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ARS Home » Southeast Area » Stoneville, Mississippi » Southern Insect Management Research » Research » Publications at this Location » Publication #151526

Title: MOLECULAR DIFFERENTIATION OF TWO STRAINS OF THE PARASITOID ANISOPTERMALUS CALANDRAE HYMENOPTERA: PTEROMALIDAE) USING SPECIFIC PCR PRIMERS

Author
item Zhu, Yu Cheng
item Dowdy, Alan
item Baker, James

Submitted to: Journal of Entomological Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/17/2003
Publication Date: 12/22/2003
Citation: Zhu, Y., Dowdy, A.K., Baker, J.E. Molecular differentiation of two strains of the parasitoid anisoptermalus calandrae hymenoptera: pteromalidae) using specific pcr primers. Journal of Entomological Science. 39(1): 1-10 (January 2004)

Interpretive Summary: A strain (Bam) of Anisoptermalus clandrae collected from farm storage near Bamberg, SC, has developed more than 2000-fold higher resistance to malathion compared to a laboratory (Sav) strain. Because of the interest in developing beneficial insects with insecticide resistance, we have initiated studies to characterize the biochemical and molecular mechanisms involved in malathion resistance in the resistant strain of A. calandrae. In this study, we have examined molecular differences between two populations of A. calandrae. By using a combination of random amplification of polymorphic DNA (RAPD), PCR-cloning, and sequencing techniques, we have developed specific DNA markers to differentiate Bam strain from Sav strain of A. calandrae.

Technical Abstract: Molecular differences were studied by using polymerase chain reaction (PCR) and DNA sequencing techniques on two strains (Sav and Bam) of the parasitoid Anisopteromalus calandrae (Howard) (Hymenoptera: Pteromalidae) which differ in their sensitivity to organophosphate insecticides. Two DNA markers that are specific for Bam strain were developed and PCR-generated DNA markers that are specific for Ban strain were developed and PCR-generated DNA fragments were cloned and sequenced. Two DNA fragments unique to Bam strain contain 365 and 584 nucleotides. A pair of specific primers was designed from each fragment. PCR-amplification of the DNAs from individual wasps generated fragments of the expected sizes only in Bam strain. Study on F1 and F2 hybrids produced from crossing and backcrossing between resistant and susceptible strains indicated that these DNA markers are located on mitochondria and inherited exclusively maternally. Probes developed from these fragments may be used in assessing genetic information of natural populations and used in studies on physiological or biochemical differences between the strains of this beneficial insect.