Applied and Environmental Microbiology · 1996 · 626 citations · 27 references
Life SciencesMolecular BiologyPlant SciencesGenomicsPlant GeneticsPlant Molecular Biology16S Ribosomal Ribonucleic Acid Analysis16S Ribosomal Ribonucleic Acid PhylogenyMolecular Markers In Plants16S Ribosomal Ribonucleic Acid Sequence Analysis16S Rrna Gene Phylogeny16S-23s Internal Transcribed Spacer AnalysisPlant Pathogen IdentificationPolymerase Chain Reaction (Pcr) AmplificationPrimer ApplicationPrimer DesignSpecies-specific Primer Design
In order to develop a diagnostic tool to identify phytoplasmas and classify them according to their phylogenetic group, we took advantage of the sequence diversity of the 16S-23S intergenic spacer regions (SRs) of phytoplasmas. Ten PCR primers were developed from the SR sequences and were shown to amplify in a group-specific fashion. For some groups of phytoplasmas, such as elm yellows, ash yellows, and pear decline, the SR primer was paired with a specific primer from within the 16S rRNA gene. Each of these primer pairs was specific for a specific phytoplasma group, and they did not produce PCR products of the correct size from any other phytoplasma group. One primer was designed to anneal within the conserved tRNA(Ile) and, when paired with a universal primer, amplified all phytoplasmas tested. None of the primers produced PCR amplification products of the correct size from healthy plant DNA. These primers can serve as effective tools for identifying particular phytoplasmas in field samples.
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