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Pollen identification by its2 metabarcoding: curation of the sequences retrieved from genbank to build a reference database

dc.contributor.authorQuaresma, Andreia
dc.contributor.authorKeller, Alexander
dc.contributor.authorRufino, José
dc.contributor.authorSteen, Jozef van der
dc.contributor.authorPinto, M. Alice
dc.date.accessioned2023-05-22T11:14:11Z
dc.date.available2023-05-22T11:14:11Z
dc.date.issued2022
dc.description.abstractA powerful way of studying the quality of the environment is by examining the pollen collected by honey bees (Apis mellifera) as it contains information on available plant sources, spatial and temporal floral diversity, as well as on chemical contaminants. This entails botanical identification of pollen which has typically been addressed by classical palynology, a costly approach that often provides low taxonomic resolution, is time-consuming, labour intensive, and requires plant taxonomy expertise. However, with high-throughput sequencing becoming increasingly affordable, pollen metabarcoding is gaining momentum, and it is a promising alternative to classical palynology. But one of the main drawbacks of pollen metabarcoding is the lack of good quality reference databases for the barcode of choice. BCdatabaser (Keller et al. 2020) was developed to automatically generate a standardized database for the ITS2 barcode from the primary sequence database GenBank. While using BCdatabaser to construct an ITS2 reference database for identification of bee-collected pollen, we noticed several misidentified sequences retrieved from GenBank, which would impact identification accuracy. There were two types of problems: plant sequences that were assigned to the wrong plant species and fungi sequences that were identified as plants. To overcome these issues, we developed scripts in bash and R to curate an ITS2 reference database for pollen identification purposes. These scripts allowed us to identify the Fungi sequences retrieved from GenBank for subsequent removal from the database, to perform a pairwise alignment of all the sequences using vsearch v2.14.1 (Rognes et al., 2016) and, then to remove all the sequences with low identity percentage using an iteration process in R v4.1.2. The database curation is automated therefore enabling easy update of the ITS2 database to take advantage of the new sequences that are regularly deposited in GenBank.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationQuaresma, Andreia; Keller, Alexander; Rufino, José; Van der Steen, Jozef; Pinto, M. Alice (2022). Pollen identification by its2 metabarcoding: curation of the sequences retrieved from genbank to build a reference database. In EurBee 9 - 9th European Congress of Apidology. Belgrade, Serbiapt_PT
dc.identifier.isbn978-86-7078-173-3
dc.identifier.urihttp://hdl.handle.net/10198/28341
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherUniversity of Belgrade, Faculty of Biologypt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectPollen DNA metabarcodingpt_PT
dc.subjectITS2 database curationpt_PT
dc.subjectHoneybeept_PT
dc.titlePollen identification by its2 metabarcoding: curation of the sequences retrieved from genbank to build a reference databasept_PT
dc.typeconference object
dspace.entity.typePublication
oaire.citation.conferencePlaceBelgrade, Serbiapt_PT
oaire.citation.titleEurBee 9 - 9th European Congress of Apidologypt_PT
person.familyNameQuaresma
person.familyNameRufino
person.familyNamePinto
person.givenNameAndreia
person.givenNameJosé
person.givenNameM. Alice
person.identifier.ciencia-id4F1A-4E4A-3F23
person.identifier.ciencia-idC414-F47F-6323
person.identifier.ciencia-idF814-A1D0-8318
person.identifier.orcid0000-0002-8678-5800
person.identifier.orcid0000-0002-1344-8264
person.identifier.orcid0000-0001-9663-8399
person.identifier.scopus-author-id57119742600
person.identifier.scopus-author-id55947199100
person.identifier.scopus-author-id8085507800
rcaap.rightsopenAccesspt_PT
rcaap.typeconferenceObjectpt_PT
relation.isAuthorOfPublicationd417b0ac-c8ee-473a-a355-820b5b9a3f55
relation.isAuthorOfPublication1e24d2ce-a354-442a-bef8-eebadd94b385
relation.isAuthorOfPublication0667fe04-7078-483d-9198-56d167b19bc5
relation.isAuthorOfPublication.latestForDiscovery1e24d2ce-a354-442a-bef8-eebadd94b385

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