Improving crops genome through genetic engineering of the key metabolic pathways
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Acta Scientiarum Biological Sciences |
Texto Completo: | http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/52272 |
Resumo: | Soybean loss due to pests and pathogens is a serious problem worldwide. Soybean producers have few options to manage diseases caused by general pathogens where major genes for full resistance have not been discovered. The innate defense of soybean plants could be enhanced by improving content and composition of lignin by genetic engineering of the phenylpropanoid pathway. We used a novel technique of germ-line genetic transformation of soybean plants via natural pollen tubes as vectors. This technique uses Agrobacterium tumefaciens to mediate transfer of genes of interest to the zygote to introduce the key lignification genes (PtMYB4, PAL5, F5H, CAD1) into soybean genome. We observed 5.6% average transformation efficiency in the first generation of transgenic plants and in the second generation the presence of the transgene constructs was confirmed in more than 50% (for CsVMV/PtMYB4sens, 35SVTM/PAL5, C4H/F5H, CsVMV/CAD1 constructs) transgenic soybean lines. We confirmed the expression of the introduced genes at transcriptional level using RT-PCR and Northern blot. Functional analysis using lignin content determination and the activity of PAL5 and CAD1 enzymes demonstrated that the transgenes perform their function in planta. The proposed technique is effective and inexpensive and can be used to create novel stress and disease resistant soybean genotypes. |
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Improving crops genome through genetic engineering of the key metabolic pathwaysImproving crops genome through genetic engineering of the key metabolic pathwaysgerm-line genetic transformation; lignification; soybean; biotic stress resistance.germ-line genetic transformation; lignification; soybean; biotic stress resistance.Soybean loss due to pests and pathogens is a serious problem worldwide. Soybean producers have few options to manage diseases caused by general pathogens where major genes for full resistance have not been discovered. The innate defense of soybean plants could be enhanced by improving content and composition of lignin by genetic engineering of the phenylpropanoid pathway. We used a novel technique of germ-line genetic transformation of soybean plants via natural pollen tubes as vectors. This technique uses Agrobacterium tumefaciens to mediate transfer of genes of interest to the zygote to introduce the key lignification genes (PtMYB4, PAL5, F5H, CAD1) into soybean genome. We observed 5.6% average transformation efficiency in the first generation of transgenic plants and in the second generation the presence of the transgene constructs was confirmed in more than 50% (for CsVMV/PtMYB4sens, 35SVTM/PAL5, C4H/F5H, CsVMV/CAD1 constructs) transgenic soybean lines. We confirmed the expression of the introduced genes at transcriptional level using RT-PCR and Northern blot. Functional analysis using lignin content determination and the activity of PAL5 and CAD1 enzymes demonstrated that the transgenes perform their function in planta. The proposed technique is effective and inexpensive and can be used to create novel stress and disease resistant soybean genotypes.Soybean loss due to pests and pathogens is a serious problem worldwide. Soybean producers have few options to manage diseases caused by general pathogens where major genes for full resistance have not been discovered. The innate defense of soybean plants could be enhanced by improving content and composition of lignin by genetic engineering of the phenylpropanoid pathway. We used a novel technique of germ-line genetic transformation of soybean plants via natural pollen tubes as vectors. This technique uses Agrobacterium tumefaciens to mediate transfer of genes of interest to the zygote to introduce the key lignification genes (PtMYB4, PAL5, F5H, CAD1) into soybean genome. We observed 5.6% average transformation efficiency in the first generation of transgenic plants and in the second generation the presence of the transgene constructs was confirmed in more than 50% (for CsVMV/PtMYB4sens, 35SVTM/PAL5, C4H/F5H, CsVMV/CAD1 constructs) transgenic soybean lines. We confirmed the expression of the introduced genes at transcriptional level using RT-PCR and Northern blot. Functional analysis using lignin content determination and the activity of PAL5 and CAD1 enzymes demonstrated that the transgenes perform their function in planta. The proposed technique is effective and inexpensive and can be used to create novel stress and disease resistant soybean genotypes.Universidade Estadual De Maringá2020-08-24info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/5227210.4025/actascibiolsci.v42i1.52272Acta Scientiarum. Biological Sciences; Vol 42 (2020): Publicação contínua; e52272Acta Scientiarum. Biological Sciences; v. 42 (2020): Publicação contínua; e522721807-863X1679-9283reponame:Acta Scientiarum Biological Sciencesinstname:Universidade Estadual de Maringá (UEM)instacron:UEMenghttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/52272/751375150588Copyright (c) 2020 Acta Scientiarum. Biological Scienceshttp://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessKershanskaya, Olga Ivanovna Nelidova, Darya Sergeevna Yessenbaeva , Gulvira Lemesovna Nelidov, Sergey Nikolaevich 2020-11-16T16:25:43Zoai:periodicos.uem.br/ojs:article/52272Revistahttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSciPUBhttp://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/oai||actabiol@uem.br1807-863X1679-9283opendoar:2020-11-16T16:25:43Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM)false |
dc.title.none.fl_str_mv |
Improving crops genome through genetic engineering of the key metabolic pathways Improving crops genome through genetic engineering of the key metabolic pathways |
title |
Improving crops genome through genetic engineering of the key metabolic pathways |
spellingShingle |
Improving crops genome through genetic engineering of the key metabolic pathways Kershanskaya, Olga Ivanovna germ-line genetic transformation; lignification; soybean; biotic stress resistance. germ-line genetic transformation; lignification; soybean; biotic stress resistance. |
title_short |
Improving crops genome through genetic engineering of the key metabolic pathways |
title_full |
Improving crops genome through genetic engineering of the key metabolic pathways |
title_fullStr |
Improving crops genome through genetic engineering of the key metabolic pathways |
title_full_unstemmed |
Improving crops genome through genetic engineering of the key metabolic pathways |
title_sort |
Improving crops genome through genetic engineering of the key metabolic pathways |
author |
Kershanskaya, Olga Ivanovna |
author_facet |
Kershanskaya, Olga Ivanovna Nelidova, Darya Sergeevna Yessenbaeva , Gulvira Lemesovna Nelidov, Sergey Nikolaevich |
author_role |
author |
author2 |
Nelidova, Darya Sergeevna Yessenbaeva , Gulvira Lemesovna Nelidov, Sergey Nikolaevich |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Kershanskaya, Olga Ivanovna Nelidova, Darya Sergeevna Yessenbaeva , Gulvira Lemesovna Nelidov, Sergey Nikolaevich |
dc.subject.por.fl_str_mv |
germ-line genetic transformation; lignification; soybean; biotic stress resistance. germ-line genetic transformation; lignification; soybean; biotic stress resistance. |
topic |
germ-line genetic transformation; lignification; soybean; biotic stress resistance. germ-line genetic transformation; lignification; soybean; biotic stress resistance. |
description |
Soybean loss due to pests and pathogens is a serious problem worldwide. Soybean producers have few options to manage diseases caused by general pathogens where major genes for full resistance have not been discovered. The innate defense of soybean plants could be enhanced by improving content and composition of lignin by genetic engineering of the phenylpropanoid pathway. We used a novel technique of germ-line genetic transformation of soybean plants via natural pollen tubes as vectors. This technique uses Agrobacterium tumefaciens to mediate transfer of genes of interest to the zygote to introduce the key lignification genes (PtMYB4, PAL5, F5H, CAD1) into soybean genome. We observed 5.6% average transformation efficiency in the first generation of transgenic plants and in the second generation the presence of the transgene constructs was confirmed in more than 50% (for CsVMV/PtMYB4sens, 35SVTM/PAL5, C4H/F5H, CsVMV/CAD1 constructs) transgenic soybean lines. We confirmed the expression of the introduced genes at transcriptional level using RT-PCR and Northern blot. Functional analysis using lignin content determination and the activity of PAL5 and CAD1 enzymes demonstrated that the transgenes perform their function in planta. The proposed technique is effective and inexpensive and can be used to create novel stress and disease resistant soybean genotypes. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-08-24 |
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 |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/52272 10.4025/actascibiolsci.v42i1.52272 |
url |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/52272 |
identifier_str_mv |
10.4025/actascibiolsci.v42i1.52272 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
http://www.periodicos.uem.br/ojs/index.php/ActaSciBiolSci/article/view/52272/751375150588 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2020 Acta Scientiarum. Biological Sciences http://creativecommons.org/licenses/by/4.0 info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2020 Acta Scientiarum. Biological Sciences http://creativecommons.org/licenses/by/4.0 |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
publisher.none.fl_str_mv |
Universidade Estadual De Maringá |
dc.source.none.fl_str_mv |
Acta Scientiarum. Biological Sciences; Vol 42 (2020): Publicação contínua; e52272 Acta Scientiarum. Biological Sciences; v. 42 (2020): Publicação contínua; e52272 1807-863X 1679-9283 reponame:Acta Scientiarum Biological Sciences instname:Universidade Estadual de Maringá (UEM) instacron:UEM |
instname_str |
Universidade Estadual de Maringá (UEM) |
instacron_str |
UEM |
institution |
UEM |
reponame_str |
Acta Scientiarum Biological Sciences |
collection |
Acta Scientiarum Biological Sciences |
repository.name.fl_str_mv |
Acta Scientiarum Biological Sciences - Universidade Estadual de Maringá (UEM) |
repository.mail.fl_str_mv |
||actabiol@uem.br |
_version_ |
1799317391043198976 |