Detection of aster yellows phytoplasma in false flax based on PCR and RFLP

False flax ( Camelina sativa L.) plants were found to be infected with a yellows-type disease caused by a phytoplasma in experimental plots at the Edmonton Research station, Alberta, Canada. Typical phytoplasmas were detected in the phloem cells in ultrathin sections from leaf midrib tissues examine...

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Veröffentlicht in:Microbiological research 2001, Vol.156 (2), p.179-184
Hauptverfasser: Khadhair, A.-H., Tewari, J.P., Howard, R.J., Paul, V.H.
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Howard, R.J.
Paul, V.H.
description False flax ( Camelina sativa L.) plants were found to be infected with a yellows-type disease caused by a phytoplasma in experimental plots at the Edmonton Research station, Alberta, Canada. Typical phytoplasmas were detected in the phloem cells in ultrathin sections from leaf midrib tissues examined by electron microscopy. These observations were supported by polymerase chain reaction (PCR) using two primer pairs, R16 F2n/R2 and R16(1)F1/R1, derived from phytoplasma rDNA sequences. Aster yellows (AY) and pota-to witches'-broom (PWB) phytoplasma DNA samples served as controls and were used to study group relatedness. In a direct PCR assay, DNA amplification with universal primer pair R16F2n/R2 gave the expected PCR products of 1.2 kb. Based on a nested-PCR assay using the latter PCR products as templates, and a specific primer pair, R16(1)F1/R1, designed on the basis of AY phytoplasma rDNA sequences, a PCR product of 1.1 kb was obtained from each phytoplasma-infected false flax and AY sample, but not from PWB phytoplasma and healthy controls. DNA amplification with specific primer pair R16(1)F1/R1 and restriction fragment length polymorphism indicated the presence of AY phytoplasma in the infected false flax sample. This is the first reported characterization of AY phytoplasma in false flax.
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source MEDLINE; Elsevier ScienceDirect Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals
subjects Acholeplasmataceae - genetics
Acholeplasmataceae - isolation & purification
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Biotechnology
Brassicaceae - microbiology
Camelina sativa
detection
DNA, Bacterial - analysis
DNA, Ribosomal - genetics
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
In vitro gene amplification. Pcr technique
Methods. Procedures. Technologies
Microbiology
Microscopy, Electron
PCR RFLP
phytoplasma
Plant Diseases - microbiology
Polymerase Chain Reaction - methods
Polymorphism, Restriction Fragment Length
RNA, Ribosomal, 16S - genetics
title Detection of aster yellows phytoplasma in false flax based on PCR and RFLP
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