Publications

4493

Phytoplasmas classified in group 16SrXII infect a wide range of plants and are transmitted by polyphagous planthoppers of the family Cixiidae. Based on 16S rRNA gene sequence identity and biological properties, group 16SrXII encompasses several species, including ‘CandidatusPhytoplasma australiense’, ‘CandidatusPhytoplasma japonicum’ and ‘CandidatusPhytoplasma fragariae’. Other group 16SrXII phytoplasma strains are associated with stolbur disease in wild and cultivated herbaceous and woody plants and with bois noir disease in grapevines (Vitis viniferaL.). Such latter strains have been informally proposed to represent a separate species, ‘CandidatusPhytoplasma solani’, but a formal description of this taxon has not previously been published. In the present work, stolbur disease strain STOL11 (STOL) was distinguished from reference strains of previously described species of the ‘CandidatusPhytoplasma’ genus based on 16S rRNA gene sequence similarity and a unique signature sequence in the 16S rRNA gene. Other stolbur- and bois noir-associated (‘Ca.Phytoplasma solani’) strains shared >99 % 16S rRNA gene sequence similarity with strain STOL11 and contained the signature sequence. ‘Ca.Phytoplasma solani’ is the only phytoplasma known to be transmitted byHyalesthes obsoletus. Insect vectorship and molecular characteristics are consistent with the concept that diverse ‘Ca.Phytoplasma solani’ strains share common properties and represent an ecologically distinct gene pool. Phylogenetic analyses of 16S rRNA,tuf,secYandrplV–rpsCgene sequences supported this view and yielded congruent trees in which ‘Ca.Phytoplasma solani’ strains formed, within the group 16SrXII clade, a monophyletic subclade that was most closely related to, but distinct from, that of ‘Ca.Phytoplasma australiense’-related strains. Based on distinct molecular and biological properties, stolbur- and bois noir-associated strains are proposed to represent a novel species level taxon, ‘Ca.Phytoplasma solani’; STOL11 is designated the reference strain.

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 .

Summary Knowledge of intraspecific variation in symbioses may aid in understanding the ecology of widespread insects in different parts of their range. We investigated bacterial symbionts of A delges tsugae , a pest of hemlocks in eastern N orth A merica introduced from A sia. Amplification, cloning, and sequencing of bacterial 16 S rDNA , in situ hybridizations, and electron microscopy revealed that A . tsugae harbours up to five bacterial phylotypes, according to population. Three G ammaproteobacteria species are maternally transmitted. The first, designated ‘ C a . P seudomonas adelgestsugas’ resides in the haemocoel, and was detected in all populations except T aiwan. The second phylotype, ‘ C a . S erratia symbiotica’, resides in bacteriocytes of populations on T suga sieboldii in J apan and in E . N orth A merica. The third phylotype, designated ‘ C a . A nnandia adelgestsuga’, clustered within a lineage of several insect endosymbionts that included B uchnera aphidicola . It was detected in bacteriocytes in all populations, and in salivary glands of first instars. Two B etaproteobacteria phylotypes were detected in some J apanese T . sieboldii and eastern N orth A merica populations, and were observed only in salivary glands with no evidence of maternal transmission. Our results support the ideas that symbiont gain and loss has been volatile in adelgids, and that symbionts may help to trace the source of invasive species.