A diagnostic test using the polymerase chain reaction is described for the detection of phytoplasma DNA in grapevines collected from South Australia and Victoria. Grapevines with Australian grapevine yellows disease tested positively for a phytoplasma but those with ‘restricted spring growth syndrome’ (formerly called ‘grapevine decline’) tested negatively. Restriction fragment length polymorphism analyses were done to determine the relationships between phytoplasmas of the Australian grapevine yellows and of representatives from both the aster yellows group (which includes phytoplasmas of grapevine yellows from Italy) and the elm yellows group (which includes phytoplasmas of flavescence dorée). Results showed that Australian grapevine yellows is associated with a unique phytoplasma that is more closely related to the phytoplasmas of the aster yellows group than to those of the elm yellows group.
Winter oilseed rape grown in several areas in South Bohemia showed symptoms of stunting, leaf reddening and extensive malformation of floral parts. Phytoplasmas were consistently observed by using electron microscopy only in phloem tissue of symptomatic plants. DNA isolated from infected and healthy control plants was used in PCR experiments. Primer pairs R16F2/R2, P1/P7 and rpF2/R2, amplifying, respectively, 16S rDNA, 16S rDNA plus spacer region and the beginning of the 23S and ribosomal protein gene L22 specific for phytoplasmas, were used. According to RFLP and sequence analyses of PCR products, the phytoplasma from rape was classified in the aster yellows phytoplasma group, subgroup 16SrI-B. The PCR products from the Czech phytoplasma-infected rape also had RFLP profiles identical to those of phytoplasma strains from Italian Brassica. This first molecular characterization of phytoplasmas infecting rape compared with strains from Brassica does not, however, clearly indicate differences among isolates of the same 16SrI-B subgroup. Further studies on other chromosomal DNA portions could help the research on host specificity or on geographical distribution of these phytoplasmas.
A survey for the presence of aster yellows-related phytoplasmas in the different life stages of Scaphoideus titanus was carried out by means of PCR and nested-PCR assays. Using a phytoplasma 16Srl group-specific primer pair followed by RFLP analysis of amplified products, we were able to detect and identify phytoplasmas from eggs, newly hatched nymphs, fourth and fifth instar nymphs and adults reared on ten phytoplasma-free Vicia faba seedlings. Two of these broad beans became infected, whereas PCR failed to detect phytoplasmas in the same ten plants before leafhopper rearing. These results suggest the possibility of transovarial transmission of aster yellows-related phytoplasmas in S. titanus.
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