Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA

Detalhes bibliográficos
Autor(a) principal: Azêvedo, Hellen Sandra Freires da Silva
Data de Publicação: 2019
Outros Autores: Rufino, Polinar Bandeira, Azevedo, José Marlo Araújo de, Silva, Luciélio Manoel da, Wadt, Lúcia Helena de Oliveira, Campos, Tatiana de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Bioscience journal (Online)
Texto Completo: https://seer.ufu.br/index.php/biosciencejournal/article/view/42190
Resumo: The objective of this study was to test the efficiency of preservation and maceration methods for Euterpe precatoria leaflet tissue to obtain genomic DNA for molecular studies. The leaflets of E. precatoria were collected in an experimental field at Embrapa Acre, Brazil. The study was conducted in a completely randomized design with 10 replicates, in a 12 × 2 factorial structure, with 12 storage treatments (fresh; lyophiliser 3 days; refrigerator 3, 5, and 7 days; silica gel 7, 10, 20, and 30 days; and transport buffer 3, 5, and 7 days) and two leaf tissue maceration methods (liquid nitrogen and the TissueLyser®). Statistically significant differences in the obtained DNA concentration were found between the maceration and storage treatments. The TissueLyser® macerator produced higher DNA concentrations when compared to liquid nitrogen. For the storage treatments, five groups were formed based on DNA concentration when macerated with the TissueLyser® and two groups when macerated with liquid nitrogen. The DNA concentrations ranged from 285.00 ng/µL (7 days in transport buffer) to 702.00 ng/µL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/µL (30 days in silica gel) to 2,850.00 ng/µL (3 days in lyophiliser) using the TissueLyser® macerator. The DNA purity (A260/A280 nm) varied from 1.30 to 1.70 when the leaflets were macerated with liquid nitrogen and from 1.30 to 1.90 with the TissueLyser® macerator. Despite the variations in leaf tissue preservation and DNA concentration, all treatments were effective for DNA isolation and it was possible to amplify genomic regions of microsatellite markers by PCR. It was concluded that leaflets of E. precatoria stored in a lyophiliser and processed with an automatic macerator resulted in satisfactory DNA for molecular studies.
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spelling Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNAPreservação e maceração de tecido de folíolos de açaí da Amazônia para obtenção de DNA genômicoAçaí-solteiroCTAB methodDNA isolationEuterpe precatoriaBiological SciencesThe objective of this study was to test the efficiency of preservation and maceration methods for Euterpe precatoria leaflet tissue to obtain genomic DNA for molecular studies. The leaflets of E. precatoria were collected in an experimental field at Embrapa Acre, Brazil. The study was conducted in a completely randomized design with 10 replicates, in a 12 × 2 factorial structure, with 12 storage treatments (fresh; lyophiliser 3 days; refrigerator 3, 5, and 7 days; silica gel 7, 10, 20, and 30 days; and transport buffer 3, 5, and 7 days) and two leaf tissue maceration methods (liquid nitrogen and the TissueLyser®). Statistically significant differences in the obtained DNA concentration were found between the maceration and storage treatments. The TissueLyser® macerator produced higher DNA concentrations when compared to liquid nitrogen. For the storage treatments, five groups were formed based on DNA concentration when macerated with the TissueLyser® and two groups when macerated with liquid nitrogen. The DNA concentrations ranged from 285.00 ng/µL (7 days in transport buffer) to 702.00 ng/µL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/µL (30 days in silica gel) to 2,850.00 ng/µL (3 days in lyophiliser) using the TissueLyser® macerator. The DNA purity (A260/A280 nm) varied from 1.30 to 1.70 when the leaflets were macerated with liquid nitrogen and from 1.30 to 1.90 with the TissueLyser® macerator. Despite the variations in leaf tissue preservation and DNA concentration, all treatments were effective for DNA isolation and it was possible to amplify genomic regions of microsatellite markers by PCR. It was concluded that leaflets of E. precatoria stored in a lyophiliser and processed with an automatic macerator resulted in satisfactory DNA for molecular studies.O objetivo deste estudo foi testar a eficiência de métodos de preservação e maceração de tecidos de folíolos de Euterpe precatoria para obter DNA genômico para estudos moleculares. Os folíolos de E. precatoria foram coletados no campo experimental da Embrapa Acre, Brasil. O estudo foi conduzido em delineamento inteiramente casualizado com 10 repetições, em esquema fatorial 12 × 2, com 12 tratamentos de armazenamento (fresco; liofilizado 3 dias; geladeira 3, 5, e 7; sílica gel 7, 10, 20 e 30 dias e tampão de transporte 3, 5 e 7 dias) e dois tipos de maceração do tecido foliar (nitrogênio líquido e TissueLyser®). Para a variável concentração de DNA houve diferença estatística entre os tipos maceração e de armazenamento. O macerador TissueLyser® apresentou maiores concentrações de DNA quando comparado ao nitrogênio líquido. Para os tipos de armazenamento verificou-se formação de cinco grupos quando macerados TissueLyser® e dois grupos quando macerados com nitrogênio líquido. As concentrações de DNA variaram de 285,00 ng/µL (7 dias em tampão de transporte) a 702,00 ng/µL (30 dias em sílica gel) quando maceradas com nitrogênio líquido. Quando maceradas com macerador TissueLyser® variaram de 572,73 ng/µL (30 dias em sílica gel) a 2.850,00 ng/µL (3 dias em liofilizador). A pureza do DNA (A260/A280 nm) variou de 1,30 a 1,70 quando os folíolos foram macerados com nitrogênio líquido e de 1,30 a 1,90 quando macerados em macerador TissueLyser®. Apesar das variações na conservação e concentração dos tecidos foliares, todos os tratamentos foram eficazes para o isolamento do DNA e amplificaram regiões de marcadores microssatélites. Concluiu-se que folíolos de E. precatoria armazenados em liofilizador e macerados com macerador automático resultaram em DNA satisfatório para estudos moleculares.EDUFU2019-08-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://seer.ufu.br/index.php/biosciencejournal/article/view/4219010.14393/BJ-v35n4a2019-42190Bioscience Journal ; Vol. 35 No. 4 (2019): July/Aug.; 1188-1197Bioscience Journal ; v. 35 n. 4 (2019): July/Aug.; 1188-11971981-3163reponame:Bioscience journal (Online)instname:Universidade Federal de Uberlândia (UFU)instacron:UFUenghttps://seer.ufu.br/index.php/biosciencejournal/article/view/42190/26657Brazil; ContemporaryCopyright (c) 2019 Hellen Sandra Freires da Silva Azêvedo, Polinar Bandeira Rufino, José Marlo Araújo de Azevedo, Luciélio Manoel da Silva, Lúcia Helena de Oliveira Wadt, Tatiana de Camposhttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessAzêvedo, Hellen Sandra Freires da SilvaRufino, Polinar BandeiraAzevedo, José Marlo Araújo deSilva, Luciélio Manoel daWadt, Lúcia Helena de OliveiraCampos, Tatiana de2022-01-31T00:29:03Zoai:ojs.www.seer.ufu.br:article/42190Revistahttps://seer.ufu.br/index.php/biosciencejournalPUBhttps://seer.ufu.br/index.php/biosciencejournal/oaibiosciencej@ufu.br||1981-31631516-3725opendoar:2022-01-31T00:29:03Bioscience journal (Online) - Universidade Federal de Uberlândia (UFU)false
dc.title.none.fl_str_mv Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA
Preservação e maceração de tecido de folíolos de açaí da Amazônia para obtenção de DNA genômico
title Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA
spellingShingle Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA
Azêvedo, Hellen Sandra Freires da Silva
Açaí-solteiro
CTAB method
DNA isolation
Euterpe precatoria
Biological Sciences
title_short Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA
title_full Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA
title_fullStr Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA
title_full_unstemmed Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA
title_sort Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA
author Azêvedo, Hellen Sandra Freires da Silva
author_facet Azêvedo, Hellen Sandra Freires da Silva
Rufino, Polinar Bandeira
Azevedo, José Marlo Araújo de
Silva, Luciélio Manoel da
Wadt, Lúcia Helena de Oliveira
Campos, Tatiana de
author_role author
author2 Rufino, Polinar Bandeira
Azevedo, José Marlo Araújo de
Silva, Luciélio Manoel da
Wadt, Lúcia Helena de Oliveira
Campos, Tatiana de
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Azêvedo, Hellen Sandra Freires da Silva
Rufino, Polinar Bandeira
Azevedo, José Marlo Araújo de
Silva, Luciélio Manoel da
Wadt, Lúcia Helena de Oliveira
Campos, Tatiana de
dc.subject.por.fl_str_mv Açaí-solteiro
CTAB method
DNA isolation
Euterpe precatoria
Biological Sciences
topic Açaí-solteiro
CTAB method
DNA isolation
Euterpe precatoria
Biological Sciences
description The objective of this study was to test the efficiency of preservation and maceration methods for Euterpe precatoria leaflet tissue to obtain genomic DNA for molecular studies. The leaflets of E. precatoria were collected in an experimental field at Embrapa Acre, Brazil. The study was conducted in a completely randomized design with 10 replicates, in a 12 × 2 factorial structure, with 12 storage treatments (fresh; lyophiliser 3 days; refrigerator 3, 5, and 7 days; silica gel 7, 10, 20, and 30 days; and transport buffer 3, 5, and 7 days) and two leaf tissue maceration methods (liquid nitrogen and the TissueLyser®). Statistically significant differences in the obtained DNA concentration were found between the maceration and storage treatments. The TissueLyser® macerator produced higher DNA concentrations when compared to liquid nitrogen. For the storage treatments, five groups were formed based on DNA concentration when macerated with the TissueLyser® and two groups when macerated with liquid nitrogen. The DNA concentrations ranged from 285.00 ng/µL (7 days in transport buffer) to 702.00 ng/µL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/µL (30 days in silica gel) to 2,850.00 ng/µL (3 days in lyophiliser) using the TissueLyser® macerator. The DNA purity (A260/A280 nm) varied from 1.30 to 1.70 when the leaflets were macerated with liquid nitrogen and from 1.30 to 1.90 with the TissueLyser® macerator. Despite the variations in leaf tissue preservation and DNA concentration, all treatments were effective for DNA isolation and it was possible to amplify genomic regions of microsatellite markers by PCR. It was concluded that leaflets of E. precatoria stored in a lyophiliser and processed with an automatic macerator resulted in satisfactory DNA for molecular studies.
publishDate 2019
dc.date.none.fl_str_mv 2019-08-08
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://seer.ufu.br/index.php/biosciencejournal/article/view/42190
10.14393/BJ-v35n4a2019-42190
url https://seer.ufu.br/index.php/biosciencejournal/article/view/42190
identifier_str_mv 10.14393/BJ-v35n4a2019-42190
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://seer.ufu.br/index.php/biosciencejournal/article/view/42190/26657
dc.rights.driver.fl_str_mv https://creativecommons.org/licenses/by/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/4.0
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.coverage.none.fl_str_mv Brazil; Contemporary
dc.publisher.none.fl_str_mv EDUFU
publisher.none.fl_str_mv EDUFU
dc.source.none.fl_str_mv Bioscience Journal ; Vol. 35 No. 4 (2019): July/Aug.; 1188-1197
Bioscience Journal ; v. 35 n. 4 (2019): July/Aug.; 1188-1197
1981-3163
reponame:Bioscience journal (Online)
instname:Universidade Federal de Uberlândia (UFU)
instacron:UFU
instname_str Universidade Federal de Uberlândia (UFU)
instacron_str UFU
institution UFU
reponame_str Bioscience journal (Online)
collection Bioscience journal (Online)
repository.name.fl_str_mv Bioscience journal (Online) - Universidade Federal de Uberlândia (UFU)
repository.mail.fl_str_mv biosciencej@ufu.br||
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