96-well extraction technique for efficiently extracting Xyllela fastidiosa DNA from GWSS.

• Author(s): Bextine, Blake;
• Abstract: Xylella fastidiosa (Xf) is a xylem-limited bacterium that can cause disease in a wide range of plant hosts, including the Pierces disease of grapevines and citrus variegated chlorosis (CVC). Diseases caused by this pathogen threaten the wine grape and citrus industry in Texas. The primary insect vector of X. fastidiosa in Texas is the glassy-winged sharpshooter (GWSS), Homolodisca vitripennis. GWSS is polyphagous; feeding on a wide variety of ornamental, agricultural and native plant species. Grapevines are susceptible to the strains of X. fastidiosa that occur in Texas; however, the strain that causes CVC has not yet arrived in Texas. As GWSS is a major vector in both systems, a protocol that can simplify the detection of this pathogen in H. vitripennis is needed for early detection of the pathogen. In this study, different protocols were used to extract DNA from the insect mouthparts and foregut. The method that was most efficient, inexpensive and that allowed to extract a large sample size was the 96-well extraction technique used in conjunction with a silica-based DNA binding matrix. This extraction technique makes it economical feasible to screen a large number of insect samples to determine the presence or absence of X. fastidiosa, followed by strain identification.
• Publication Date: Mar 2007
• Journal: Proceedings Of The 55Th Annual Meeting Of The Southwestern Branch Of The Entomological Society Of America