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Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR

Bibliographic Details
Main Author: Gouveia, Virgilio Pires
Publication Date: 2021
Format: Master thesis
Language: eng
Source: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Download full: http://hdl.handle.net/10400.5/23824
Summary: Mestrado em Engenharia Agronómica - Proteção de Plantas / Instituto Superior de Agronomia. Universidade de Lisboa
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spelling Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCRnematodePinus pinasterqPCRregressionquantificationMestrado em Engenharia Agronómica - Proteção de Plantas / Instituto Superior de Agronomia. Universidade de LisboaNative to Europe, the maritime pine (Pinus pinaster) spreads through mainland Portugal. Multiple economic agents depend on its existence, namely for construction wood, or resin. As the second largest forest species in occupied area, the pine´s importance does not restrict to the economy: Pinhal do Rei is of cultural relevance, and the enourmous biotic chain dependent on the pine forests is extremely important to protect, specially considering the portuguese climatic regime, unsuitable for many species. Besides forest fires, pests and diseases were also accountable for forest area reductions is the last years. Pine wilt disease (PWD) caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, has been particularly damaging. The exotic parasite uses a native vector for dissemination, and is unchallenged while spreading through healthy trees. Measures to reduce dissemination and control the spread of the associated PWD are of utter importance for all Europe, and they must de supported by reliable quantification and detection methods of PWN. Real-time quantitative PCR parameters were studied with samples from an experiment with three year-old inoculated pine seedlings, collected at five time-points. From 12 genes tested, two were considered adequate by conventional PCR. After PCR-product sequencing and validation, samples were tested in qPCR. A linear regression from knwon-concentration PWN samples created a calibration curve, while primer efficiency was obtained with a dilution series. BxITS2 was proved the most suitable gene, producing more reliable results when amplifying nematode and plant DNA altogether. The positive results of quantification on samples with parasite and plant DNA extracted simultaneously are very promising. Thus, we validated a direct extraction quantification method, but the methodology can still be improved to overcome identified weaknessesISA/ULMiguel, Célia RodriguesRamos, Ana PaulaRepositório da Universidade de LisboaGouveia, Virgilio Pires2025-03-01T01:30:38Z20212021-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.5/23824engGouveia, V.P. - Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR. Lisboa: ISA, 2021, 83 p.info:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2025-03-17T16:08:50Zoai:repositorio.ulisboa.pt:10400.5/23824Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-29T04:04:02.580423Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR
title Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR
spellingShingle Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR
Gouveia, Virgilio Pires
nematode
Pinus pinaster
qPCR
regression
quantification
title_short Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR
title_full Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR
title_fullStr Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR
title_full_unstemmed Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR
title_sort Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR
author Gouveia, Virgilio Pires
author_facet Gouveia, Virgilio Pires
author_role author
dc.contributor.none.fl_str_mv Miguel, Célia Rodrigues
Ramos, Ana Paula
Repositório da Universidade de Lisboa
dc.contributor.author.fl_str_mv Gouveia, Virgilio Pires
dc.subject.por.fl_str_mv nematode
Pinus pinaster
qPCR
regression
quantification
topic nematode
Pinus pinaster
qPCR
regression
quantification
description Mestrado em Engenharia Agronómica - Proteção de Plantas / Instituto Superior de Agronomia. Universidade de Lisboa
publishDate 2021
dc.date.none.fl_str_mv 2021
2021-01-01T00:00:00Z
2025-03-01T01:30:38Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10400.5/23824
url http://hdl.handle.net/10400.5/23824
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Gouveia, V.P. - Optimizing the quantification of the pine wood nematode (Bursaphelenchus xylophilus) in Pinus pinaster through real-time quantitative PCR. Lisboa: ISA, 2021, 83 p.
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv ISA/UL
publisher.none.fl_str_mv ISA/UL
dc.source.none.fl_str_mv reponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron:RCAAP
instname_str FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron_str RCAAP
institution RCAAP
reponame_str Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
collection Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
repository.name.fl_str_mv Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
repository.mail.fl_str_mv info@rcaap.pt
_version_ 1833601930983636992

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