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Strain Identification of Spodoptera frugiperda (Lepidoptera: Noctuidae) Insects and Cell Line: PCR-RFLP of Cytochrome Oxidase C Subunit I Gene

Hazel C. Levy, Alejandra Garcia-Maruniak and James E. Maruniak
The Florida Entomologist
Vol. 85, No. 1 (Mar., 2002), pp. 186-190
Stable URL: http://www.jstor.org/stable/3496837
Page Count: 5
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Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
Strain Identification of Spodoptera frugiperda (Lepidoptera: Noctuidae) Insects and Cell Line: PCR-RFLP of Cytochrome Oxidase C Subunit I Gene
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Abstract

A simple method was developed to analyze the two morphologically indistinguishable host-associated strains of the fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith). Total DNA extracted from the FAW corn and rice strains, as well as from the S. frugiperda cell line (SF9) was used to PCR amplify a 569 base pairs region of the mitochondrial gene cytochrome oxidase subunit I (COI). The amplified DNA from the Spodoptera corn strain and the SF9 cell line contained a restriction fragment length polymorphism (RFLP) marker, the MspI recognition site that was not present in the rice strain. This PCR-RFLP method does not require purification of mitochondrial DNA (mtDNA) or the use of radioactive isotopes, and differs from previous methods in that only a few nanograms of total DNA are needed to yield clear and accurate strain identification of individual insects. /// Un método simple ha sido desarrollado para analizar las dos cepas biológias, hospedero-dependiente, morfológicamente indistinguibles de Spodoptera frugiperda (J. E. Smith). El ADN total, extraído de insectos de S. frugiperda de las cepas de arroz y maíz, asi como también del cultivo de células SF9, fue utilizado para la amplificación, mediante el método de PCR. La región amplificada, de 569 pares de bases de la subunidad I del gen Citocromo Oxidase (COI), está localizada en el ADN mitocondrial. El ADN amplificado a partir de larvas y adultos de S. frugiperda de la cepa de maíz, contiene como marcador molecular, la secuencia reconocida por la enzima de restricción MspI, que no está presente en la cepa de arroz. Este método que combina la amplificación de ADN por PCR y subsecuente digestión enzimática (Polimorfismo de Longitud de Fragmento de Restricción-RFLP), no requiere la purifiación del ADN mitocondrial ni el uso de isótopos radioactivos. Se diferencia de otros métodos ya que sólo pocos nanogramos de ADN total son necesarios para obtener una identificación clara y precisa de insectos individuales.

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