Burckhardt, Daniel

In Graneros, O’Higgins Region, Chile, the mallow psyllid (Russelliana solanicola Tuthill, 1959) from Malva nicaeensis L. was identified as a potential vector of ‘Candidatus Phytoplasma pruni’. Over an 8-month period, 2089 specimens of a species of Psylloidea, including immatures and adults, were captured. We only selected the adults used for transmission trials in Catharanthus roseus (L.) G. Don (periwinkle) plants. By nested PCR, using primer pairs for phytoplasma detection in 16S rRNA and IdpA genes, 7 out of 113 (6.2%) periwinkle plants used in transmission trials were found to be infected by phytoplasmas. Insects that fed on these plants also tested positive for the same phytoplasmas. Periwinkle plants never showed virescence and phyllody, as commonly occurs with phytoplasma 16SrIII-J infection due to the effector SAP54. In this case, using primer pairs for the SAP54 gene, an amplification product was never obtained. Virtual restriction fragment length polymorphism (RFLP) analysis of F2nR2 fragments indicated that the phytoplasma, found in both periwinkle plants and insects used in transmission trials, belongs to the 16SrIII-J ribosomal subgroup. The COI gene of the psyllids samples was amplified and sequenced, showing a similarity ranging from 84.84% to 85.02% with R. solanicola from Solanum tuberosum L. The mitochondrial genome of the psyllid was also sequenced, revealing a 14,835 bp circular DNA molecule with 37 genes. The mallow psyllid transmitted the phytoplasma 16SrIII-J to periwinkle plants. The molecular identification of the insect does not match the morphological one, indicating that the mallow psyllid may constitute a cryptic species within the polyphagous R. solanicola species. This is the first report of a psyllid as a vector of the phytoplasma 16SrIII-J.

Summary Many hemipterans are associated with symbiotic bacteria, which are usually found intracellularly in specific bacteriomes. In this study, we provide the first molecular identification of the bacteriome‐associated, obligate endosymbiont in a G ondwanan relict insect taxon, the moss bugs ( H emiptera: C oleorrhyncha: P eloridiidae), which represents one of the oldest lineages within the H emiptera. Endosymbiotic associations of fifteen species of the family were analysed, covering representatives from S outh A merica, A ustralia/ T asmania and N ew Z ealand. Phylogenetic analysis based on four kilobases of 16 S –23 S r RNA gene fragments showed that the obligate endosymbiont of P eloridiidae constitute a so far unknown group of G ammaproteobacteria which is named here ‘ Candidatus Evansia muelleri’. They are related to the sternorrhynchous endosymbionts C andidatus   P ortiera and C andidatus   C arsonella. Comparison of the primary‐endosymbiont and host ( COI  + 28 S rRNA ) trees showed overall congruence indicating co‐speciation the hosts and their symbionts. The distribution of the endosymbiont within the insect body and its transmission was studied using FISH . The endosymbionts were detected endocellularly in a pair of bacteriomes as well as in the ‘symbiont ball’ of the posterior pole of each developing oocyte. Furthermore, ultrastructural analysis of the M alpighian tubules revealed that most host nuclei are infected by an endosymbiotic, intranuclear bacterium that was determined as an A lphaproteobacterium of the genus R ickettsia .