TY - JOUR
T1 - Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants
AU - Arcos-Ortega, Guadalupe Fabiola
AU - Chan-Kuuk, Rafael Antonio
AU - González-Kantún, Wilma Aracely
AU - Souza-Perera, Ramón
AU - Nakazawa-Ueji, Yumi Elena
AU - Avilés-Berzunza, Elidé
AU - Godoy-Hernández, Gregorio
AU - Lawton, Michael A.
AU - Aguilar, José Juan Zúñiga
PY - 2010
Y1 - 2010
N2 - 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 pathogenesisrelated protein 10 and esterase, respectively.
AB - 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 pathogenesisrelated protein 10 and esterase, respectively.
KW - Cocultivation
KW - GUS assay
UR - http://www.scopus.com/inward/record.url?scp=79960818623&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79960818623&partnerID=8YFLogxK
U2 - 10.2225/vol13-issue4-fulltext-10
DO - 10.2225/vol13-issue4-fulltext-10
M3 - Article
AN - SCOPUS:79960818623
SN - 0717-3458
VL - 13
JO - Electronic Journal of Biotechnology
JF - Electronic Journal of Biotechnology
IS - 4
ER -