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Lepidoptera: Sesiidae) Using Cytochrome Oxidase I Tennessee State University Digital Scholarship @ Tennessee State University Agricultural and Environmental Sciences Department of Agricultural and Environmental Faculty Research Sciences 7-1-2012 Molecular Identification of Synanthedonini Members (Lepidoptera: Sesiidae) Using Cytochrome Oxidase I Jason A. Hansen University of Tennessee, Knoxville William E. Klingeman University of Tennessee, Knoxville J. Kevin Moulton University of Tennessee, Knoxville Jason B. Oliver Tennessee State University Mark T. Windham University of Tennessee, Knoxville See next page for additional authors Follow this and additional works at: https://digitalscholarship.tnstate.edu/agricultural-and-environmental- sciences-faculty Part of the Entomology Commons Recommended Citation J. A. Hansen, W. E. Klingeman, J. K. Moulton, J. B. Oliver, M. T. Windham, A. Zhang, R. N. Trigiano, Molecular Identification of Synanthedonini Members (Lepidoptera: Sesiidae) Using Cytochrome Oxidase I, Annals of the Entomological Society of America, Volume 105, Issue 4, 1 July 2012, Pages 520–528, https://doi.org/10.1603/AN11028 This Article is brought to you for free and open access by the Department of Agricultural and Environmental Sciences at Digital Scholarship @ Tennessee State University. It has been accepted for inclusion in Agricultural and Environmental Sciences Faculty Research by an authorized administrator of Digital Scholarship @ Tennessee State University. For more information, please contact [email protected]. Authors Jason A. Hansen, William E. Klingeman, J. Kevin Moulton, Jason B. Oliver, Mark T. Windham, Aijun Zhang, and Robert N. Trigiano This article is available at Digital Scholarship @ Tennessee State University: https://digitalscholarship.tnstate.edu/ agricultural-and-environmental-sciences-faculty/208 SYSTEMATICS Molecular Identification of Synanthedonini Members (Lepidoptera: Sesiidae) Using Cytochrome Oxidase I J. A. HANSEN,1,2 W. E. KLINGEMAN,3 J. K. MOULTON,1 J. B. OLIVER,4 M. T. WINDHAM,1 5 1 A. ZHANG, AND R. N. TRIGIANO Downloaded from https://academic.oup.com/aesa/article/105/4/520/22145 by Tennessee State University user on 26 July 2021 Ann. Entomol. Soc. Am. 105(4): 520Ð528 (2012); DOI: http://dx.doi.org/10.1603/AN11028 ABSTRACT Many North American sesiid moths within Synanthodonini have been studied exten- sively because their feeding activity can cause detrimental economic and esthetic impacts to many commercially important ornamental and native plant species. Recent discoveries of nonnative clear- wing moth pest introductions [e.g., Synanthedon myopaeformis (Borkh.)], reinforce the need for reliable and accurate molecular diagnostic tools that can be used by nontaxonomic experts, particularly when juvenile life stages are recovered from infested host-plant tissues. Cytochrome oxidase I (cox I) previously has been used to successfully identify species and resolve species complexes. In this study, the cox I phylogeny inferred from sequences generated from 21 species of sesiid moths classiÞed within Synanthedonini conÞrms the close evolutionary relationship between sesiid species. As other authors have suggested in previous works, we observed that Synanthedon rileyana H. Edwards appears atypical for the genus, as it paired with Carmenta bassiformis (Walker) one node removed from, but not sister to, a large well-supported Synanthedon-rich clade. Sannina uroceriformis Walker and Po- dosesia Mo¨schler were observed nested deeply within the aforementioned well-supported clade (posterior probability [PP] of clade ϭ 100) comprised of all Synanthedon species sampled, except S. rileyana. Placement of these two taxa conßicts with results from previous morphological studies. These placements were immune from repeated attempts to delete perceived nearby long branches within the data set. Despite these few conßicts and overall low statistical support for most interspeciÞc and higher relationships, our data suggest that all species examined possess unique genetic signatures that lend themselves to accurate identiÞcation of all life history stages of these clearwing pests. KEY WORDS Lepidoptera, clearwing moth, barcoding, DNA Þngerprinting, woodborer The tribe Synanthedonini (Lepidopterea: Sesiidae) effective methods of control (Solomon and Dix 1979). was established by Niculescu 1964 based on adult Immature stages (i.e., eggs, larvae, and pupae), which morphological characters. The taxonomic division was are often encountered after feeding injury on plants supported in a subsequent revision of Sesiidae by using Þrst is observed, present particular difÞculties for pro- larval characters (MacKay 1968). Later, Naumann fessionals charged with pest control and regulatory (1971) included Synanthedon and other closely-re- action because morphological characters required for lated genera in a tribe he called Aegeriini, now con- accurate species identiÞcation often do not exist for sidered a synonym of Synanthedonini (Bradley et al. these life-stages, or may only be apparent to a taxo- 1972). Synanthedonini can be separated from all other nomic expert. North American tribes in Sesiidae by using wing ve- Larvae of several species within the genus Synan- nation. With over 87 species described, Synanthedo- thedon feed on fruit trees as well as native and non- nini is considered to be the most species-rich sesiid native ornamental plants. For example, larval feeding tribe in North America (Eichlin and Duckworth by S. exitiosa (Say) and S. pictipes (Grote and Robin- 1988). The economic importance attributed to many son), results in root, trunk, and branch damage that of its members has made these moth species subjects consequently reduces fruit yield and can kill host of research that has elucidated life-history details and plants. Likewise, the dogwood borer, S. scitula (Har- ris), is an increasingly important pest management 1 Department of Entomology and Plant Pathology, The University challenge in apple orchards, particularly where size- of Tennessee, 2431 Joe Johnson Dr., Knoxville, TN 37996. controlling rootstocks are used that result in borer- 2 Corresponding author, e-mail: [email protected]. 3 Department of Plant Sciences, The University of Tennessee, 2431 susceptible burr knots on stems, trunks, and graft Joe Johnson Dr., Knoxville, TN 37996. unions (Leskey and Bergh 2005). In addition to apple 4 TSU Otis Floyd Nursery Crops Research Station, 472 Cadillac Ln., and dogwood trees, the dogwood borer has perhaps McMinnville, TN 37110. the widest host plant range within Synanthedon. Its 5 USDAÐARS Plant Sciences Institute, Invasive Insect Biocontrol and Behavior Laboratory, BARC-West, 10300 Baltimore Ave., Belts- geographic distribution extends across the eastern ville, MD 20705. United States and to disjunct populations in the west- 0013-8746/12/0520Ð0528$04.00/0 ᭧ 2012 Entomological Society of America July 2012 HANSEN ET AL.: BARCODING IDENTIFICATION OF SYNANTHEDONINI 521 ern states of Colorado and Washington (Meyer et al. Materials and Methods 1988, Bergh et al. 2009, E. H. LaGasa, personal com- Taxon Sampling. Several well-documented clear- munication). In addition to nonnative clearwing moth wing moth pests were speciÞcally targeted, including introductions to North America during the past 150 yr, S. exitiosa, S. pictipes, S. scitula, S. viburni Engelhardt, the recent introduction of S. myopaeformis (Borkh.) and S. fatifera Hodges. Species-appropriate phero- from Europe reinforces the need for accurate molec- mone lures and light trapping were used to attract ular diagnostic tools to aid with identiÞcation of clear- male moths of desired taxa (Table 1). Where possible, wing pests, regardless of life stage (Eichlin and Duck- modiÞed Multipher-1 traps (Bio-Controˆle Ste-Foy, worth 1988, Philip 2006). QC, Canada) were located within habitats where host- Downloaded from https://academic.oup.com/aesa/article/105/4/520/22145 by Tennessee State University user on 26 July 2021 Mitochondrial genes are helpful diagnostic tools plant resources were known to occur. Traps were frequently used to quickly and reliably identify pest mounted Ϸ1 m (3.3 ft.) high on stands sited in partial species (Hebert et al. 2003, Armstrong and Ball 2005). noontime shade. Approximately 200 ml of ethanol was Combining gene sequence data with traditional mor- placed in a reservoir that was attached to the trap phological models eliminates sole reliance for taxo- funnel part with hot glue. Collection periods were for nomic differentiation by using morphological charac- durations not to exceed 7 d between reservoir reÞlls. ters that are often obscured or lost entirely by rough Pheromone lures were replaced at 6Ð8-wk intervals. handling, storage, and transport of specimens. Careful All moths were held and shipped to the University of dissection of damaged adult clearwing specimens Tennessee in Knoxville in vials containing 95% non- caught and preserved within pheromone traps can denatured ethanol. In the lab, species were identiÞed yield sufÞcient genetic material to allow subsequent using the descriptive keys of Eichlin and Duckworth species identiÞcation. Indeed, unique species-speciÞc (1988), then stored at Ϫ20ЊC until DNA was extracted. nucleotide arrangements within the mitochondrial ge- Specimen-collection data and the number of species nome have been used in several cases where morpho- from each site are reported (Table 1). Pinned voucher logical characters are either hard to discern or un- specimens have been placed in the Entomology and available (Ball and Armstrong 2006, Nwilene
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