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- Title
Identificación Molecular de Especies de Spalangia<sup>1</sup> Mediante los Espaciadores Internos Transcritos (ITS1 e ITS2) del ADNr Molecular Identification of Species of Spalangia<sup>1</sup> Using the Transcribed Internal Spacers (ITS1 and ITS2) of the rDNA
- Authors
Ávila-Rodríguez, Verónica; Nava-Camberos, Urbano; Alvarado-Gómez, Omar G.; Czaja, Alexander; Romero-Méndez, Ulises; Estrada-Rodríguez, José Luis
- Abstract
La identidad y filogenia molecular de las especies de Spalangia se determinaron mediante los marcadores intergénicos ITS1 e ITS2 del ADNr. Las especies identificadas fueron: Spalangia nigroaenea Curtis, Spalangia cameroni Perkins, y Spalangia endius Walker. Los tamaños de los fragmentos amplificados de la región ITS1 fueron: S. nigroaenea 970 pb, S. cameroni 720 pb, y S. endius 720 pb. El análisis de la digestión con la enzima AluI, mostró variación en los tamaños de los fragmentos de la región amplificada ITS1; para S. nigroaenea fueron de 620 y 350 pb, para S. endius fueron 430 y 290 pb, mientras que para S. cameroni no se observaron sitios de corte. Los tamaños de los fragmentos de la región intergénica ITS2 fueron: S. nigroaenea 720 pb, S. cameroni 520 pb, y S. endius 580 pb. Los tamaños de los fragmentos de restricción con AluI del gen ITS2 fueron: S. nigroaenea 510 y 210 pb, S. endius 300 y 280 pb, y S. cameroni 330 y 190 pb. Con base en estos resultados, las especies de Spalangia pueden distinguirse mediante los tamaños de fragmentos de las regiones ITS1 e ITS2 digeridos por la enzima AluI. El análisis filogenético con base en caracteres nucleotídicos utilizando Máxima Verosimilitud muestra una estrecha relación de las poblaciones de las especies de Spalangia de México y las de E.U.A. reportadas en el GenBank del NCBI. Los resultados del presente estudio son de relevancia ya que muestran la factibilidad de identificar de manera rápida y precisa las especies de Spalangia, mediante el análisis de restricción con la enzima AluI de los genes ITS1 e ITS2, lo cual es útil en el diseño e implementación de programas de control biológico de moscas. Species of parasitoids of the genus Spalangia are natural enemies of house flies, which are of great importance because of their diversity, abundance, wide geographical distribution, and feasibility of mass rearing. The correct identification of native parasitoids of house flies in a given region is the first step to design and implement a program of biological control of house flies, because this allows defining which species must be mass reared for their release, as well as to assess their impact and efficiency. However, the morphological identification of parasitoids is complex and usually requires specialists in taxonomy. Currently, molecular techniques are a quick and accurate alternative to the identification and characterization of Spalangia species. Internal transcribed spacers ITS1 and ITS2 of the rDNA have been used to determine the identity of the species, genetic variability, and phylogenetic inference of populations of Spalangia species. Therefore, the objective of the present study was to identify species of parasitoids of houseflies of the genus Spalangia and infer phylogenetic relationships through internal transcribed spacers ITS1 and ITS2 of the rDNA. Specimens of parasitoids of house fly pupae were collected in dairy stables from the Comarca Lagunera (states of Durango and Coahuila, Mexico). The parasitoids were preliminarily identified through morphological characters corresponding to the genus Spalangia. Identity and molecular phylogeny of Spalangia species were determined by the intergenic markers ITS1 and ITS2 of the rDNA. The species identified morphologically and molecularly were: Spalangia nigroaenea Curtis, Spalangia cameroni Perkins, and Spalangia endius Walker. The amplified fragment lengths of the ITS1 region were: S. nigroaenea 970 pb, S. cameroni 720 pb, and S. endius 720 pb. The digest analysis of the enzyme AluI showed variation in the sizes of the fragments of the amplified ITS1 region; thus 620 and 350 bp for S. nigroaenea, 430 and 290 bp for S. endius, but no court sites for S. cameroni. The sizes of the fragments of the intergenic region ITS2 were: S. nigroaenea 720 bp, S. cameroni 520 bp, and S. endius 580 bp. The restriction fragment lengths of the ITS2 gene with AluI were: S. nigroaenea 510 and 210 bp, S. endius 300 and 280 bp, and S. cameroni 330 and 190 bp. Based on these results, the Spalangia species can be distinguished by the fragment lengths of the regions ITS1 and ITS2 digested by the enzyme AluI. Phylogenetic analysis based on nucleotide characters using the Maximum Likelihood method shows a close relationship of the populations of Spalangia species from Mexico and the U.S.A reported in the GenBank of NCBI. The results of this study are important because they show the feasibility of quickly and accurately identifying Spalangia species, through restriction analysis with the enzyme AluI of ITS1 and ITS2 genes, which is useful in design and implementation of biological control programs of flies.
- Subjects
MEXICO; DURANGO (Mexico : State); COAHUILA (Mexico : State); MOLECULAR phylogeny; HOUSEFLY; MAXIMUM likelihood statistics; SPECIES; ENZYME analysis; FLY control
- Publication
Southwestern Entomologist, 2018, Vol 43, Issue 1, p209
- ISSN
0147-1724
- Publication type
Article
- DOI
10.3958/059.043.0113