Description
PATHOGEN INFORMATION
Pear decline phytoplasma (Candidatus Phytoplasma pyri) is the causative agent of serious diseases in apple and pear trees, causing significant economic damage to fruit growing across Europe. These bacteria belong to the 16SrX taxonomic group and are considered quarantine organism for Eurasian Economic Union countries, and member countries of European and Mediterranean Plant Protection Organization (EPPO A2 list). Pyrus spp., Malus spp., Cydonia spp. are the most economically significant hosts for Candidatus Phytoplasma pyri.
ASSAY PRINCIPLE
«ALSENSE Phytoplasma pyri» Reagent kit has been developed based on recommendations of the European and Mediterranean Plant Protection Organization (EPPO). The reagent kit is a multiplex, real-time PCR-based assay. The target for amplification is the 16S ribosomal RNA gene region of phytoplasmas of the 16SrX group. The internal control (IC) is designed based on the 18S gene sequence of apple and pear ribosomal RNA. The TaqMan® probe to a region of the Candidatus Phytoplasma pyri genome is labeled with a FAM fluorophore dye on the 5′-end. The IC-specific probe is labelled with a HEX fluorophore, allowing the user to validate the DNA extraction step from plants.
SAMPLING RECOMMENDATIONS
«ALSENSE Phytoplasma pyri» is validated for the simultaneous detection of Candidatus Phytoplasma pyri and plant DNA by real-time PCR. It is recommended to use samples of shoots, petioles and midveins of leaves with disease symptoms for preliminary DNA extraction. Fresh plant tissue samples are optimal, but storage at minus 20 to minus 80°C is acceptable.