Abstract
Symbioses between plants and nitrogen-fixing cyanobacteria are benchmark biological systems to understand mutualism and patterns of coevolution. The diversity of cyanobacteria associated to non-vascular land plants is still being unraveled, with a huge gap in knowledge from tropical areas, especially in hornworts. We focus on the Neotropical hornwort Leiosporoceros dussii, which has a semi-permanent association with cyanobacteria. Unlike other hornworts, the peculiar Nostoc canals suggest cyanobiont specificity and selectivity. Here, we characterize the first hornwort bacteriome and focus on the endophytic cyanobacteria by describing its morphology, metagenomic diversity and phylogeny between two sampling sites in Panama. First, we use a metagenomic analyses (16S) from gametophytes from Río El Guayabo and Río Indio in Panama to identify the endophytic bacteria community of L. dussii. To provide more information on the cyanobacterial endophytes, we have extracted cyanobacterial endophytes to measure cell size variation and use the markers trnL and rbcLX to determine their phylogenetic relationships. Finally, we use stable isotope δ 15N to assess nitrogen flow in the plants. We find little diversity in Leiosporoceros bacteriome but observe significant community variations between sites. Results indicate a lack of cyanobacterial specificity of cyanobacteria associated to Leiosporoceros, with six unrelated clades of Nostoc forming a polyphyletic assemblage. Four clades are nested within bacteria collected as free-living taxa suggesting that typically non-symbiotic cyanobacteria can be associated with this hornwort. Finally, the little nitrogen fractioning of the plant suggests the role of the cyanobacterial symbionts in nitrogen fixation. This study provides the first hornwort bacteriome and is an important step in the characterization of tropical bryophyte symbioses.
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Data availability
The dataset generated and analysed during the current study is available from GenBank and DRYAD (https://doi.org/10.5061/dryad.sxksn030k).
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Funding
Établissement de nouveaux chercheurs universitaires FRQNT- 206943 to JCVA.
Earl S. Tupper Fellowship 2015 to JCVA.
National Science Foundation Dimensions of Biodiversity grant (1831428) to F.-W.L.
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RB and JCV conceived the study and performed the phylogenetic work and wrote the first version of the manuscript; GPB did the metagenomic analysis; ST performed the analysis of isotope data, YG did the morphological measurements, JG and NSA provided logistic support in the field and funding; FWL contributed with logistic support and funding; RB, GPB, NS, FWL, ST and JCV wrote and edited the manuscript.
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Bouchard, R., Peñaloza-Bojacá, G., Toupin, S. et al. Contrasting bacteriome of the hornwort Leiosporoceros dussii in two nearby sites with emphasis on the hornwort-cyanobacterial symbiosis. Symbiosis 81, 39–52 (2020). https://doi.org/10.1007/s13199-020-00680-1
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DOI: https://doi.org/10.1007/s13199-020-00680-1