Diagnostic analysis of phytoplasma in plants and insects using 454 pyrosequencing

Detection of phytoplasma infections in varying plant species and their insect host is difficult using standard molecular techniques. Lethal Yellowing is a disease of palm trees in which phytoplasma are the causative agents. For successful detection multiple extraction techniques were employed to obtain phytoplasma DNA from insects captured off infected palm trees. Concurrently, palm tissue was sampled for phytoplasma DNA using the same extraction techniques. Polymerase chain reaction was employed to amplify phytoplasma DNA with a known working primer set. Gel electrophoresis was performed to confirm positive results. Additionally, presence of bacterial composition within the insects of study was completed using 454 Pyrosequencing. This provided supporting sequence data, evidence of phytoplasma presence. Through development of better diagnostics for phytoplasma disease more information can be gathered to prevent infections.