ELISA and dot-blot detection of flavescence dorée-MLO in individual leafhopper vectors during latency and inoculative state

Identification and Characterization of Phytoplasmal Genes, Employing a Novel Method of Isolating Phytoplasmal Genomic DNA

“…In the last 15 years, some progress has been made towards enabling the enrichment of phytoplasmal fractions from extracts of infected plants and insects (4,20,21,42,47,48). In some cases, it became possible to produce phytoplasma-specific polyclonal and monoclonal antibodies, greatly facilitating diagnosis (6,9). However, as detailed below, only a few phytoplasmal genes have been characterized.…”

Risk of a spreading a dangerous new grapevine disease, Flavescence dorée (FD) phytoplasma, and the distribution of its leafhopper vector (Scaphoideus titanus Ball) in Hungary

“…A Flavescence dorée (FD) fitoplazma és a Scaphoideus titanus kabó-cafaj esetében ezt a különös helyzetet azzal magyarázzák (Bressan et al 2005), hogy a vektorrovar és a fitoplazma co-evolúciója csak nagyon rövid idôre nyúlik vissza, hiszen 2000, Santini és Lucchi 1998). A fitoplazmát a rovar nyálmirigyébôl szerológiai módszer-rel (ELISA) és molekuláris módszerrel (PCR) is sikerült kimutatni, valamint az átviteli kísérletek is az átvitel lehetôségét bizonyítják (Boudon-Padieu et al 1989, Carraro et al 1994, Mori et al 2002.…”

Generation and characterization of monoclonal antibodies to Flavescence dorée phytoplasma: Serological relationships and differences in electroblot immunoassay profiles of Flavescence dorée and elm yellows phytoplasmas

“…FD-infected leafhoppers were obtained by feeding young adults on FD-diseased V. faba for 3 weeks (feeding access period), then on healthy Zea mays L. Infected leafhoppers were used after 8 weeks following the beginning of feeding access, when the phytoplasma titer is high in their body (Boudon-Padieu et al, 1989). Other phytoplasmas were maintained in periwinkle.…”

“…were individually crushed with a glass rod in a final volume of500pl PBS (phosphate-bufferedsaline: 137 mM NaC1, 2.7 mM KC1, 8 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.4) (Boudon-Padieu et al, 1989). The supernatant of a 2,900 g x 5 min centrifugation was used for ELISA (100 #1 per well).…”

“…Antisera to FD-phytoplasma have been produced (Boudon-Padieu et al, 1989) but thorough cross absorption was needed to get rid of antibodies to host components. However, western blots with polyclonal antibodies raised to phytoplasma-enriched fractions of FD-infected broadbean have demonstrated the presence in FD-infected leafhoppers of two main and 7-10 minor components which were not detected in healthy leafhopper control (Boudon-Padieu et al, 1987;Seddas et al, 1993).…”