Lilium spp. with symptoms of severe fasciation were observed in Southern and central Bohemia during the period 1999 – 2003. Nucleic acids extracted from symptomatic and asymptomatic plants were used in nested-PCR assays with primers amplifying 16S-23S rRNA sequences specific for phytoplasmas. The subsequent nested-PCR with phytoplasma-group specific primers followed by RFLP analyses and the 16S ribosomal gene sequencing, allowed to classify the detected phytoplasmas in the aster yellows group, subgroups 16SrI-B and 16SrI-C alone, and in mixed infection. Samples infected by 16SrI-C phytoplasmas showed different overlapping RFLP profiles after TruI digestion of R16F2/R2 amplicons. Two of these amplicons were sequenced, one of them directly and the other after cloning: sequence analyses and blast alignment confirmed the presence of two different overlapping patterns in samples studied. The sequences obtained were closely related respectively to operon A and operon B ribosomal sequences of the clover phyllody phytoplasma. Direct PCR followed by RFLP analyses of tuf gene with two restriction enzymes showed no differences from reference strain of subgroup 16SrI-C. Infection with aster yellows phytoplasmas of 16SrI-B subgroup in asymptomatic lilies cv. Sunray was also detected.
Molecular characterization of phytoplasmas in lilies with fasciation in the Czech Republic / Bertaccini A.; J. Fránová; S. Botti; D. Tabanelli. - In: FEMS MICROBIOLOGY LETTERS. - ISSN 0378-1097. - STAMPA. - 249:(2005), pp. 79-85. [10.1016/j.femsle.2005.06.001]
Molecular characterization of phytoplasmas in lilies with fasciation in the Czech Republic
BERTACCINI, ASSUNTA;BOTTI, SIMONA;
2005
Abstract
Lilium spp. with symptoms of severe fasciation were observed in Southern and central Bohemia during the period 1999 – 2003. Nucleic acids extracted from symptomatic and asymptomatic plants were used in nested-PCR assays with primers amplifying 16S-23S rRNA sequences specific for phytoplasmas. The subsequent nested-PCR with phytoplasma-group specific primers followed by RFLP analyses and the 16S ribosomal gene sequencing, allowed to classify the detected phytoplasmas in the aster yellows group, subgroups 16SrI-B and 16SrI-C alone, and in mixed infection. Samples infected by 16SrI-C phytoplasmas showed different overlapping RFLP profiles after TruI digestion of R16F2/R2 amplicons. Two of these amplicons were sequenced, one of them directly and the other after cloning: sequence analyses and blast alignment confirmed the presence of two different overlapping patterns in samples studied. The sequences obtained were closely related respectively to operon A and operon B ribosomal sequences of the clover phyllody phytoplasma. Direct PCR followed by RFLP analyses of tuf gene with two restriction enzymes showed no differences from reference strain of subgroup 16SrI-C. Infection with aster yellows phytoplasmas of 16SrI-B subgroup in asymptomatic lilies cv. Sunray was also detected.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.