MOLEKULÁRNÍ DETERMINACE VAJÍČEK A NYMF VYBRANÝCH MER OVOCNÝCH SADŮ RODU CACOPSYLLA METODOU REAL-TIME PCR [MOLECULAR DETERMINATION OF EGGS AND LARVAL STAGES OF SELECTED FRUIT TREE ORCHARD CACOPSYLLA PESTS BY REAL-TIME PCR]

Abstract

Cacopsylla mali, C. melanoneura, C. picta, C. pruni, C. pyri, and C. pyrisuga belong among important pests of fruit trees. They harm trees by sucking and transmitting pathogenic bacteria of the ‘Candidatus Phytoplasma’, 16SrX group, which leads to declining and death of fruit trees. Protection against these pests depends on their early reliable identification and monitoring in fruit nurseries and orchards. In general, the determination of adult psyllids is based on morphological criteria, however, it is difficult or even impossible to perform determination using eggs and nymphs. In this study, a recently developed real-time PCR method validated for the determination of adult psyllids, was applied for the identification of other developmental stages. A total, 46 samples of eggs and nymph instars N1 to N5 were successfully determined, indicating the PCR identification system can be reliably used also for these types of specimens. Specific PCRs detected three species – C. melanoneura, C. pyri a C. pruni which were also confirmed by sequencing of the cytochrome c oxidase subunit I (COI) gene. Thus, this innovative molecular method can be used for a quick and routine determination of selected Cacopsylla species infecting fruit trees in all developmental stages.

Keywords

• cacopsylla
• vector
• phytoplasma
• pcr detection