Importance of Natural Enemies for Stink Bug Control

The laboratory of James D. Harwood will work, in collaboration with PI Ruberson (Department of Entomology, University of Georgia), to design, optimize and characterize species-specific molecular markers for Nezara viridula and Aridelus rufotestaceus. Protocols will follow those already developed in the Harwood laboratory for molecular identification of feeding habits (e.g., Harwood et al. 2007, 2009; Jaramillo et al. 2010; Chapman et al. 2010). In summary, to facilitate the design of the abovementioned primers, a data matrix will be constructed that contains COI sequences from a variety of closely related taxa. For those individuals sequenced at the University of Kentucky, forward and reverse COI sequences from the same individual will be aligned using AlignIR (v. 2.0, LI-COR Biosciences Inc., Lincoln, Nebraska, USA). Additionally, BLASTN searches (Karlin & Altschul 1990, 1993) of GenBank and the Barcode of Life Database (Ratnasingham & Hebert 2007) will be performed to determine whether the sequences significantly matched those of the same or closely-related species. Multiple sequence alignments will be performed by eye with MacClade (v. 4.08; Maddison & Maddison 2005). With the aid of the software tools available at the Primer3 web site (Rozen & Skaletsky 1998), the multiple alignment will be used to design a pair of primers for each species that will amplify a relatively small (