Transformation of the phytopathogenic fungus Sclerotium cepivorum Berk using protoplasts fusion

Authors

  • Gerardo Acosta-García Departamento de Ingeniería Bioquímica, Instituto Tecnológico de Celaya, Av. Tecnológico y A. García Cubas S/N, Celaya, Guanajuato, México, C. P. 38010. Tel. 01 461 6117575, Ext. 324 y Fax, Ext. 402
  • Miguel Ángel Pantoja-Hernández Departamento de Ingeniería Bioquímica, Instituto Tecnológico de Celaya, Av. Tecnológico y A. García Cubas S/N, Celaya, Guanajuato, México, C. P. 38010. Tel. 01 461 6117575, Ext. 324 y Fax, Ext. 402
  • Claudia Ivonne Muñoz-Sánchez Departamento de Ingeniería Bioquímica, Instituto Tecnológico de Celaya, Av. Tecnológico y A. García Cubas S/N, Celaya, Guanajuato, México, C. P. 38010. Tel. 01 461 6117575, Ext. 324 y Fax, Ext. 402
  • Cristina Pérez-Pérez Departamento de Ingeniería Bioquímica, Instituto Tecnológico de Celaya, Av. Tecnológico y A. García Cubas S/N, Celaya, Guanajuato, México, C. P. 38010. Tel. 01 461 6117575, Ext. 324 y Fax, Ext. 402
  • Ramón Gerardo Guevara-González Ingeniería de Biosistemas. Facultad de Ingeniería. Universidad Autónoma de Querétaro. C. P 76010. Santiago de Querétaro, Querétaro. México
  • Irineo Torres-Pacheco Ingeniería de Biosistemas. Facultad de Ingeniería. Universidad Autónoma de Querétaro. C. P 76010. Santiago de Querétaro, Querétaro. México
  • Felipe Delgadillo-Sánchez Unidad de Biotecnología del Bajío. Campo Experimental Bajío- INIFAP. Carretera Celaya-San Miguel de Allende km 6.5, Celaya, Guanajuato, México
  • Mario Martín González-Chavira Unidad de Biotecnología del Bajío. Campo Experimental Bajío- INIFAP. Carretera Celaya-San Miguel de Allende km 6.5, Celaya, Guanajuato, México
  • Lorenzo Guevara-Olvera Departamento de Ingeniería Bioquímica, Instituto Tecnológico de Celaya, Av. Tecnológico y A. García Cubas S/N, Celaya, Guanajuato, México, C. P. 38010. Tel. 01 461 6117575, Ext. 324 y Fax, Ext. 402

DOI:

https://doi.org/10.29312/remexca.v3i7.1341

Keywords:

Allium sativum, hph gene of Escherichia coli, Sclerotium cepivorum, expression signals of Aspergillus nidulans, white rot

Abstract

White rot, a disease of most concern to the producers of garlic (Allium sativum), is caused by the phytopathogenic fungus Sclerotium cepivorum Berk. In order to develop procedures of molecular biology to the fungus, in this paper we describe a transformation method using fusion of protoplasts in the presence of polyethylene glycol 3350 (PEG). We used the plasmid pAN7-1 which confers resistance to hygromycin B, which contains the hygromycin phosphotransferase gene B (hph) from Escherichia coli under the glyceraldehyde phosphate dehydrogenase promoter (GPDA) and the anthranilate synthase terminator (trpC) of the filamentous fungus Aspergillus nidulans. The recovered transformants in the regeneration medium potato dextrose agar-sorbitol (PDA-S) in the presence of 200 μgmL -1 of hygromycin B were analyzed by amplifying a fragment of the bacterial gene hph by PCR with specific oligonucleotides: Hig 1, Sentido 5' GAC CTG CTG AGG TCC CTC 3 ́ and Hig 2, antisentido 5' GCC CTC GGA CGA GTG CTG 3 ́. Obtaining 12-18 frequencies of transformants of S. cepivorum per μg of vector DNA. The results described in this paper will contribute to the molecular characterization of the genes involved in the phenomenon of pathogenesis of S. cepivorum on garlic.

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Vol. 3, Núm. 7

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Published

2018-06-19

How to Cite

Acosta-García Gerardo, Pantoja-Hernández Miguel Ángel, Muñoz-Sánchez Claudia Ivonne, Pérez-Pérez Cristina, Guevara-González Ramón Gerardo, Irineo Torres-Pacheco, Delgadillo-Sánchez Felipe, González-Chavira Mario Martín, and Lorenzo Guevara-Olvera. 2018. “Transformation of the Phytopathogenic Fungus Sclerotium Cepivorum Berk Using Protoplasts Fusion”. Revista Mexicana De Ciencias Agrícolas 3 (7). México, ME:1333-45. https://doi.org/10.29312/remexca.v3i7.1341.

Issue

Vol. 3 No. 7 (2012)

Section

Articles

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