Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants

Most of the pepper species of the genus Capsicum have been recalcitrant to efficient Agrobacterium tumefaciens-mediated stable or transient, genetic transformation. In the present work, we optimized a protocol for transient transformation of the Habanero pepper (Capsicum chinense Jacq.) through the standardization of several experimental factors. These included the age of the plants, the temperature, the length of co-cultivation, the application of a negative (vacuum) and/or a positive (infiltration) pressure, along with micro injection, the use of acetosyringone during the bacterial culturing, and modification of the pH during the GUS assay to eliminate the endogenous β-glucuronidase activity. The standardized protocol, which yielded nearly 55% fully transformed leaf explants, was used to successfully mobilize two empty binary vectors (pCAMBIA2301 and pCAMex), as well as the C. chinense cDNAs encoding the pathogenesis-related protein 10 and esterase, respectively.

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Main Authors: GUADALUPE FABIOLA ARCOS ORTEGA, RAFAEL ANTONIO CHAN KUUK, WILMA ARACELY GONZALEZ KANTUN, RAMON ARMANDO SOUZA PERERA, YUMI ELENA NAKAZAWA UEJI, ELIDE AVILES BERZUNZA, GREGORIO DEL CARMEN GODOY HERNANDEZ, Michael Lawton, JOSE JUAN ZUÑIGA AGUILAR
Format: info:eu-repo/semantics/article biblioteca
Language:eng
Subjects:info:eu-repo/classification/Autores/COCULTIVATION, info:eu-repo/classification/Autores/GUS ASSAY, info:eu-repo/classification/cti/2,
Online Access:http://cicy.repositorioinstitucional.mx/jspui/handle/1003/1318
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spelling dig-cicy-1003-13182018-10-25T16:41:48Z Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants GUADALUPE FABIOLA ARCOS ORTEGA RAFAEL ANTONIO CHAN KUUK WILMA ARACELY GONZALEZ KANTUN RAMON ARMANDO SOUZA PERERA YUMI ELENA NAKAZAWA UEJI ELIDE AVILES BERZUNZA GREGORIO DEL CARMEN GODOY HERNANDEZ Michael Lawton JOSE JUAN ZUÑIGA AGUILAR 2010 info:eu-repo/semantics/article Most of the pepper species of the genus Capsicum have been recalcitrant to efficient Agrobacterium tumefaciens-mediated stable or transient, genetic transformation. In the present work, we optimized a protocol for transient transformation of the Habanero pepper (Capsicum chinense Jacq.) through the standardization of several experimental factors. These included the age of the plants, the temperature, the length of co-cultivation, the application of a negative (vacuum) and/or a positive (infiltration) pressure, along with micro injection, the use of acetosyringone during the bacterial culturing, and modification of the pH during the GUS assay to eliminate the endogenous β-glucuronidase activity. The standardized protocol, which yielded nearly 55% fully transformed leaf explants, was used to successfully mobilize two empty binary vectors (pCAMBIA2301 and pCAMex), as well as the C. chinense cDNAs encoding the pathogenesis-related protein 10 and esterase, respectively. info:eu-repo/classification/Autores/COCULTIVATION info:eu-repo/classification/Autores/GUS ASSAY info:eu-repo/classification/cti/2 info:eu-repo/classification/cti/2 Electronic Journal of Biotechnology, 13(4), 7-8, 2010. http://cicy.repositorioinstitucional.mx/jspui/handle/1003/1318 info:eu-repo/semantics/datasetDOI/10.2225/vol13-issue4-fulltext-10 info:eu-repo/semantics/openAccess eng citation:Arcos-Ortega, G. F., Chan-Kuuk, R. A., González-Kantún, W. A., Souza-Perera, R., Nakazawa-Ueji, Y. E., Avilés-Berzunza, E., ... & Zúñiga Aguilar, J. J. (2010). Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants. Electronic Journal of Biotechnology, 13(4), 7-8. http://creativecommons.org/licenses/by-nc-nd/4.0 info:eu-repo/semantics/publishedVersion application/pdf
institution CICY
collection DSpace
country México
countrycode MX
component Bibliográfico
access En linea
databasecode dig-cicy
tag biblioteca
region America del Norte
libraryname Biblioteca del CICY
language eng
topic info:eu-repo/classification/Autores/COCULTIVATION
info:eu-repo/classification/Autores/GUS ASSAY
info:eu-repo/classification/cti/2
info:eu-repo/classification/cti/2
info:eu-repo/classification/Autores/COCULTIVATION
info:eu-repo/classification/Autores/GUS ASSAY
info:eu-repo/classification/cti/2
info:eu-repo/classification/cti/2
spellingShingle info:eu-repo/classification/Autores/COCULTIVATION
info:eu-repo/classification/Autores/GUS ASSAY
info:eu-repo/classification/cti/2
info:eu-repo/classification/cti/2
info:eu-repo/classification/Autores/COCULTIVATION
info:eu-repo/classification/Autores/GUS ASSAY
info:eu-repo/classification/cti/2
info:eu-repo/classification/cti/2
GUADALUPE FABIOLA ARCOS ORTEGA
RAFAEL ANTONIO CHAN KUUK
WILMA ARACELY GONZALEZ KANTUN
RAMON ARMANDO SOUZA PERERA
YUMI ELENA NAKAZAWA UEJI
ELIDE AVILES BERZUNZA
GREGORIO DEL CARMEN GODOY HERNANDEZ
Michael Lawton
JOSE JUAN ZUÑIGA AGUILAR
Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants
description Most of the pepper species of the genus Capsicum have been recalcitrant to efficient Agrobacterium tumefaciens-mediated stable or transient, genetic transformation. In the present work, we optimized a protocol for transient transformation of the Habanero pepper (Capsicum chinense Jacq.) through the standardization of several experimental factors. These included the age of the plants, the temperature, the length of co-cultivation, the application of a negative (vacuum) and/or a positive (infiltration) pressure, along with micro injection, the use of acetosyringone during the bacterial culturing, and modification of the pH during the GUS assay to eliminate the endogenous β-glucuronidase activity. The standardized protocol, which yielded nearly 55% fully transformed leaf explants, was used to successfully mobilize two empty binary vectors (pCAMBIA2301 and pCAMex), as well as the C. chinense cDNAs encoding the pathogenesis-related protein 10 and esterase, respectively.
format info:eu-repo/semantics/article
topic_facet info:eu-repo/classification/Autores/COCULTIVATION
info:eu-repo/classification/Autores/GUS ASSAY
info:eu-repo/classification/cti/2
info:eu-repo/classification/cti/2
author GUADALUPE FABIOLA ARCOS ORTEGA
RAFAEL ANTONIO CHAN KUUK
WILMA ARACELY GONZALEZ KANTUN
RAMON ARMANDO SOUZA PERERA
YUMI ELENA NAKAZAWA UEJI
ELIDE AVILES BERZUNZA
GREGORIO DEL CARMEN GODOY HERNANDEZ
Michael Lawton
JOSE JUAN ZUÑIGA AGUILAR
author_facet GUADALUPE FABIOLA ARCOS ORTEGA
RAFAEL ANTONIO CHAN KUUK
WILMA ARACELY GONZALEZ KANTUN
RAMON ARMANDO SOUZA PERERA
YUMI ELENA NAKAZAWA UEJI
ELIDE AVILES BERZUNZA
GREGORIO DEL CARMEN GODOY HERNANDEZ
Michael Lawton
JOSE JUAN ZUÑIGA AGUILAR
author_sort GUADALUPE FABIOLA ARCOS ORTEGA
title Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants
title_short Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants
title_full Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants
title_fullStr Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants
title_full_unstemmed Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants
title_sort agrobacterium tumefaciens-transient genetic transformation of habanero pepper (capsicum chinense jacq.) leaf explants
url http://cicy.repositorioinstitucional.mx/jspui/handle/1003/1318
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