Abstract
winged sharpshooter, Homalodisca coagulata (Say), is challenging to study, not only because the bacterium is fastidious, but also because the insect does not survive well in captivity, and it will not feed from an artificial diet (i.e., parafilm sachet). We developed a simple and efficient transmission cycle for the study of X. fastidiosa transmission by H. coagulata, allowing collection of sufficient transmission data in 1 week. Specific numbers of cells can be detected both in the plant tissue and within the insect vector by realtime PCR (7). Real-time PCR is more sensitive than traditional PCR and is a single step process, thus reducing the possibility of error and speeding up detection time. A real-time PCR protocol for detecting the citrus variegated chlorosis strain of X. fastidiosa has already been established as a superior technique compared to traditional PCR (8). Real-time PCR was performed in a Rotor-Gene™ 3000 (Corbett Research, Mortlake, NSW, Australia) using iQ™ Supermix (Bio-Rad Laboratories, Hercules, CA, USA) in 20-µL reactions with 525 nM of each X.
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