| Cloning of the spoT gene of "Candidatus Phlomobacter fragariae" and development of a PCR-restriction fragment length polymorphism assay for detection of the bacterium in insects. | |
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MedLine Citation:
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PMID: 10919809 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Marginal chlorosis is a new disease of strawberry in which the uncultured phloem-restricted proteobacterium "Candidatus Phlomobacter fragariae" is involved. In order to identify the insect(s) vector(s) of this bacterium, homopteran insects have been captured. Because a PCR test based on the 16S rRNA gene (rDNA) applied to these insects was unable to discriminate between "P. fragariae" and other insect-associated proteobacteria, isolation of "P. fragariae" genes other than 16S rDNA was undertaken. Using comparative randomly amplified polymorphic DNAs, an amplicon was specifically amplified from "P. fragariae"-infected strawberry plants. It encodes part of a "P. fragariae" open reading frame sharing appreciable homology with the spoT gene from other proteobacteria. A spoT-based PCR test combined with restriction fragment length polymorphisms was developed and was able to distinguish "P. fragariae" from other insect bacteria. None of the many leafhoppers and psyllids captured during several years in and around infected strawberry fields was found to carry "P. fragariae." Interestingly however, the "P. fragariae" spoT sequence could be easily detected in whiteflies proliferating on "P. fragariae"-infected strawberry plants under confined greenhouse conditions but not on control whiteflies, indicating that these insects can become infected with the bacterium. |
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Authors:
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X Foissac; J L Danet; L Zreik; J Gandar; J G Nourrisseau; J M Bové; M Garnier |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Applied and environmental microbiology Volume: 66 ISSN: 0099-2240 ISO Abbreviation: Appl. Environ. Microbiol. Publication Date: 2000 Aug |
Date Detail:
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Created Date: 2001-01-04 Completed Date: 2001-01-04 Revised Date: 2013-04-17 |
Medline Journal Info:
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Nlm Unique ID: 7605801 Medline TA: Appl Environ Microbiol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 3474-80 Citation Subset: IM |
Affiliation:
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Laboratoire de Biologie Cellulaire et Moléculaire, Institut National de la Recherche Agronomique et Université Victor Ségalen Bordeaux 2, Institut de Biologie Végétale Moléculaire, France. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Base Sequence Cloning, Molecular Fruit / microbiology Gammaproteobacteria / genetics, isolation & purification* Hemiptera / microbiology* Plant Diseases / microbiology Polymerase Chain Reaction* Polymorphism, Restriction Fragment Length* Pyrophosphatases / genetics* Random Amplified Polymorphic DNA Technique Sequence Analysis, DNA |
| Chemical | |
Reg. No./Substance:
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EC 3.1.7.2/guanosine-3',5'-bis(diphosphate) 3'-pyrophosphatase; EC 3.6.1.-/Pyrophosphatases |
| Comments/Corrections | |
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