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Bacterial endosymbionts of Placobdella (Annelida: Hirudinea: Glossiphoniidae): phylogeny, genetic distance, and vertical transmission

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Abstract

Blood-feeding leeches of the genus Placobdella have acquired intracellular alphaproteobacterial endosymbionts of the genus Reichenowia that potentially aid in the production of B vitamins, thereby ameliorating the lack of these essential nutrients in the diet of the hosts. For Placobdella associates, little is still known about the diversity, genetic makeup, and the mode of transmission of bacteria between leeches. We aimed to (i) place the bacterial symbionts in a phylogenetic context, (ii) compare patterns of cladogenesis between the bacteria and hosts to search for evidence of co-speciation, and (iii) explore the mode of bacterial transmission between leech parent and offspring. DNA sequencing of the bacterial 16S rDNA and 23S rDNA loci suggests that, whereas Reichenowia forms a monophyletic group within the alphaproteobacterial family Rhizobiaceae, no evidence for co-speciation between hosts and bacteria can be traced. Attempts at DNA amplification for ovarial tissues were negative for a range of species, but two 16S rDNA sequences retrieved from the testisacs of P. rugosa showed very high similarity with Reichenowia. Although we cannot rule out that this may be a contamination, or a different, potentially free-living species of bacteria, our results may indicate that Reichenowia is transferred from leech parent to offspring via the testisacs.

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Acknowledgements

We thank Steven Doyle for aiding with imaging, Michael Tessler for creating the script used to run TNT, and the Willi Hennig Society for making TNT freely available. Danielle de Carle and Rafael Iwama provided much input during analyses and interpretation of the results, which greatly benefitted the paper. We thank the Saskatchewan ministry of Parks, Culture, and Sports, the Ontario Ministry of the Environment, Conservation, and Parks, as well as Manitoba Parks for help with permitting. This research was funded by a NSERC Discovery grant and an Olle Engkvist Byggmästare stipend to SK.

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  1. Department of Natural History, Royal Ontario Museum, 100 Queen’s Park, Toronto, ON, M5S 2C6, Canada

    Claire Manglicmot & Sebastian Kvist

  2. Department of Ecology and Evolutionary Biology, University of Toronto, 25 Willcocks Street, Toronto, ON, M5S 2B4, Canada

    Claire Manglicmot & Sebastian Kvist

  3. Laboratorio de Helmintología, Instituto de Biología, Universidad Nacional Autónoma de México, Ciudad Universitaria, Copilco, Coyoacán, A.P. 70-153, C.P. 14510, Mexico City, Mexico

    Alejandro Oceguera-Figueroa

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10750_2019_4175_MOESM1_ESM.tif

Supplementary material 1 (TIFF 4222 kb) Supplementary Fig. S1. Phylogenetic tree based on 16S and 23S rDNA resulting from the parsimony analysis (17,397 steps, consistency index = 0.344, retention index = 0.675). Bootstrap support values are shown above each node, and branch lengths are drawn proportional to the amount of change

10750_2019_4175_MOESM2_ESM.tif

Supplementary material 2 (TIFF 2971 kb) Supplementary Fig. S2. Phylogenetic tree based on 16S and 23S rDNA resulting from the Bayesian inference analysis. Posterior probabilities are shown as above each node, and branch lengths are drawn proportional to the amount of change

10750_2019_4175_MOESM3_ESM.tif

Supplementary material 3 (TIFF 1231 kb) Supplementary Fig. S3. Co-phylogeny solution map based on approximately 1,000 solutions for each of the event-cost schemes. The figure shows one of several isomorphic tanglemaps generated for the “01210 + time zones” scheme. Note that the earliest bifurcation of the host and symbiont trees begin at the same “timezone” such that their earlier nodes on the tree ‘overlap’. The ancestor to ex. P. hollensis, R. parasiticae, ex. P. lamothei, and ex. P. papillifera tend to be shown as horizontal transmission donors of duplicate lineages in other event-cost schemes as well (data not shown). These lineages also tend to undergo duplication and host-switching in other event-cost schemes

10750_2019_4175_MOESM4_ESM.tif

Supplementary material 4 (TIFF 3133 kb) Supplementary Fig. S4. Host-constrained maximum likelihood phylogeny resulting from the analysis of the concatenated 16S and 23S matrix. The symbiont topology was constrained to the host Placobdella phylogeny published by de Carle et al. (2016). Bootstrap support values > 50% are shown above each node. Branch lengths are drawn proportional to the amount of change. The final log likelihood score for this tree was -81439.0

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Manglicmot, C., Oceguera-Figueroa, A. & Kvist, S. Bacterial endosymbionts of Placobdella (Annelida: Hirudinea: Glossiphoniidae): phylogeny, genetic distance, and vertical transmission. Hydrobiologia 847, 1177–1194 (2020). https://doi.org/10.1007/s10750-019-04175-z

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