Abstract
Psocids of the genus Liposcelis (Psocoptera: Liposcelididae) are stored product pests that are difficult to identify morphologically. A molecular method based on Restriction Fragment Length Polymorphism (RFLP) of the PCR-amplified 16S rDNA gene was developed for the rapid discrimination of four common species (L. bostrychophila, L. entomophila, L. decolor, and L. paeta). Different developmental stages and populations (P.R. China and Czech Republic) were tested. One DNA fragment of about 500 bp in length was amplified from genomic DNA and the fragment was then digested using the restriction endonuclease DraI. Identification of the relevant banding pattern allowed all the developmental stages and both sexes to be discriminated in the species tested. The banding patterns of L. entomophila from all populations were identical, while the relevant restriction digests and sequence analysis confirmed that the Chinese and Czech populations of L. bostrychophila, L. decolor, and L. paeta differed. In conclusion, PCR-RFLP with one pair of primers (16Sar and 16Sbr) and one restrictive endonuclease, DraI, proved a reliable method for rapidly discriminating the Liposcelis species tested.
Highlights
In the last decade, psocids have become economically important pests of stored commodities (Ding et al, 2002; Beckett & Morton, 2003; Kuþerová et al, 2003; Nayak et al, 2003; Stejskal et al, 2003; Nayak, 2006)
Different DNA and Polymerase Chain Reaction (PCR)-based molecular markers have been developed for insect diagnostic purposes, including Random Amplified Polymorphic DNA (RAPD), Restriction Fragment Length Polymorphisms (RFLP), satellite DNA, and Inter-Simple Sequence Repeats (ISSR) (Li, 2007)
The sequencing results showed that the mutation points in 16S rDNA were consistent with the banding patterns of the electrophoresis map, except for some small fragments that could not be detected by agarose gel electrophoresis
Summary
Psocids (booklice) have become economically important pests of stored commodities (Ding et al, 2002; Beckett & Morton, 2003; Kuþerová et al, 2003; Nayak et al, 2003; Stejskal et al, 2003; Nayak, 2006). More than 60 sequences have been submitted to international DNA sequence databases, such as the DNA Data Bank of Japan (DDBJ), European Molecular Biology Laboratory (EMBL), and GenBank. Using these Liposcelis DNA sequences, we have analyzed a rapid discrimination method for Liposcelis from China based primarily on PCR-RFLP (Qin et al, 2007). The above discrimination investigations are extended by extracting genetic DNA from single individuals and using PCR-RFLP to discriminate between four species of Liposcelis originating from the People’s Republic of China (P.R. China) and Czech Republic (CZ), L. bostrychophila, L. entomophila, L. decolor, and L. paeta
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