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

Guadalupe Fabiola Arcos-Ortega, Rafael Antonio Chan-Kuuk, Wilma Aracely González-Kantún, Ramón Souza-Perera, Yumi Elena Nakazawa-Ueji, Elidé Avilés-Berzunza, Gregorio Godoy-Hernández, Michael Lawton, José Juan Zúñiga Aguilar

Research output: Contribution to journalArticle

11 Citations (Scopus)

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 pathogenesisrelated protein 10 and esterase, respectively.

Original languageEnglish (US)
JournalElectronic Journal of Biotechnology
Volume13
Issue number4
DOIs
StatePublished - Dec 1 2010

Fingerprint

Genetic Transformation
Capsicum
Agrobacterium tumefaciens
Carboxylesterase
Glucuronidase
Vacuum
Complementary DNA
Pressure
Injections
Temperature
Proteins
acetosyringone

All Science Journal Classification (ASJC) codes

  • Applied Microbiology and Biotechnology
  • Biotechnology

Keywords

  • Cocultivation
  • GUS assay

Cite this

Arcos-Ortega, G. F., Chan-Kuuk, R. A., González-Kantún, W. A., Souza-Perera, R., Nakazawa-Ueji, Y. E., Avilés-Berzunza, E., ... Aguilar, J. J. Z. (2010). Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants. Electronic Journal of Biotechnology, 13(4). https://doi.org/10.2225/vol13-issue4-fulltext-10
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 ; Aguilar, José Juan Zúñiga. / Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants. In: Electronic Journal of Biotechnology. 2010 ; Vol. 13, No. 4.
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Arcos-Ortega, GF, Chan-Kuuk, RA, González-Kantún, WA, Souza-Perera, R, Nakazawa-Ueji, YE, Avilés-Berzunza, E, Godoy-Hernández, G, Lawton, M & Aguilar, JJZ 2010, 'Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants', Electronic Journal of Biotechnology, vol. 13, no. 4. https://doi.org/10.2225/vol13-issue4-fulltext-10

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; Aguilar, José Juan Zúñiga.

In: Electronic Journal of Biotechnology, Vol. 13, No. 4, 01.12.2010.

Research output: Contribution to journalArticle

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

AU - Aguilar, José Juan Zúñiga

PY - 2010/12/1

Y1 - 2010/12/1

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

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M3 - Article

VL - 13

JO - Electronic Journal of Biotechnology

JF - Electronic Journal of Biotechnology

SN - 0717-3458

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Arcos-Ortega GF, Chan-Kuuk RA, González-Kantún WA, Souza-Perera R, Nakazawa-Ueji YE, Avilés-Berzunza E et al. Agrobacterium tumefaciens-transient genetic transformation of Habanero pepper (Capsicum chinense Jacq.) leaf explants. Electronic Journal of Biotechnology. 2010 Dec 1;13(4). https://doi.org/10.2225/vol13-issue4-fulltext-10