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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 13, No. 4, 2010
Bioline Code: ej10039
Full paper language: English
Document type: Report
Document available free of charge

Electronic Journal of Biotechnology, Vol. 13, No. 4, 2010

 en Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants
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. & Aguilar, José Juan Zúñiga

Abstract

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.

Keywords
cocultivation, GUS assay.

 
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