

Ribosomal RNA phylogeny of bacterial and fungal pathogens of Agriotes wireworms

Wireworms, the polyphagous larvae of click beetles belonging to the genus Agriotes (Coleoptera: Elateridae), are severe and widespread agricultural pests affecting numerous crops. Biological control agents and methods for this general pest are highly solicited. In a screening for microbial Agriotes pathogens, an intracellular bacterium and a mitosporic fungus were isolated. Phylogenetic analysis based on ribosomal RNA operon sequences of both micro-organisms corroborated their previous morphology-based taxonomic classification. The bacterial pathogen has been assigned to the taxonomic genus Rickettsiella (Gammaproteobacteria) wherein it represents a new pathotype, ‘Rickettsiella agriotidis’, that appears most closely related to subjective synonyms of the nomenclatural type species, Rickettsiella popillae. The fungal pathogen has been shown to belong to the form-species Beauveria bassiana, i.e., an obligate anamorph related to the genus Cordyceps (Ascomycota: Hypocreales). Furthermore, the B. bassiana strain from Agriotes has been shown to be potentially susceptible to identification by gIi-diagnosis, i.e., a diagnostic method making use of the strain-specific presence of self-splicing group-I introns within the ribosomal RNA operons of certain hyphomycetous fungi.

Multilocus Sequence Analysis (MLSA) of ‘Rickettsiella agriotidis’, an Intracellular Bacterial Pathogen of Agriotes Wireworms

Wireworms, the polyphagous larvae of click beetles belonging to the genus Agriotes (Coleoptera: Elateridae) are severe and widespread agricultural pests that affect numerous crops globally. A new bacterial specimen identified in diseased wireworms had previously been shown by microscopy and 16S ribosomal RNA (rRNA) gene-based phylogenetic reconstruction to belong to the taxonomic genus Rickettsiella (Gammaproteobacteria) that comprises intracellular bacteria associated with and typically pathogenic for a wide range of arthropods. Going beyond these earlier results obtained from rRNA phylogenies, multilocus sequence analysis (MLSA) using a four marker scheme has been employed in the molecular taxonomic characterization of the new Rickettsiella pathotype, referred to as ‘Rickettsiella agriotidis’. In combination with likelihood-based significance testing, the MLSA approach demonstrated the close phylogenetic relationship of ‘R. agriotidis’ to the pathotypes ‘Rickettsiella melolonthae’ and ‘Rickettsiella tipulae’, i.e., subjective synonyms of the nomenclatural type species, Rickettsiella popilliae. ‘R. agriotidis’ forms, therefore, part of a Rickettsiella pathotype complex that most likely represents the species R. popilliae. As there are currently no genetic data available from the R. popilliae type strain, the respective assignment cannot be corroborated directly. However, an alternative taxonomic assignment to the species Rickettsiella grylli has been positively ruled out by significance testing. MLSA has been shown to provide a more powerful tool for taxonomic delineation within the genus Rickettsiella as compared to 16S rRNA phylogenetics. However, the limitations of the present MLSA scheme for the sub-species level classification of ‘R. agriotidis’ and further R. popilliae synonyms has been critically evaluated.