Addressing the red flags in cochineal identification: the use of molecular techniques to identify cochineal insects that are used as biological control agents for invasive alien cacti

Invasive Cactaceae cause considerable damage to ecosystem function and agricultural practices around the world. The most successful biological control agents used to combat this group of weeds belong to the genus Dactylopius (Hemiptera: Dactylopiidae), commonly known as ‘cochineal’. Effective control relies on selecting the correct species, or in some cases, the most effective intraspecific lineage, of cochineal for the target cactus species. Many of the Dactylopius species are so morphologically similar, and in the case of intraspecific lineages, identical, that numerous misidentifications have been made in the past. These errors have resulted in failed attempts at the biological control of some cactus species. This study aimed to generate a multi-locus genetic database to enable the accurate identification of dactylopiids. Genetic characterization was achieved through the nucleotide sequencing of three gene regions (12S rRNA, 18S rRNA, and COI) and two inter-simple sequence repeats (ISSR). Nucleotide sequences were very effective for species-level and D. tomentosus lineage-level identification, but could not distinguish between the two lineages within D. opuntiae commonly used for biological control of various Opuntia spp. Fragment analysis through the use of ISSRs successfully addressed this issue. This is the first time that a method has been developed that can distinguish between these two D. opuntiae lineages. Using the methods developed in this study, biological control practitioners can ensure that the most effective agent species and lineages are used for each cactus target weed, thus maximizing the level of control.