Mission possible: an optimised protocol for the unbarcodable Ceraphronoidea (Hymenoptera)

Abstract

DNA barcodes provide a reliable and efficient solution to resolving cryptic species complexes and accelerate species discoveries. The superfamily Ceraphronoidea (Hymenoptera) is a group of parasitoid wasps for which a barcoding approach could be of great help, if it were not for the very poor results. The inability to obtain barcodes for the majority of treated ceraphronoids halts progress on the taxonomy of this hyperdiverse parasitoid group. We here present a working protocol for the barcoding of ceraphronoid wasps which yields a first-time over 90% success rate.

Afrotropical Ceraphronoidea (Insecta: Hymenoptera) put back on the map with the description of 88 new species

“…The material had not been adequately stored to facilitate DNA extraction and sequencing. Further, it has been difficult to retrieve DNA and, for example, DNA barcode data from ceraphronoids; a problem that might be solved or improved upon by a very recently published protocol that was not available to us (Vasilita et al 2022). Other techniques, such as target DNA enrichment or whole genome sequencing, allow for generating sequence data even from small, poor-quality DNA, however, for specimens as small as ceraphronoids they are expensive and still in their infancy.…”

Surprising morphological diversity in ceraphronid wasps revealed by a distinctive new species of Aphanogmus (Hymenoptera: Ceraphronoidea)

“…For DNA barcoding, the protocol developed by Vasilița et al (2022) was used. All sequences are deposited at Barcode of Life Data (BOLD) Systems (Ratnasingham & Hebert 2007) (DOI: https://doi.org/10.5883/DS-CERAPKR), as well as in GenBank, the individual IDs for which are given in the type material section in the results.…”

Characterization of the Mitogenome of the Genus Dendrocerus Ratzeburg (Hymenoptera: Megaspilidae) with the Specific Designed Primers

“…Although effective in species delimitation, species identification by male genitalia has limitations, such as failure in matching the two sexes of the same species. Combining molecular identification can address this issue, but there are challenges in obtaining DNA barcodes (COX1) in Megaspilidae using universal primers due to the small size of the parasitoid, resulting in reduced amounts of extracted DNA and mixed DNA originating from the parasitioid and its host [18]. Vasilita et al (2022) [18] developed an optimized barcoding protocol for Ceraphronoidea, but the success rate of barcoding dropped as more samples were tested, and its applicability in Megaspilidae is not clear.…”

“…Combining molecular identification can address this issue, but there are challenges in obtaining DNA barcodes (COX1) in Megaspilidae using universal primers due to the small size of the parasitoid, resulting in reduced amounts of extracted DNA and mixed DNA originating from the parasitioid and its host [18]. Vasilita et al (2022) [18] developed an optimized barcoding protocol for Ceraphronoidea, but the success rate of barcoding dropped as more samples were tested, and its applicability in Megaspilidae is not clear. This suggests that new specific barcode primers designed with reference to complete mitochondrial genomes may be required.…”