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A multi-tool approach to assess microalgal diversity in lichens. isolation, Sanger sequencing, HTS and ultrastructural correlations

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A multi-tool approach to assess microalgal diversity in lichens. isolation, Sanger sequencing, HTS and ultrastructural correlations

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dc.contributor.author Molins, Arantxa es_ES
dc.contributor.author Moya, P. es_ES
dc.contributor.author García-Breijo, Francisco-José es_ES
dc.contributor.author Reig-Armiñana, José es_ES
dc.contributor.author Barreno, Eva es_ES
dc.date.accessioned 2019-06-08T20:03:00Z
dc.date.available 2019-06-08T20:03:00Z
dc.date.issued 2018 es_ES
dc.identifier.issn 0024-2829 es_ES
dc.identifier.uri http://hdl.handle.net/10251/121824
dc.description.abstract [EN] Lichen thalli represent the most conspicuous examples of fungal-algal interactions. Studies that describe phycobiont diversity within entire thalli are based mainly on Sanger sequencing. In some lichen species, this technique could underestimate the intrathalline coexistence of multiple microalgae. In this study different multi-tool approaches were applied to two lichen taxa, Circinaria hispida and Flavoparmelia soredians, to detect algal coexistence. Here, we combined Sanger sequencing, a specific polymerase chain reaction (PCR) primer, 454-pyrosequencing, phycobiont isolation and ultrastructural characterization. Furthermore, we compared pyrenoid ultrastructural features of lichenized phycobionts with microalgae isolated in culture. An improved methodology was used to isolate and propagate phycobionts which, in combination with fast genetic identification, resulted in a considerable reduction in time and cost to complete the process. This isolation method, coupled with a specific PCR primer, allowed for the detection of coexisting algae in C. hispida (four Trebouxia lineages). 454-pyrosequencing detected only a fraction of such diversity, while Sanger sequencing identified only the primary phycobiont. Ultrastructural features of the isolated algae were observed by transmission electron microscopy; the maintenance of the pyrenoid characteristics suggested the existence of different Trebouxia lineages. In F. soredians a single Trebouxia lineage was identified using all these approaches. In cases of lichens with algal coexistence, a combination of different molecular and ultrastructural approaches may be required to reveal the underlying algal diversity within a single thallus. The approach proposed in this study provides information about the relationship between molecular and ultrastructural data, and represents an improvement in the delimitation of taxonomic features which is needed to recognize intrathalline Trebouxia diversity. es_ES
dc.description.sponsorship Supported by the Ministerio de Economia y Competitividad (MINECO, Spain) (CGL2016-79158-P) and Excellence in Research (Generalitat Valenciana, Spain) (PROMETEOII/2013/021 and PROMETEO/2017/039). We wish to thank the technicians (Maria Teresa Minguez and Nuria Cebrian) of the Servicio de Microscopia Electronica, SCSIE and Jardi Botanic (Universitat de Valencia) who helped us to perform the TEM process, and Santiago Catala for the pyrosequencing analyses. Daniel Sheerin revised the English manuscript. es_ES
dc.language Inglés es_ES
dc.publisher Cambridge University Press es_ES
dc.relation GV/PROMETEOII/2013/021 es_ES
dc.relation MICINN/FEDER CGL2016-79158-P es_ES
dc.relation GV/PROMETEO/2017/039 es_ES
dc.relation.ispartof The Lichenologist es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject Coexistence es_ES
dc.subject Propagation es_ES
dc.subject 454-pyrosequencing es_ES
dc.subject Trebouxia es_ES
dc.subject.classification BOTANICA es_ES
dc.title A multi-tool approach to assess microalgal diversity in lichens. isolation, Sanger sequencing, HTS and ultrastructural correlations es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1017/S0024282917000664 es_ES
dc.rights.accessRights Abierto es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Ecosistemas Agroforestales - Departament d'Ecosistemes Agroforestals es_ES
dc.description.bibliographicCitation Molins, A.; Moya, P.; García-Breijo, F.; Reig-Armiñana, J.; Barreno, E. (2018). A multi-tool approach to assess microalgal diversity in lichens. isolation, Sanger sequencing, HTS and ultrastructural correlations. The Lichenologist. 50(1):123-138. https://doi.org/10.1017/S0024282917000664 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion http://doi.org/10.1017/S0024282917000664 es_ES
dc.description.upvformatpinicio 123 es_ES
dc.description.upvformatpfin 138 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 50 es_ES
dc.description.issue 1 es_ES
dc.relation.pasarela 339371 es_ES
dc.contributor.funder Ministerio de Ciencia, Innovación y Universidades es_ES
dc.contributor.funder Generalitat Valenciana es_ES


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