Plant Biotechnology
Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 13 No. 4, Issue of July 15, 2010
© 2010 by Pontificia Universidad Católica de Valparaíso -- Chile Received November 2, 2009 / Accepted January 29, 2010
DOI: 10.2225/vol13-issue4-fulltext-3
REPORT

Endogenous GUS-like activity in Capsicum chinense Jacq.

Laura Yesenia Solís-Ramos*#
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Calle 43, No.130, Chuburná de Hidalgo
C.P. 97200, Mérida, Yucatán, México
E-mail: laura.solisramos@ucr.ac.cr

Tomas González-Estrada¥
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Calle 43, No.130, Chuburná de Hidalgo
C.P. 97200, Mérida, Yucatán, México

Antonio Andrade-Torres
INBIOTECA
Universidad Veracruzana
Apdo. postal 250, C.P. 91000, Xalapa, Veracruz, México

Gregorio Godoy-Hernández
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Calle 43, No.130, Chuburná de Hidalgo
C.P. 97200, Mérida, Yucatán, México

Enrique Castaño de la Serna
Unidad de Bioquímica y Biología Molecular de Plantas
Centro de Investigación Científica de Yucatán
Calle 43, No.130, Chuburná de Hidalgo
C.P. 97200, Mérida, Yucatán, México

*Corresponding author

Financial support: Consejo Nacional de Ciencia y Tecnología (CONACYT), México and Dirección de Intercambio Académico de la Secretaría de Relaciones Exteriores from México.

Keywords: Agrobacterium tumefaciens, β-glucuronidase, Capsicum chinense, gene uidA.

Present address: #Escuela de Biología, Universidad de Costa Rica, P.O. Box 11501-2060, San Pedro, Costa Rica. ¥Consejo de Ciencia y Tecnología del Estado de Yucatán (CONCYTEY), Mérida, Yucatán, México.

Abbreviations:

DMSO: Dimethyl sulfoxide
DsRFP: Red fluorescent protein
EDTA: ethylene diamine tetraacetic acid
GUS: β-glucuronidase (gene uidA)
pH: hydrogen potential

Abstract   Full Text

The gene uidA, codes for β-glucuronidase, which is one of the reporters more frequently utilized in transgenic plants. However, this can only be use if the selected organism does not present endogenous GUS-like activity. In tissues of C. chinense we found a GUS-like activity showing different levels of intensity. Histochemical screening showed that endogenous GUS-like activity decreased, or reduced significantly, in almost all tissues with exception of stament, when phosphate buffer was adjusted to pH 8. Subsequently, C. chinense zygotic embryo explants were transient transformed with Agrobacterium tumefaciens LBA4404 (pCAMBIA2301) and plantlets regenerated were histochemically stained in phosphate buffer pH 8. Observations of incubated tissues of C. chinense regenerants showed blue staining, suggesting expression of uidA. Incubated tissues of non-transformed regenerants did not show blue staining in phosphate buffer pH 8. The results show that for transformation experiments of C. chinense with uidA gene, pH 8 is recommended for histochemical staining.

Supported by UNESCO / MIRCEN network