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

Guadalupe Fabiola Arcos-Ortega
Centro de Investigaciones Biológicas del Noroeste
Col. PlayaPalo de Santa Rita
La Paz, B.C.S., México 

Rafael Antonio Chan-Kuuk
Instituto Tecnológico Superior de Calkiní
Calkiní, Campeche, México 

Wilma Aracely González-Kantún
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Col. Chuburná de Hidalgo
Mérida, Yucatán, México 

Ramón Souza-Pereira
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Col. Chuburná de Hidalgo
Mérida, Yucatán, México 

Yumi Elena Nakazawa-Ueji
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Col. Chuburná de Hidalgo
Mérida, Yucatán, México 

Elidé Avilés-Berzunza
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Col. Chuburná de Hidalgo
Mérida, Yucatán, México 

Gregorio Godoy-Hernández
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Col. Chuburná de Hidalgo
Mérida, Yucatán, México 

Michael A. Lawton
Biotechnological Center for the Agriculture and the Environment
Cook College, Rutgers
The State University of New Jersey
New Brunswick, NJ, USA 

José Juan Zúñiga Aguilar*
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Col. Chuburná de Hidalgo
Mérida, Yucatán, México
E-mail: zuniga_cicy@hotmail.com

*Corresponding author

Financial support: This work was funded by the Consejo Nacional de Ciencia y Tecnología (Project P48831).

Keywords: 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.