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...
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Pontificia Universidad Católica de Valparaíso
2010
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oai:scielo:S0717-345820100004000072010-08-04Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explantsArcos-Ortega,Guadalupe FabiolaChan-Kuuk,Rafael AntonioGonzález-Kantún,Wilma AracelySouza-Perera,RamónNakazawa-Ueji,Yumi ElenaAvilés-Berzunza,ElidéGodoy-Hernández,GregorioLawton,Michael AZúñiga Aguilar,José Juan cocultivation GUS assay 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/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.4 20102010-07-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000400007en |
| institution |
Scielo Chile |
| collection |
Scielo Chile |
| language |
English |
| topic |
cocultivation GUS assay |
| spellingShingle |
cocultivation GUS assay Arcos-Ortega,Guadalupe Fabiola Chan-Kuuk,Rafael Antonio González-Kantún,Wilma Aracely Souza-Perera,Ramón Nakazawa-Ueji,Yumi Elena Avilés-Berzunza,Elidé Godoy-Hernández,Gregorio Lawton,Michael A Zúñiga Aguilar,José Juan 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. |
| author |
Arcos-Ortega,Guadalupe Fabiola Chan-Kuuk,Rafael Antonio González-Kantún,Wilma Aracely Souza-Perera,Ramón Nakazawa-Ueji,Yumi Elena Avilés-Berzunza,Elidé Godoy-Hernández,Gregorio Lawton,Michael A Zúñiga Aguilar,José Juan |
| author_facet |
Arcos-Ortega,Guadalupe Fabiola Chan-Kuuk,Rafael Antonio González-Kantún,Wilma Aracely Souza-Perera,Ramón Nakazawa-Ueji,Yumi Elena Avilés-Berzunza,Elidé Godoy-Hernández,Gregorio Lawton,Michael A Zúñiga Aguilar,José Juan |
| author_sort |
Arcos-Ortega,Guadalupe Fabiola |
| 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 |
| publisher |
Pontificia Universidad Católica de Valparaíso |
| publishDate |
2010 |
| url |
http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000400007 |
| work_keys_str_mv |
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| _version_ |
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