Genetic species identification of ecologically important planthoppers (Prokelisia spp.) of coastal Spartina saltmarshes using High Resolution Melting Analysis (HRMA)

Abstract

Phloem-feeding planthoppers of the genus Prokelisia rank among the most abundant and ecologically important browsers of coastal saltmarsh grasses of eastern North America and the Caribbean. Along the Spartina marshes of the northern Gulf of Mexico, the sympatric species P. marginata and P. dolus are the most abundant, but are difficult to distinguish from each other based solely on morphology. This study seeks to design a molecular assay based on High Resolution Melting Analysis (HRMA) as a fast, cost-effective alternative to differentiate these species. A 450 base pairs (bp) segment of cytochrome c oxidase subunit I (COI) was amplified and sequenced for representative samples of both species, and a short amplicon (SA) HRMA was designed based on the presence of fixed nucleotide differences between species found along a 60 bp segment of COI. The unambiguous identification of individual specimens of P. marginata or P. dolus was possible due to easily discernable differences in the melting temperatures of the two species along this mini barcode. This assay may prove useful for future genetic studies involving these species by preventing the overestimation of genetic diversity via inclusion of conspecifics, and in ecological studies by improving data on the effects of individual species of Prokelisia.

High‐throughput identification of tuna (Thunnus spp.) larvae in the Gulf of Mexico using unlabelled‐probe high‐resolution melting analysis

“…Real‐time (RT) PCR and HRMA using TaqMan probes were used to identify trout (McGlauflin et al ., 2010), and tunas (Bojolly et al ., 2017; Chuang et al ., 2012) and mackerels (Velasco et al ., 2013). HRMA has been employed to identify other taxa (Espinoza & Alvarado Bremer, 2019; Randall et al ., 2015; Winder et al ., 2011) and has been used to characterize fish populations (Smith et al ., 2013; Smith et al ., 2015). In this study, the authors investigate the ability of short‐amplicon (SA) HRMA and unlabelled probe (UP) HRMA to distinguish among tuna species present in the North Atlantic with an emphasis on the Gulf of Mexico.…”

Comparative Phylogeography, Historical Demography, and Population Genetics of Three Common Coastal Fauna in Spartina Marshes of the Northwestern Gulf of Mexico

“…P. pugio were compared against the mitogenome of five representatives of their sympatric sister species, P. vulgaris (Accession # JQ042300, JN674358, KP178999, KT959473, KT959519). P. marginata were compared against five representatives, each a different haplotype, of their sympatric sister species, P. dolus, that were sequenced by the authors for a previous study [65]. Generation time was assumed to be one year for F. grandis [33], two months for P. pugio [45], and 1.5 months for P. marginata [53].…”