title: Real-time PCR for specific detection of three phytoplasmas from the apple proliferation group.
abstract: In this chapter, we describe a real-time PCR detection system for fast, reliable, specific, and sensitive detection and discrimination of 'Candidatus Phytoplasma mali', 'Ca. P. prunorum', and 'Ca. P. pyri' from the 16SrX (apple proliferation-AP) group. These phytoplasmas are causal agents of fruit tree diseases within the Rosaceae family, namely apple proliferation, European stone fruit yellows, and pear decline. The assays use (hydrolysis) TaqMan(®) minor groove binder probes. The panel of assays comprises the same set of primers and specific probes for species-specific amplification, and an additional set of primers and probe for 18S rRNA as an endogenous quality control of DNA extraction. The assays described can be used in routine phytoplasma surveys and in certification programmes.
authors.affiliation: National Institute of Biology, Ljubljana, Slovenia. natasa.mehle@nib.si
authors.forename: Nataša
authors.initials: N
authors.lastname: Mehle
authors.forename: Petra
authors.initials: P
authors.lastname: Nikolić
authors.forename: Kristina
authors.initials: K
authors.lastname: Gruden
authors.forename: Maja
authors.initials: M
authors.lastname: Ravnikar
authors.forename: Marina
authors.initials: M
authors.lastname: Dermastia
mesh_terms: D019295:Computational Biology / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics; D005638:Fruit / Q000382:microbiology; D027845:Malus / Q000382:microbiology; D045582:Phytoplasma / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D010935:Plant Diseases / Q000382:microbiology; D012335:RNA, Ribosomal / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*
chemical_list: D004269:DNA, Bacterial; D012335:RNA, Ribosomal