PCR-RFLP-based method for reliable discrimination of cryptic species within Mecinus janthinus species complex (Mecinini, Curculionidae) introduced in North America for biological control of invasive toadflaxes

• Published in: BioControl (Dordrecht, Netherlands) Vol. 58; no. 4; pp. 563 - 573
• Main Authors: Toševski, Ivo, Jović, Jelena, Krstić, Oliver, Gassmann, Andre
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer-Verlag 01-08-2013; Springer Netherlands; Springer Nature B.V
• Subjects: Agriculture; Animal Biochemistry; Animal Ecology; Behavioral Sciences; Biological control; biological control agents; Biomedical and Life Sciences; COII gene; cost effectiveness; Cryptic species; Curculionidae; cytochrome-c oxidase; digestion; Entomology; Gene polymorphism; genes; Haplotypes; Introduced species; Invasive species; Life Sciences; Linaria vulgaris; Mecinus; Mecinus janthinus; Morphology; Physical characteristics; Plant Pathology; Polymerase chain reaction; Polymorphism; Restriction fragment length polymorphism; Taxonomy; Toadflax; North America; Europe; Canada; United States > US; Italy; Serbia; Switzerland; Germany; Molecular identification of biocontrol agent; Curculionidae; PCR-RFLP diagnostic assay; Invasive toadflaxes
• ISSN: 1386-6141; 1573-8248
• DOI: 10.1007/s10526-013-9506-2

Several populations of the stem-mining weevil Mecinus janthinus Germar species complex (Mecinini, Curculionidae), identified based on morphological characteristics, have been introduced in North America for the biological control of invasive toadflaxes of European origin: Linaria vulgaris Miller and L. dalmatica (L.) Miller (Plantaginaceae). According to the mitochondrial cytochrome oxidase subunit II (COII) gene haplotype divergence of Mecinus janthinus species complex, a total of 20 M. janthinus s.s., 3 M. janthinus s.l. of the ‘speciosa’ genotype and 29 M. janthiniformis haplotypes have been recorded across their native range in central and southeastern Europe. A polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) diagnostic assay of COII gene using Hpy188III and MnlI enzyme-mix, was developed for fast and cost-effective discrimination of these morphologically very similar cryptic weevil species. It is shown that digestion generates unique 4-fragment restriction profile in M. janthinus s.s., 2-fragment profile in M. janthiniformis and 3-fragment profile in M. janthinus s.l. ‘speciosa’ group of haplotypes, allowing precise identification of each species or genotype. The proposed method represents a practical tool for fast and accurate identification of the target biocontrol agents and should prevent using inappropriate weevil species in redistribution programs for biological control of invasive toadflax species.