Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , , , |
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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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|| |
_version_ |
1797069079807262720 |