Molecular Identification of ten species of stored-product psocids through microarray method based on ITS2 rDNA

Abstract

Stored-product psocids (Psocoptera: Liposcelididae) are cosmopolitan storage pests that can damage stored products and cause serious economic loss. However, because of the body size (~1 mm) of eggs, nymphs, and adults, morphological identification of most stored-product psocids is difficult and hampers effective identification. In this study, 10 economically important stored-product Liposcelis spp. psocids (Liposcelis brunnea, L. entomophila, L. decolor, L. pearmani, L. rufa, L.mendax, L. bostrychophila, L. corrodens, L. paeta, and L. tricolor) were collected from 25 geographic locations in 3 countries (China, Czech Republic, and the United States). Ten species-specific probes for identifying these 10 psocid species were designed based on ITS2 sequences. The microarray method and reaction system were optimized. Specificity of each of the ten probes was tested, and all probes were found suitable for use in identification of the respective10 Liposcelis spp. psocids at 66 °C. This method was also used to identify an unknown psocid species collected in Taian, China. This work has contributed to the development of a molecular identification method for stored-product psocids, and can provide technical support not only to facilitate identification of intercepted samples in relation to plant quarantine, but also for use in insect pest monitoring.

An advanced approach for rapid visual identification of Liposcelis bostrychophila (Psocoptera: Liposcelididae) based on CRISPR/Cas12a combined with RPA

“…Liposcelis bostrychophila is an important storage pest, which can damage stored products and cause serious economic loss (Liu et al 2017). Because of the tiny size of booklice eggs, nymphs, and adults, morphological identification is difficult and time-consuming.…”

“…The current availability of nucleic acid-based identification methods permits sensitive and accurate discrimination among morphologically identical booklice. For L. bostrychophila , several molecular identification methods have already been suggested, including DNA barcoding focusing on COI gene (Yang et al 2013), 16S rDNA gene (Li et al 2011), and ITS2 rDNA gene (Wei et al 2011), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology (Qin et al 2007, 2008), and microarray method (Liu et al 2017). Arif et al (2015) developed multiplex endpoint PCR and multiplex TaqMan qPCR technology to distinguish L. bostrychophila , L. brunnea , L. pearmani , L. decolor , and L. obscura .…”

Rapid identification of the invasive fall armyworm Spodoptera frugiperda (Lepidoptera, Noctuidae) using species-specific primers in multiplex PCR

“…However, the application of species-specific primers based on ITS sequences in insect identification remains rare. A few studies of mosquitos, thrips, and stored-product psocids have been proposed using ITS species-specific primers 35,36,41,[43][44][45][46] . The results of the present study suggest that the ITS1 sequence variations of Spodoptera with high interspecific variations of > 16% and low intraspecific divergence of < 1.2% (Table 1) can be favourable markers with which to develop species-specific primers for FAW identification.…”

DROP: Molecular voucher database for identification of Drosophila parasitoids

“…In addition to species identification, reference sequences from DROP may be used to create species-specific primers for the accurate identification of parasitoids, design multiplex PCR assays that rapidly distinguish species in natural or agricultural ecosystems (Ye et al, 2017), and apply high-throughput molecular identification diagnostics (Fagan-Jeffries et al, 2018). Applications of such specific primers have been used in bacteria, fungi, oomycetes and insect pests (Liu et al, 2017;Tedersoo et al, 2019;Tsai et al, 2020).…”