NEW SINGLE-COPY NUCLEAR GENES FOR USE IN SCALE INSECT SYSTEMATICS

Abstract

Despite the advent of next-generation sequencing, the polymerase chain reaction (PCR) and Sanger sequencing remain useful tools for molecular identification and systematics. To date, molecular systematics of scale insects has been constrained by the paucity of loci that researchers have been able to amplify with available PCR primers. Due to the rapid molecular evolution of scale insects, “universal” primers, and even primers developed for their sister taxon the Aphidoidea, typically fail. We used transcriptome data for two diaspidids, Acutaspis umbonifera (Newstead) and Chrysomphalus aonidum (Linnaeus), together with a published aphid genome, to design novel PCR primer sets for scale insects. Our primers amplify fragments of eight single-copy genes: ATP-dependent RNA helicase (DHX8), translation initiation factor 5 (IF5X1), DNA replication licensing factor (Mcm2), double-strand break repair protein (MRE11A), serine/threonine- protein phosphatase (PPP1CB), DNA-directed RNA polymerase II (RNApII), ribonucleoside-diphosphate reductase (RRM1), signal recognition particle receptor (SRPα), neuronal PAS domain-containing protein 4 (NPAS4), and cleft lip and palate transmembrane protein 1 (TP1). Here we report the results of tests of amplification success and phylogenetic utility of these primer sets across the Diaspididae and nine other families of Coccomorpha.