DNA barcoding of five common stored-product pest species of genusCryptolestes(Coleoptera: Laemophloeidae)

Abstract

Several species of the genus Cryptolestes Ganglbauer, 1899 (Coleoptera: Laemophloeidae) are commonly found in stored products. In this study, five species of Cryptolestes, with almost worldwide distribution, were obtained from laboratories in China, Czech Republic and the USA: Cryptolestes ferrugineus (Stephens, 1831), Cryptolestes pusillus (Schönherr, 1817), Cryptolestes turcicus (Grouvelle, 1876), Cryptolestes pusilloides (Steel & Howe, 1952) and Cryptolestes capensis (Waltl, 1834). Molecular identification based on a 658 bp fragment from the mitochondrial DNA cytochrome c oxidase subunit I (COI) was adopted to overcome some problems of morphological identification of Cryptolestes species. The utility of COI sequences as DNA barcodes in discriminating the five Cryptolestes species was evaluated on adults and larvae by analysing Kimura 2-parameter distances, phylogenetic tree and haplotype networks. The results showed that molecular approaches based on DNA barcodes were able to accurately identify these species. This is the first study using DNA barcoding to identify Cryptolestes species and the gathered DNA sequences will complement the biological barcode database.

DNA barcoding, species-specific PCR and real-time PCR techniques for the identification of six Tribolium pests of stored products

“…A pair of universal forward LCO1490 (5′-GGTCAACAAA TCATAAAGATATTGG-3′) and reverse HCO2198 (5′-TAAACTTCAGGGTGACCA AAAAATCA-3′) primers were used for COI amplification15. PCR was performed based on methods by Wang et al 11. and was modified for half volume reactions containing 12.5 μl MasterMix with loading dye, 10 μl sterilized distilled water, 1.5 μl extracted DNA (approximately 20 ng μL −1 ), and 0.5 μl forward and reverse primers (10 μM).…”

COI barcode based species-specific primers for identification of five species of stored-product pests from genus Cryptolestes (Coleoptera: Laemophloeidae)

“…Use of five primer pairs for the conventional PCR system is described and their specificity and sensitivity in the identification process is evaluated. In comparison with the study of Wang et al (2014), the present identification system does not require sequencing, but only routine laboratory techniques such as DNA extraction, PCR and electrophoresis. Utilization of species-specific primers for identification is thus cost effective, time saving and does not compromise the reliability of identification.…”

“…Universal forward LCO1490 and reverse HCO2198 primers were used for COI amplification (Folmer et al, 1994). PCR was carried out based on methods by Wang et al (2014) in Veriti TM 96-well Thermal Cycler (ABI, USA). PCR products were confirmed by 1% agarose gel electrophoresis, stained with ethidium bromide (EB) and visualized under UV light (Gel Logic 212 PRO, Carestream Health, Inc.).…”

“…A short fragment of mitochondrial cytochrome c oxidase subunit I (COI) gene is widely used as the barcode sequence for animals and is frequently employed in applied entomology too (Armstrong & Ball, 2005;Nelson et al, 2007;Wells & Williams, 2007;deWaard et al, 2010;Jinbo et al, 2011;Morag et al, 2012;Wang et al, 2012;Jiang et al, 2013). Although species identification based on COI barcoding and species-specific primers has already been successfully applied for identification of different storage pests (Obrepalska-Steplowska et al, 2008;Nowaczyk et al, 2009;Yang et al, 2012Yang et al, , 2013Wang et al, 2014), more research needs to be conducted to enable this approach to be easily adopted for molecular identification of Cryptolestes species. Wang et al (2014) evaluated the accuracy of DNA barcoding based on COI gene for discrimination of the five most common storage Cryptolestes species.…”

“…Although species identification based on COI barcoding and species-specific primers has already been successfully applied for identification of different storage pests (Obrepalska-Steplowska et al, 2008;Nowaczyk et al, 2009;Yang et al, 2012Yang et al, , 2013Wang et al, 2014), more research needs to be conducted to enable this approach to be easily adopted for molecular identification of Cryptolestes species. Wang et al (2014) evaluated the accuracy of DNA barcoding based on COI gene for discrimination of the five most common storage Cryptolestes species. In that study, they clearly demonstrated that COI barcoding is a suitable alternative to traditional morphological identification of Cryptolestes species.…”