Published

2016-09-01

Molecular characterization and Freedom to Operate analysis of maize hybrids from genetically modified and Colombian varieties

Caracterización molecular y análisis de libertad de operación de híbridos de maíz de variedades colombianas y genéticamente modificadas

DOI:

https://doi.org/10.15446/agron.colomb.v34n3.60350

Keywords:

crop improvement, PCR, pest resistance. (en)
mejora de cultivos, PCR, resistencia a plagas (es)

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Authors

  • Jenny Jiménez-Barreto Universidad Nacional de Colombia - Sede Bogotá
  • Alejandro Chaparro Giraldo Universidad Nacional de Colombia - Sede Bogotá
  • Julián Mora-Oberlaender Universidad Nacional de Colombia - Sede Bogotá
  • José Ever Vargas-Sánchez Centro de Investigación y Transferencia de Tecnología de la Cadena Agroalimentaria de los Cereales y las Leguminosas (CENICEL)

Spodoptera frugiperda is one of the most common pests in maize crops, causing important losses in Colombia. The development of insect resistant crops is an alternative to conventional pest control methods. HERCULEX® I is a transgenic maize line with resistance to lepidopteran insects and tolerance to phosphinothricin herbicides, conferred by genes cry1F and pat, respectively. In Colombia, the National Federation of Cereal and Legume Growers (FENALCE) has carried out mendelian crosses between Colombian maize genotypes and HERCU- LEX® I, with the aim of integrating the genetic elements of the transgenic line into national varieties. Three hybrids were obtained which potentially carry such constructs. In the present study, the hybrid lines and their parental lines were characterized at the molecular level. Leaf tissue was tested for presence or absence of the genes cry1F and pat and their expression as mRNA and respective proteins Cry1F and Pat. Results show that the three hybrids indeed carry the HERCULEX® I constructs. RT-PCR and ELISA analysis showed transcription of the genes and presence of the proteins. An initial approach to the Freedom to Operate analysis was carried out for HERCU- LEX® I in Colombia.

Spodoptera frugiperda es una de las plagas más comunes del maíz, que causa pérdidas económicas importantes en Colom bia. Una alternativa a los métodos convencionales de control de plagas, es la producción de cultivos con resistencia a insectos. HERCULEX®I es una línea de maíz transgénico caracterizada por ser resistente a lepidópteros y tolerante a herbicidas tipo fosfinotricina, características provistas por los genes crylF y pat. Con el objetivo de integrar estas características en maíz colombiano, la Federación Nacional de Cultivadores de Cere ales y Oleaginosas (FENALCE) ha desarrollado experimentos de cruzamientos mendelianos entre genotipos colombianos de maíz y HERCULEX® I. De estos experimentos se obtuvi eron tres híbridos que potencialmente podrían contener en su genoma el constructo de HERCULEX® I. En este trabajo se caracterizaron molecularmente dichos híbridos y sus respec tivos parentales. El análisis se realizó mediante la detección en hojas de presencia/ausencia de los genes crylF y pat, pruebas de expresión en ARN y detección de las proteínas CrylF y Pat. Los resultados indican que los híbridos contienen en su genoma elementos característicos de HERCULEX® I. Mediante RT-PCR y ELISA se determinó el transcripto primario de los genes y la proteína de interés. Adicionalmente, se realizó una aproximación inicial al análisis de libertad de operación de HERCULEX® I para Colombia.

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Molecular characterization and Freedom to Operate analysis of maize hybrids from genetically modified and Colombian varieties. (2016). Agronomía Colombiana, 34(3), 309-316. https://doi.org/10.15446/agron.colomb.v34n3.60350

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