Molecular diagnosis of trichodorid species from Portugal
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/1822/15350 |
Resumo: | A PCR-RFLP assay was developed for the identification of trichodorids belonging to Nanidorus, Paratrichodorus and Trichodorus genera. Exploring the variability of the 18S SSU rDNA gene this method provides for the first time a molecular diagnosis tool alternative to the difficult and time consuming phenotyping especially of quite similar species, enabling selectivity in mixed samples with non-trichodorid species, and also the differentiation of juveniles. Based on the alignment of previously obtained 18S rDNA nucleotide sequences of trichodorids from Portugal, a pair of selective primers was designed in conserved regions to allow the amplification in all known Portuguese species of a variable region located at the 3’ end of the gene. The PCR product, 615bp in length, exhibits nucleotide variability to generate restriction fragment patterns which were consistent among populations of the same species and allowed to discriminate trichodorids at the species level. The proposed protocol was tested and proved effective with twelve trichodorid species from Portugal (N. minor, P. allius, P. anemones, P. divergens, P. hispanus, P. pachydermus, P. porosus, T. beirensis, T. lusitanicus, T. primitivus and two other Trichodorus species, A and B) and six non-indigenous trichodorid populations (N. minor, P. allius, P. anemones, P. pachydermus, P. porosus, T. primitivus). |
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Molecular diagnosis of trichodorid species from PortugalIdentificationIntraspecific variationPCR-RFLPRibosomal DNATrichodorus spp.18S sequenceParatrichodorus spp.Nanidorus minor18S sequence identificationScience & TechnologyA PCR-RFLP assay was developed for the identification of trichodorids belonging to Nanidorus, Paratrichodorus and Trichodorus genera. Exploring the variability of the 18S SSU rDNA gene this method provides for the first time a molecular diagnosis tool alternative to the difficult and time consuming phenotyping especially of quite similar species, enabling selectivity in mixed samples with non-trichodorid species, and also the differentiation of juveniles. Based on the alignment of previously obtained 18S rDNA nucleotide sequences of trichodorids from Portugal, a pair of selective primers was designed in conserved regions to allow the amplification in all known Portuguese species of a variable region located at the 3’ end of the gene. The PCR product, 615bp in length, exhibits nucleotide variability to generate restriction fragment patterns which were consistent among populations of the same species and allowed to discriminate trichodorids at the species level. The proposed protocol was tested and proved effective with twelve trichodorid species from Portugal (N. minor, P. allius, P. anemones, P. divergens, P. hispanus, P. pachydermus, P. porosus, T. beirensis, T. lusitanicus, T. primitivus and two other Trichodorus species, A and B) and six non-indigenous trichodorid populations (N. minor, P. allius, P. anemones, P. pachydermus, P. porosus, T. primitivus).Centro de Biolgogia Molecular e Ambiental (CBMA)Fundação para a Ciência e a Tecnologia (FCT)Wiley-BlackwellUniversidade do MinhoDuarte, I. M.Almeida, M. T. M.Duarte, Maria MargaridaBrown, D. J. F.Neilson, R2011-062011-06-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/15350eng1365-305910.1111/j.1365-3059.2010.02401.xwileyonlinelibrary.cominfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:27:28Zoai:repositorium.sdum.uminho.pt:1822/15350Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:22:03.701285Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Molecular diagnosis of trichodorid species from Portugal |
title |
Molecular diagnosis of trichodorid species from Portugal |
spellingShingle |
Molecular diagnosis of trichodorid species from Portugal Duarte, I. M. Identification Intraspecific variation PCR-RFLP Ribosomal DNA Trichodorus spp. 18S sequence Paratrichodorus spp. Nanidorus minor 18S sequence identification Science & Technology |
title_short |
Molecular diagnosis of trichodorid species from Portugal |
title_full |
Molecular diagnosis of trichodorid species from Portugal |
title_fullStr |
Molecular diagnosis of trichodorid species from Portugal |
title_full_unstemmed |
Molecular diagnosis of trichodorid species from Portugal |
title_sort |
Molecular diagnosis of trichodorid species from Portugal |
author |
Duarte, I. M. |
author_facet |
Duarte, I. M. Almeida, M. T. M. Duarte, Maria Margarida Brown, D. J. F. Neilson, R |
author_role |
author |
author2 |
Almeida, M. T. M. Duarte, Maria Margarida Brown, D. J. F. Neilson, R |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Duarte, I. M. Almeida, M. T. M. Duarte, Maria Margarida Brown, D. J. F. Neilson, R |
dc.subject.por.fl_str_mv |
Identification Intraspecific variation PCR-RFLP Ribosomal DNA Trichodorus spp. 18S sequence Paratrichodorus spp. Nanidorus minor 18S sequence identification Science & Technology |
topic |
Identification Intraspecific variation PCR-RFLP Ribosomal DNA Trichodorus spp. 18S sequence Paratrichodorus spp. Nanidorus minor 18S sequence identification Science & Technology |
description |
A PCR-RFLP assay was developed for the identification of trichodorids belonging to Nanidorus, Paratrichodorus and Trichodorus genera. Exploring the variability of the 18S SSU rDNA gene this method provides for the first time a molecular diagnosis tool alternative to the difficult and time consuming phenotyping especially of quite similar species, enabling selectivity in mixed samples with non-trichodorid species, and also the differentiation of juveniles. Based on the alignment of previously obtained 18S rDNA nucleotide sequences of trichodorids from Portugal, a pair of selective primers was designed in conserved regions to allow the amplification in all known Portuguese species of a variable region located at the 3’ end of the gene. The PCR product, 615bp in length, exhibits nucleotide variability to generate restriction fragment patterns which were consistent among populations of the same species and allowed to discriminate trichodorids at the species level. The proposed protocol was tested and proved effective with twelve trichodorid species from Portugal (N. minor, P. allius, P. anemones, P. divergens, P. hispanus, P. pachydermus, P. porosus, T. beirensis, T. lusitanicus, T. primitivus and two other Trichodorus species, A and B) and six non-indigenous trichodorid populations (N. minor, P. allius, P. anemones, P. pachydermus, P. porosus, T. primitivus). |
publishDate |
2011 |
dc.date.none.fl_str_mv |
2011-06 2011-06-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/1822/15350 |
url |
http://hdl.handle.net/1822/15350 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1365-3059 10.1111/j.1365-3059.2010.02401.x wileyonlinelibrary.com |
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 |
Wiley-Blackwell |
publisher.none.fl_str_mv |
Wiley-Blackwell |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
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1799132689960271872 |