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Open AccessArticle

Two Distinct Genotypes of Spissistilus festinus (Say, 1830) (Hemiptera, Membracidae) in the United States Revealed by Phylogenetic and Morphological Analyses

1
Section of Plant Pathology and Plant-Microbe Biology, School of Integrative Plant Science, Cornell University, Cornell AgriTech at the New York State Agricultural Experiment Station, Geneva, NY 14456, USA
2
(Current) Department of Plant and Environmental Sciences, College of Agriculture, Forestry, and Life Sciences, Clemson University, Clemson, SC 29634, USA
3
(Current) Interdepartmental Program in Translational Biology and Molecular Medicine, Baylor College of Medicine, Houston, TX 77030, USA
4
(Current) Departments of Chemistry and Biological Sciences, Harpur College of Arts and Sciences, Binghamton University, State University of New York, Binghamton, NY 13902, USA
5
Department of Entomology, Cornell University, Ithaca, NY 14853, USA
*
Author to whom correspondence should be addressed.
Insects 2020, 11(2), 80; https://doi.org/10.3390/insects11020080
Received: 6 January 2020 / Revised: 19 January 2020 / Accepted: 21 January 2020 / Published: 23 January 2020
Spissistilus festinus (Say, 1830) (Hemiptera: Membracidae) is a frequent pest of leguminous crops in the Southern United States, and a vector of grapevine red blotch virus. There is currently no information on the genetic diversity of S. festinus. In this study, populations of S. festinus were collected in 2015–2017 from various crops and geographic locations in the United States, and fragments of the mitochondrial cytochrome C oxidase 1 (mt-COI) gene and the nuclear internal transcribed spacer 2 (ITS2) region were characterized by polymerase chain reaction and sequencing. Maximum-likelihood and Bayesian analyses of the mt-COI and ITS2 sequences yielded similar phylogenetic tree topologies, revealing two distinct genetic S. festinus lineages with all of the specimens from California comprising one phylogenetic clade, alongside a single GenBank entry from Arizona, and all specimens from the Southeastern United States comprising a statistically-supported distinct clade, regardless of host and year of collection. The mt-COI gene fragment showed up to 10.8% genetic distance between the two phylogenetic clades. These results suggest the existence of two genotypes within S. festinus in the United States. The only distinct morphological trait between the two genotypes was a less elevated pronotum in the representative specimens from California, compared to the representative specimens from the Southeastern United States. Since this phenotypic feature is inconspicuous, a diagnostic polymerase chain reaction targeting a variable region of the mt-COI fragment was developed to reliably distinguish between the specimens of the two genotypes of S. festinus and to facilitate their specific identification.
Keywords: Spissistilus festinus; genetic barcoding; ITS2; mt-COI Spissistilus festinus; genetic barcoding; ITS2; mt-COI
MDPI and ACS Style

Cieniewicz, E.; Poplaski, V.; Brunelli, M.; Dombroskie, J.; Fuchs, M. Two Distinct Genotypes of Spissistilus festinus (Say, 1830) (Hemiptera, Membracidae) in the United States Revealed by Phylogenetic and Morphological Analyses. Insects 2020, 11, 80.

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