National Institute of Biology, Vecna pot 111, SI-1000, Ljubljana, Slovenia
*Corresponding author e-mail: Nataša Mehle, natasa.mehle@nib.si
Online published on 24 July, 2014.
The first droplet-digital-PCR-based absolute quantification of “flavescence dorée” phytoplasma, agent of a quarantine phytoplasma yellows in grapevine was performed. Quantitative PCR that targets the secY gene was transferred to droplet digital PCR and used for absolute quantification of “flavescence dorée” phytoplasma, without the need for calibration curves. The sensitivity of the two assays compares well and was shown that it could be used for quantification and quality control of DNA based on in-house reference materials typically used in diagnostics and metrological laboratories. This new tool has great potential for monitoring phytoplasma kinetics, such as there relationship with the progress of an infection, and variations of the phytoplasma titer through the season and screening plants for resistance.
“Flavescence dorée”, phytoplasmas, droplet digital PCR, quantitative PCR, DNA quantification