Isolation and characterization of microsatellites in the endoparasitic ichneumonid wasp carrying a polydnavirus Hyposote
- PDF / 162,767 Bytes
- 3 Pages / 595.276 x 790.866 pts Page_size
- 27 Downloads / 188 Views
TECHNICAL NOTE
Isolation and characterization of microsatellites in the endoparasitic ichneumonid wasp carrying a polydnavirus Hyposoter didymator Philippe Audiot • Ve´ronique Jouan • Marie Frayssinet • Anne-Nathalie Volkoff Denis Bourguet
•
Received: 23 July 2013 / Accepted: 17 September 2013 / Published online: 5 October 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract The wasp Hyposoter didymator (Hymenoptera, Ichneumonidae) parasitizes several agricultural pest moths and could therefore be used in biological control. The 454 FLX Titanium pyrosequencing technology was used to define two distinct sets of multiplex combining 14 polymorphic microsatellite loci: 10 (referred to as HD) located within the genome of H. didymator and 4 (referred to as HdIV) located within the Ichnovirus genome which is integrated into the wasp genome. Genotyping of two populations collected in France on Helicoverpa armigera revealed that most of the loci are independent and at Hardy–Weinberg equilibrium. Keywords Hyposoter didymator Helicoverpa armigera Pyrosequencing Parasitoid
Hyposoter didymator is a larval endoparasitoid wasp of many Lepidoptera larvae that carries a symbiotic polydnavirus (named H. didymator Ichnovirus, HdIV) integrated into its genome, insuring, like many other wasps, its vertical transmission. The most common host of H. didymator is the moth Helicoverpa armigera (Lepidoptera: Noctuidae), but this wasp can also parasitize other noctuid pest species. H. didymator may partly regulate their
P. Audiot (&) D. Bourguet CBGP, UMR INRA-IRD-CIRAD-Montpellier SupAgro, Campus International de Baillarguet, CS 30016, 34988 Montferrier-sur-Lez Cedex, France e-mail: [email protected] V. Jouan M. Frayssinet A.-N. Volkoff DGIMI, INRA-Universite´ Montpellier 2, Place Euge`ne Bataillon, CC101, 34095 Montpellier Cedex, France
populations and therefore can be used in biological control of those pests. Microsatellite markers of this parasitoid were isolated using the new generation 454 FLX titanium pyrosequencing technology (Malausa et al. 2011). Enrichment of microsatellite loci was carried out at Genoscreen (Lille, France) using the procedure described by Clamens et al. and Dumas et al. (in Arias et al. 2012). The selection of 454 FLX Titanium sequences for primer design was done using QDD software (Megle´cz et al. 2010). A total of 1,290 microsatellites loci were identified, amongst which 298 allowed designs of PCR amplification primers. Based on the expected sizes of amplification products, first attempts for polymorphism, multiplexing and quality of the chromatograms (details not shown), we chose 14 polymorphic microsatellite loci: 10 HD (referred to as HD loci) were located within the genome of H. didymator and 4 (referred to as HdIV) were located within the HdIV genome (Table 1). Those 14 loci were then amplified in two multiplex (M1 and M2) PCRs. PCRs were conducted in 10 lL reaction volume containing the Qiagen Multiplex PCR Master Mix (19)—with a final concentration of 3 mM of MgCl2 and an
Data Loading...
