Transient expression of a green fluorescent protein in tobacco and maize chloroplast

Electronic Journal of Biotechnology. 2020;45:1-9

 

Journal Homepage

Journal Title: Electronic Journal of Biotechnology

ISSN: 0717-3458 (Online)

Publisher: Elsevier

Society/Institution: Pontificia Universidad Católica de Valparaíso

LCC Subject Category: Technology: Chemical technology: Biotechnology | Science: Biology (General)

Country of publisher: Netherlands

Language of fulltext: English

Full-text formats available: PDF, HTML

 

AUTHORS


Sigifredo Arévalo-Gallegos (Laboratorio de Biotecnología I, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito Universitarios s/n, Nuevo Campus Universitario, C.P. 31125, Chihuahua, Chihuahua, México)

Hugo Varela-Rodríguez (Laboratorio de Biotecnología I, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito Universitarios s/n, Nuevo Campus Universitario, C.P. 31125, Chihuahua, Chihuahua, México)

Héctor Lugo-Aguilar (Laboratorio de Biotecnología I, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito Universitarios s/n, Nuevo Campus Universitario, C.P. 31125, Chihuahua, Chihuahua, México)

Tania S. Siqueiros-Cendón (Laboratorio de Biotecnología I, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito Universitarios s/n, Nuevo Campus Universitario, C.P. 31125, Chihuahua, Chihuahua, México)

Blanca F. Iglesias-Figueroa (Laboratorio de Biotecnología I, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito Universitarios s/n, Nuevo Campus Universitario, C.P. 31125, Chihuahua, Chihuahua, México)

Edward A. Espinoza-Sánchez (Laboratorio de Biotecnología I, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito Universitarios s/n, Nuevo Campus Universitario, C.P. 31125, Chihuahua, Chihuahua, México)

Gerardo A. Aguado-Santacruz (Instituto Tecnológico de Roque, Carretera Celaya-Juventino Rosas Km. 8, C.P. 38110, Celaya, Guanajuato, México)

Quintín Rascón-Cruz (Laboratorio de Biotecnología I, Facultad de Ciencias Químicas, Universidad Autónoma de Chihuahua, Circuito Universitarios s/n, Nuevo Campus Universitario, C.P. 31125, Chihuahua, Chihuahua, México; Corresponding author.)

EDITORIAL INFORMATION

Blind peer review

Editorial Board

Instructions for authors

Time From Submission to Publication: 10 weeks

 

Abstract | Full Text

Background: Maize is one of the most important crops worldwide and has been a target of nuclear-based transformation biotechnology to improve it and satisfy the food demand of the ever-growing global population. However, the maize plastid transformation has not been accomplished due to the recalcitrant condition of the crop. Results: In this study, we constructed two different vectors with homologous recombination sequences from maize (Zea mays var. LPC13) and grass (Bouteloua gracilis var. ex Steud) (pZmcpGFP and pBgcpGFP, respectively). Both vectors were designed to integrate into rrn23S/rrn16S from an inverted repeat region in the chloroplast genome. Moreover, the vector had the mgfp5 gene driven by Prrn, a leader sequence of the atpB gene and a terminator sequence from the rbcL gene. Also, constructs have an hph gene as a selection marker gene driven by Prrn, a leader sequence from rbcL gene and a terminator sequence from the rbcL gene. Explants of maize, tobacco and Escherichia coli cells were transformed with both vectors to evaluate the transitory expression–an exhibition of green and red fluorescent light under epifluorescence microscopy. These results showed that both vectors were expressed; the reporter gene in all three organisms confirmed the capacity of the vectors to express genes in the cell compartments. Conclusions: This paper is the first report of transient expression of GFP in maize embryos and offers new information for genetically improving recalcitrant crops; it also opens new possibilities for the improvement in maize chloroplast transformation with these vectors.How to cite: Arévalo-Gallegos S, Varela-Rodríguez H, Lugo-Aguilar H, et al. Transient expression of a green fluorescent protein in tobacco and maize chloroplast. Electron J Biotechnol 2020;44. https://doi.org/10.1016/j.ejbt.2020.01.008