Detecting Cacopsylla pyricola (Hemiptera: Psyllidae) in predator guts using COI mitochondrial markers

Abstract

Cacopsylla pyricola (Förster) is one of the most important pests of pear in North America, where several native predators have been considered for integrated pest management (IPM) programmes. Two molecular markers of 271 and 188 bp were developed from C. pyricola cytochrome oxidase I (COI) fragments, in order to study the detection of this species in the gut of arthropod predators. Primer sensitivity and the detection period for pear psylla remains in the guts of Anthocoris tomentosus Pericart were determined. The sensitivity threshold was defined at 10-5 dilution of a C. pyricola fifth-instar nymph in all samples. Predator adults were evaluated immediately after ingestion of one to five C. pyricola nymphs (t = 0) and after 2, 4, 6, 8, 16, 24 and 32 h. Detection of the presence of C. pyricola DNA always lasted longer using the shorter fragment and was observed after 32 h of digestion using both markers. The primers amplifying the 188 bp fragment amplified all four psyllid species tested, whereas the primers designed to amplify the 271 bp fragment did so exclusively for C. pyricola and its close relative, Cacopsylla pyri (Linnaeus). Both primers failed to amplify DNA from representative species of the Coccinellidae, Chrysopidae, Hemerobiidae, Anthocoridae, Miridae, Salticidae, Aphididae, Tetranychidae and the Tortricidae, suggesting their suitability for general trophic studies.

Detecting predation and scavenging by DNA gut-content analysis: a case study using a soil insect predator-prey system

“…Fresh prey 169 4 a Carrion prey 1 121 5 a 3 194 44 b 6 5 9 4 9 b 9 9 3 6 9 c The high sensitivity of PCR-based methods has been repeatedly confirmed (Chen et al 2000;Symondson 2002;Agusti et al 2003b). Under field conditions, eggs of cockchafers may be exposed to a higher predation pressure than larvae.…”

“…Previous studies utilizing PCR emphasize that successful identification of prey DNA strongly depends on the length of the target sequence. Fragments shorter than 250-350 bp survive digestion for >24 h, while detection of longer DNA fragments is limited (Zaidi et al 1999;Agusti et al 1999;Chen et al 2000;Hoogendoorn and Heimpel 2001;Agusti et al 2003b). This implies that time since feeding may be determined by using primers that amplify sequences of different length.…”

“…To date, detection of prey DNA 32 h postfeeding has only been described by Agusti et al (2003b) for Cacopsylla pyricola (Fo¨rster) (Hemiptera: Psyllidae) as prey of Anthocoris tomentosus Pericart (Hemiptera: Anthocoridae). Hoogendorn and Heimpel (2001) showed that PCR-based analysis of predation is influenced by temperature.…”

A Molecular Approach to Identifying the Natural Prey of the African Creeping Water BugNaucoris, A Potential Reservoir ofMycobacterium ulcerans

“…One limitation of most antibody- and DNA-based applications is that some knowledge of the potential prey is required. Antibodies target epitopes that are specific to proteins from target prey species, and most DNA methods employ species- or taxon-specific primers in PCR tests to determine the presence/absence of target prey species or taxa (e.g., Agustí et al 1999, 2000, 2003; Read 2002; Cuthbertson et al 2003; Jarman et al 2004; de León et al 2006). While these methods can be powerful and have been verified for their accuracy using laboratory feeding experiments (Chen et al 2000; Foltan et al 2005; Harper et al 2005, 2006; Sheppard et al 2005; Harwood et al 2007; McMillan et al 2007), a major limitation arises when there is little or no prior knowledge of the prey in their natural habitat.…”

Gut-content analysis of predatory phytoseiid mites using fluorescent-labeled polymerase chain reaction: age of spider-mite eggs influences detection rates

“…Recently, gut content analysis using molecular markers have been developed and used for many predatory species (e.g., Agustí et al, 2003;Greenstone, 2003;Hoogendoorn and Heimpel, 2001;Itou et al, 2013;Kamikawa et al, 2016;Symondson, 2002;Wari et al, 2014;Zhang et al, 2007). Phytoseiid mites─especially specialist species─tend to consume spider-mite eggs (Blackwood et al, 2001;Furuichi et al, 2005), and the small size of this preferred prey item hampers detection in the gut.…”