Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat
Autor(a) principal: | |
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Data de Publicação: | 2022 |
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/54513 |
Resumo: | Drought is one of the major constraints in durum wheat production in the Mediterranean Basin. In order to overcome this problem, the genetic transformation of durum wheat is one of the choices for improvement. However, the recalcitrance to Agrobacterium-mediated transformation in durum wheat (Triticum turgidum L.) is one of the factors limiting a successful genetic transformation. The aim of this study was to investigate the effect of explant type and acetosyringone concentration for the efficient Agrobacterium-mediated genetic transformation of three Moroccan durum wheat varieties (Amria, Chaoui, and Marouane). The mature embryos (intact, halved and pieces) were inoculated with Agrobacterium tumefaciens strain EHA101 harboring the binary vector pTF101.1 containing drought tolerance gene HVA1 from barley, and a selectable marker phosphinothricin (PPT) resistance (bar) gene. The explants were inoculated with A. tumefaciens (cell density OD650 at 0.7) at four different concentrations of acetosyringone (0, 100, 200, and 400 µM). The results showed that embryogenic calli from mature embryos showed higher regeneration and transformation than mature embryo halves and pieces. The integration of the transgene was confirmed by PCR amplification using primers specific to the bar gene, 2x35S promoter, and HVA1 gene. The transformation efficiency ranging from 0.33% to 2.33% was obtained in Amira variety using embryogenic calli and acetosyringone concentrations of 200 and 400 µM. The integration, as well as inheritance of the transgene, was confirmed by PCR amplification in T0 and T1 generations. This is the first report describing a genetic transformation of Moroccan durum wheat varieties via Agrobacterium tumefaciens. |
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Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheatAgrobacterium tumefaciensDrought ToleranceGenetic TransformationHVA1 GeneTriticum turgidum L.Agricultural SciencesDrought is one of the major constraints in durum wheat production in the Mediterranean Basin. In order to overcome this problem, the genetic transformation of durum wheat is one of the choices for improvement. However, the recalcitrance to Agrobacterium-mediated transformation in durum wheat (Triticum turgidum L.) is one of the factors limiting a successful genetic transformation. The aim of this study was to investigate the effect of explant type and acetosyringone concentration for the efficient Agrobacterium-mediated genetic transformation of three Moroccan durum wheat varieties (Amria, Chaoui, and Marouane). The mature embryos (intact, halved and pieces) were inoculated with Agrobacterium tumefaciens strain EHA101 harboring the binary vector pTF101.1 containing drought tolerance gene HVA1 from barley, and a selectable marker phosphinothricin (PPT) resistance (bar) gene. The explants were inoculated with A. tumefaciens (cell density OD650 at 0.7) at four different concentrations of acetosyringone (0, 100, 200, and 400 µM). The results showed that embryogenic calli from mature embryos showed higher regeneration and transformation than mature embryo halves and pieces. The integration of the transgene was confirmed by PCR amplification using primers specific to the bar gene, 2x35S promoter, and HVA1 gene. The transformation efficiency ranging from 0.33% to 2.33% was obtained in Amira variety using embryogenic calli and acetosyringone concentrations of 200 and 400 µM. The integration, as well as inheritance of the transgene, was confirmed by PCR amplification in T0 and T1 generations. This is the first report describing a genetic transformation of Moroccan durum wheat varieties via Agrobacterium tumefaciens. A seca é uma das principais restrições na produção de trigo da bacia do Mediterrâneo. A fim de superar este problema, a transformação genética é uma das opções para a melhoria da tolerância à seca. O objetivo deste estudo é investigar o efeito do tipo de explante e concentração de acetosinganona para o desenvolvimento eficiente do gene mediado por Agrobacterium em três variedades de trigo duro (Amria, Chaoui, Marouane). Embriões maduros foram inoculados com Agrobacterium tumefaciens, cepa EHA101, contendo o vetor binário pTF101.1 contendo o gene de tolerância à seca HVA1, e um gene de resistência (bar) marcado com a fosfinotricina (PPT). Os explantes foram inoculados com densidade de células de A. tumefaciens (OD650 a 0,7) em quatro diferentes concentrações de acetoseringona (0, 100, 200 e 400 µM). De acordo com os resultados de regeneração e seleção de plântulas, os calos embriogênicos apresentaram maior eficiência de regeneração e seleção do que as metades e partes de embriões maduros. A integração do transgene foi confirmada por amplificação por PCR utilizando primers específicos para os genes bar, 2x35S e HVA1. A eficiência de transformação variando de 0,33% a 2,33% foi obtida utilizando calos embriogênicos. A concentração de acetoseringona de 100 e 400 µM mostrou uma eficiência de transformação de 0,26%, que é maior do que o valor observado para 200 µM (0,04%). A integração bem como a herança do transgene foi confirmada por amplificação por PCR na geração T0 e T1. Este é o primeiro relatório descrevendo uma transformação genética do trigo duro marroquino via Agrobacterium tumefaciens.EDUFU2022-02-16info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://seer.ufu.br/index.php/biosciencejournal/article/view/5451310.14393/BJ-v38n0a2022-54513Bioscience Journal ; Vol. 38 (2022): Continuous Publication; e38007Bioscience Journal ; v. 38 (2022): Continuous Publication; e380071981-3163reponame:Bioscience journal (Online)instname:Universidade Federal de Uberlândia (UFU)instacron:UFUenghttps://seer.ufu.br/index.php/biosciencejournal/article/view/54513/33975Marrocos; Contemporary Copyright (c) 2022 Khadija Ahansal, Rabha Abdelwahd, Sripada Mahabala Udupa, Hanane Aadel, Fatima Gaboun, Mohammed Ibriz, Driss Iraqihttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessAhansal, KhadijaAbdelwahd, Rabha Udupa, Sripada Aadel, Hanane Gaboun, FatimaIbriz, MohammedIraqi, Driss2022-06-02T12:19:45Zoai:ojs.www.seer.ufu.br:article/54513Revistahttps://seer.ufu.br/index.php/biosciencejournalPUBhttps://seer.ufu.br/index.php/biosciencejournal/oaibiosciencej@ufu.br||1981-31631516-3725opendoar:2022-06-02T12:19:45Bioscience journal (Online) - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat |
title |
Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat |
spellingShingle |
Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat Ahansal, Khadija Agrobacterium tumefaciens Drought Tolerance Genetic Transformation HVA1 Gene Triticum turgidum L. Agricultural Sciences |
title_short |
Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat |
title_full |
Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat |
title_fullStr |
Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat |
title_full_unstemmed |
Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat |
title_sort |
Effect of type of mature embryo explants and acetosyringone on agrobacterium-mediated transformation of moroccan durum wheat |
author |
Ahansal, Khadija |
author_facet |
Ahansal, Khadija Abdelwahd, Rabha Udupa, Sripada Aadel, Hanane Gaboun, Fatima Ibriz, Mohammed Iraqi, Driss |
author_role |
author |
author2 |
Abdelwahd, Rabha Udupa, Sripada Aadel, Hanane Gaboun, Fatima Ibriz, Mohammed Iraqi, Driss |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Ahansal, Khadija Abdelwahd, Rabha Udupa, Sripada Aadel, Hanane Gaboun, Fatima Ibriz, Mohammed Iraqi, Driss |
dc.subject.por.fl_str_mv |
Agrobacterium tumefaciens Drought Tolerance Genetic Transformation HVA1 Gene Triticum turgidum L. Agricultural Sciences |
topic |
Agrobacterium tumefaciens Drought Tolerance Genetic Transformation HVA1 Gene Triticum turgidum L. Agricultural Sciences |
description |
Drought is one of the major constraints in durum wheat production in the Mediterranean Basin. In order to overcome this problem, the genetic transformation of durum wheat is one of the choices for improvement. However, the recalcitrance to Agrobacterium-mediated transformation in durum wheat (Triticum turgidum L.) is one of the factors limiting a successful genetic transformation. The aim of this study was to investigate the effect of explant type and acetosyringone concentration for the efficient Agrobacterium-mediated genetic transformation of three Moroccan durum wheat varieties (Amria, Chaoui, and Marouane). The mature embryos (intact, halved and pieces) were inoculated with Agrobacterium tumefaciens strain EHA101 harboring the binary vector pTF101.1 containing drought tolerance gene HVA1 from barley, and a selectable marker phosphinothricin (PPT) resistance (bar) gene. The explants were inoculated with A. tumefaciens (cell density OD650 at 0.7) at four different concentrations of acetosyringone (0, 100, 200, and 400 µM). The results showed that embryogenic calli from mature embryos showed higher regeneration and transformation than mature embryo halves and pieces. The integration of the transgene was confirmed by PCR amplification using primers specific to the bar gene, 2x35S promoter, and HVA1 gene. The transformation efficiency ranging from 0.33% to 2.33% was obtained in Amira variety using embryogenic calli and acetosyringone concentrations of 200 and 400 µM. The integration, as well as inheritance of the transgene, was confirmed by PCR amplification in T0 and T1 generations. This is the first report describing a genetic transformation of Moroccan durum wheat varieties via Agrobacterium tumefaciens. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-02-16 |
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/54513 10.14393/BJ-v38n0a2022-54513 |
url |
https://seer.ufu.br/index.php/biosciencejournal/article/view/54513 |
identifier_str_mv |
10.14393/BJ-v38n0a2022-54513 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://seer.ufu.br/index.php/biosciencejournal/article/view/54513/33975 |
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 |
Marrocos; Contemporary |
dc.publisher.none.fl_str_mv |
EDUFU |
publisher.none.fl_str_mv |
EDUFU |
dc.source.none.fl_str_mv |
Bioscience Journal ; Vol. 38 (2022): Continuous Publication; e38007 Bioscience Journal ; v. 38 (2022): Continuous Publication; e38007 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_ |
1797069082883784704 |