Articles
DETECTION OF MYCOPLASMALIKE ORGANISMS (PHYTOPLASMAS) IN RUBUS BY NESTED POLYMERASE CHAIN REACTION (PCR).
Article number
385_18
Pages
126 – 131
Language
Abstract
In 1991 a disease showing severe stunting and witches’ broom symptoms appeared in an orchard of Rubus fruticosus cv Hulltornleaf in Central Italy.
Two percent of the plants that showed stunted vegetation eventually declined and died within one or two years.
DNA was extracted from these plants and from symptomatic and asymptomatic wild R. fruticosus plants following procedures already described.
PCR was used to amplify 16S rDNA sequences specific to phytoplasmas.
When the primers R16F2/R2 were used, no visible amplification products were observed.
Using dilutions of these products, nested PCR experiments were performed with several primer pairs derived from 16S rDNA sequences internal to those amplified from R16F2/R2 and specific for different phylogenetic groups of phytoplasmas.
A specific product was obtained from all the symptomatic rubus samples when primers R16(V)F1/R1 were employed, indicating the presence of phytoplasmas belonging to the 16S rRNA group V, the same group as those infecting elm.
RFLP analysis of the amplified sequences indicated that the phytoplasmas infecting cultivated rubus were slightly different from those infecting American elm.
Two percent of the plants that showed stunted vegetation eventually declined and died within one or two years.
DNA was extracted from these plants and from symptomatic and asymptomatic wild R. fruticosus plants following procedures already described.
PCR was used to amplify 16S rDNA sequences specific to phytoplasmas.
When the primers R16F2/R2 were used, no visible amplification products were observed.
Using dilutions of these products, nested PCR experiments were performed with several primer pairs derived from 16S rDNA sequences internal to those amplified from R16F2/R2 and specific for different phylogenetic groups of phytoplasmas.
A specific product was obtained from all the symptomatic rubus samples when primers R16(V)F1/R1 were employed, indicating the presence of phytoplasmas belonging to the 16S rRNA group V, the same group as those infecting elm.
RFLP analysis of the amplified sequences indicated that the phytoplasmas infecting cultivated rubus were slightly different from those infecting American elm.
Authors
A. Bertaccini, M. Vibio, F. Gennari, S. Guerrini, A. Benni, I.-M. Lee
Keywords
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