Division of labor within psyllids: metagenomics reveals an ancient dual endosymbiosis with metabolic complementarity in the genus Cacopsylla.

IMPORTANCE: Heritable beneficial bacterial endosymbionts have been crucial for the evolutionary success of numerous insects by enabling the exploitation of nutritionally limited food sources. Herein, we describe a previously unknown dual endosymbiosis in the psyllid genus Cacopsylla, consisting of the primary endosymbiont "Candidatus Carsonella ruddii" and a co-occurring Enterobacteriaceae bacterium for which we propose the name "Candidatus Psyllophila symbiotica." Its localization within the bacteriome and its small genome size confirm that Psyllophila is a co-primary endosymbiont widespread within the genus Cacopsylla. Despite its highly eroded genome, Psyllophila perfectly complements the tryptophan biosynthesis pathway that is incomplete in the co-occurring Carsonella. Moreover, the genome of Psyllophila is almost as small as Carsonella's, suggesting an ancient dual endosymbiosis that has now reached a precarious stage where any additional gene loss would make the system collapse. Hence, our results shed light on the dynamic interactions of psyllids and their endosymbionts over evolutionary time.

Seasonal wild dance of dual endosymbionts in the pear psyllid Cacopsylla pyricola (Hemiptera: Psylloidea).

Most sap-feeding insects maintain obligate relationships with endosymbiotic bacteria that provide their hosts with essential nutrients. However, knowledge about the dynamics of endosymbiont titers across seasons in natural host populations is scarce. Here, we used quantitative PCR to investigate the seasonal dynamics of the dual endosymbionts "Candidatus Carsonella ruddii" and "Ca. Psyllophila symbiotica" in a natural population of the pear psyllid Cacopsylla pyricola (Hemiptera: Psylloidea: Psyllidae). Psyllid individuals were collected across an entire year, covering both summer and overwintering generations. Immatures harboured the highest titers of both endosymbionts, while the lowest endosymbiont density was observed in males. The density of Carsonella remained high and relatively stable across the vegetative period of the pear trees, but significantly dropped during the non-vegetative period, overlapping with C. pyricola's reproductive diapause. In contrast, the titer of Psyllophila was consistently higher than Carsonella's and exhibited fluctuations throughout the sampling year, which might be related to host age. Despite a tightly integrated metabolic complementarity between Carsonella and Psyllophila, our findings highlight differences in their density dynamics throughout the year, that might be linked to their metabolic roles at different life stages of the host.

Wolbachia infection dynamics in a natural population of the pear psyllid Cacopsylla pyri (Hemiptera: Psylloidea) across its seasonal generations.

Wolbachia is one of the most abundant intracellular symbionts of arthropods and has profound effects on host biology. Wolbachia transmission and host phenotypes often depend on its density within the host, which can be affected by multiple biotic and abiotic factors. However, very few studies measured Wolbachia density in natural host populations. Here, we describe Wolbachia in the pear psyllid Cacopsylla pyri from three populations in the Czech Republic. Using phylogenetic analyses based on wsp and multilocus sequence typing genes, we demonstrate that C. pyri harbours three new Wolbachia strains from supergroup B. A fourth Wolbachia strain from supergroup A was also detected in parasitised immatures of C. pyri, but likely came from a hymenopteran parasitoid. To obtain insights into natural Wolbachia infection dynamics, we quantified Wolbachia in psyllid individuals from the locality with the highest prevalence across an entire year, spanning several seasonal generations of the host. All tested females were infected and Wolbachia density remained stable across the entire period, suggesting a highly efficient vertical transmission and little influence from the environment and different host generations. In contrast, we observed a tendency towards reduced Wolbachia density in males which may suggest sex-related differences in Wolbachia-psyllid interactions.

Microbiome of pear psyllids: A tale about closely related species sharing their endosymbionts.

Psyllids are phloem-feeding insects that can transmit plant pathogens such as phytoplasmas, intracellular bacteria causing numerous plant diseases worldwide. Their microbiomes are essential for insect physiology and may also influence the capacity of vectors to transmit pathogens. Using 16S rRNA gene metabarcoding, we compared the microbiomes of three sympatric psyllid species associated with pear trees in Central Europe. All three species are able to transmit 'Candidatus Phytoplasma pyri', albeit with different efficiencies. Our results revealed potential relationships between insect biology and microbiome composition that varied during psyllid ontogeny and between generations in Cacopsylla pyri and C. pyricola, as well as between localities in C. pyri. In contrast, no variations related to psyllid life cycle and geography were detected in C. pyrisuga. In addition to the primary endosymbiont Carsonella ruddii, we detected another highly abundant endosymbiont (unclassified Enterobacteriaceae). C. pyri and C. pyricola shared the same taxon of Enterobacteriaceae which is related to endosymbionts harboured by other psyllid species from various families. In contrast, C. pyrisuga carried a different Enterobacteriaceae taxon related to the genus Sodalis. Our study provides new insights into host-symbiont interactions in psyllids and highlights the importance of host biology and geography in shaping microbiome structure.

Systematics, biogeography and host-plant relationships of the Neotropical jumping plant-louse genus Russelliana (Hemiptera: Psylloidea).

The Neotropical genus Russelliana (Psyllidae: Aphalaroidinae) is revised and its phylogenetic, host-plant and biogeographical relationships are discussed. Twenty-four species are described as new, bringing the number of known species to 43. An identification key is provided for the adults. A phylogenetic analysis of 26 morphological characters resulted in 54 most parsimonious trees. The consensus tree is well resolved at the base but poorly at the crown. Most Russelliana species are monophagous or oligophagous with the exception of R. solanicola which is polyphagous. With eight plant families, the host range of Russelliana is broader than that of other aphalaroidine genera. The hosts for 29 species are confirmed, those for 12 species are suggested based on phylogenetic relationships. The species associated with Asteraceae (4 spp.) and most of the Fabaceae-feeders (12 spp.) form each a monophyletic group, those associated with Verbenaceae (5 spp.) are paraphyletic and those with Solanaceae are polyphyletic (16 spp.). The two species associated with Rosaceae are not closely related. These patterns suggest repeated host shifts. Whether there is cospeciation in some groups cannot be judged as neither psyllid nor host phylogenies are sufficiently resolved. The world psylloid fauna comprises relatively few species associated with Solanaceae. The number of 16 Russelliana species with confirmed or likely solanaceous hosts is, therefore, surprising and important in view of the potential pest status of some Russelliana spp. The genus is restricted to temperate and subtropical South America (Argentina, Bolivia, Southern Brazil, Chile, Peru and Uruguay). Most species are known from the Western Andean part of the continent. Only four species are currently known from Eastern South America. The cladogram suggests that geographical vicariance may account for at least part of the observed species richness, as five vicariant events were detected for Russelliana. A better resolution of the cladogram may reveal more cases of geographical vicariance.

Emerging New Crop Pests: Ecological Modelling and Analysis of the South American Potato Psyllid Russelliana solanicola (Hemiptera: Psylloidea) and Its Wild Relatives.

Food security is threatened by newly emerging pests with increased invasive potential accelerated through globalization. The Neotropical jumping plant louse Russelliana solanicola Tuthill is currently a localized potato pest and probable vector of plant pathogens. It is an unusually polyphagous species and is widely distributed in and along the Andes. To date, introductions have been detected in eastern Argentina, southern Brazil and Uruguay. Species distribution models (SDMs) and trait comparisons based on contemporary and historical collections are used to estimate the potential spread of R. solanicola worldwide. We also extend our analyses to all described species in the genus Russelliana in order to assess the value of looking beyond pest species to predict pest spread. We investigate the extent to which data on geographical range and environmental niche can be effectively extracted from museum collections for comparative analyses of pest and non-pest species in Russelliana. Our results indicate that R. solanicola has potential for invasion in many parts of the world with suitable environmental conditions that currently have or are anticipated to increase potato cultivation. Large geographical ranges are characteristic of a morphological subgeneric taxon group that includes R. solanicola; this same group also has a larger environmental breadth than other groups within the genus. Ecological modelling using museum collections provides a useful tool for identifying emerging pests and developing integrated pest management programs.

The potato pest Russelliana solanicola Tuthill (Hemiptera: Psylloidea): taxonomy and host-plant patterns.

The Neotropical jumping plant-louse Russelliana solanicola Tuthill is a potato pest and a probable vector of plant pathogens. Populations morphologically similar to those found on potatoes have been collected on plants of at least ten different families, four of which have been confirmed as hosts by the presence of immatures. This suggests that R. solanicola is either a single polyphagous species or a complex of closely related, monophagous species (host races/cryptic species). Results of our analyses of multiple morphometric characters show for both sexes a grouping of the populations of R. solanicola and a clear separation of the latter from other Russelliana species. On the other hand, within R. solanicola, there is an overlap of populations from different host-plants as well as from different geographical regions. The results of the present study strongly suggest that R. solanicola is a single, polyphagous species and the known distribution indicates that it is native to the Andes. It is likely that R. solanicola has been introduced into eastern Argentina, Brazil and Uruguay. The polyphagy together with the ability to disperse and transmit plant pathogens potentially make this species an economically important pest of potato and other crop species.