DNA-based methods for the detection and the identification of phytoplasmas in insect vector extracts

DNA extraction and storage methods have been evaluated with laboratory-reared leafhoppers and/or field-collected leafhoppers and psyllids. Detection of four different phytopathogenic phytoplasmas, belonging to three taxonomic groups, has been achieved by several direct or nested polymerase chain rea...

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Bibliographic Details
Published in:Molecular biotechnology 2002-09, Vol.22 (1), p.9-18
Main Authors: BOSCO, Domenico, PALERMO, Simona, MASON, Giovanna, TEDESCHI, Rosemarie, MARZACHI, Cristina, BOCCARDO, Guido
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Biotechnology
Deoxyribonucleic acid
DNA
DNA, Bacterial - analysis
DNA, Bacterial - isolation & purification
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Hemiptera - microbiology
In vitro gene amplification. Pcr technique
Insect Vectors - microbiology
Insects
Methods. Procedures. Technologies
Phytoplasma
Plasma
Polymerase Chain Reaction - methods
Polymorphism, Restriction Fragment Length
Reference Values
Reproducibility of Results
Restriction Mapping - methods
Sensitivity and Specificity
Sequence Analysis, DNA - methods
Species Specificity
Studies
Tenericutes - classification
Tenericutes - genetics
Tenericutes - isolation & purification
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Summary:DNA extraction and storage methods have been evaluated with laboratory-reared leafhoppers and/or field-collected leafhoppers and psyllids. Detection of four different phytopathogenic phytoplasmas, belonging to three taxonomic groups, has been achieved by several direct or nested polymerase chain reaction (PCR) methods with such DNA extracts. Reactions differed in both the 16/23S ribosomal primer pairs used and the specific assay and cycling conditions. Merits and possible hindrances of the various primer pairs, in relation to insect DNA extracts, are discussed. However, identification of the phytoplasma(s) necessarily relied on comparison of the polymorphism in length of the amplified DNA fragments obtained by restriction with appropriate endonucleases. Endonuclease digestion is crucial for determining the identity (subgroup affiliation) of phytoplasmas of the same groups that can be carried by an individual vector.
ISSN:1073-6085
1559-0305
1073-6085
DOI:10.1385/MB:22:1:009