AustralieFrV1, Main, Exploration, bibRecord, 00C497

Oligodeoxynucleotides as probes for in situ hybridization with transmission electron microscopy to specifically localize phytoplasma in plant cells

Identifieur interne : 00C497 ( Main/Exploration ); précédent : 00C496; suivant : 00C498

Oligodeoxynucleotides as probes for in situ hybridization with transmission electron microscopy to specifically localize phytoplasma in plant cells

Auteurs : J. Lherminier [France] ; R. G. Bonfiglioli [Australie] ; X. Daire [France] ; R. H. Symons [Australie] ; E. Boudon-Padieu [France]

Source :

Molecular and Cellular Probes [ 0890-8508 ] ; 1999.

RBID : ISTEX:DEC79778BCBF8DF514AD2BAF95754B2968A1779F

Descripteurs français

Wicri :
- topic : étiquetage, Maladie végétale.

English descriptors

Abstract

Abstract: Phytoplasmas are plant-pathogenic mollicutes restricted to phloem. They belong to several groups in a unique phylogenetic clade. Non-related phytoplasmas may infect the same plant species, often with similar symptoms. Hence methods are needed to specifically localize phytoplasmas and to study their multiplication and movement in their hosts. Conditions for post-embeddingin situhybridization (ISH) with transmission electron microscopy using oligodeoxynucleotides as probes for labelling of phytoplasmas in plant tissues have been searched. Sections of acrylic resin-embedded tissues of phytoplasma-infected periwinkle were submitted to ISH using digoxigenin or biotin-labelled oligoprobes (22 mers). These probes were the complementary sequences of primers used in group-specific polymerase chain reaction (PCR) amplification of 16S rDNA of stolbur and elm yellows phytoplasma, respectively. Together with preliminary digestion with pepsin, differentin situdenaturation conditions and formamide concentrations were tested. The grids were incubated in the hybridization mixture at 37°C overnight. Detection of hybridized material was performed with gold immunocytochemistry. Specificity of labelling was checked with appropriate controls. Stringency conditions could be found to ensure specific hybridization with such short probes. A specific labelling was obtained for stolbur phytoplasma on groups of mature as well as senescent phytoplasma cells. The results show that oligonucleotides may be used as probes for phytoplasma identification in post-embedding ISH with electron microscopy.

Url:

https://api.istex.fr/document/DEC79778BCBF8DF514AD2BAF95754B2968A1779F/fulltext/pdf

DOI: 10.1006/mcpr.1998.0213

Affiliations:

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Le document en format XML

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<term>Cultivar differences</term>
<term>Denaturing treatment</term>
<term>Dijon cedex</term>
<term>Doree</term>
<term>Electron micrographs</term>
<term>Electron microscopy</term>
<term>Experimental conditions</term>
<term>Flavescence</term>
<term>Flavescence doree</term>
<term>Formamide concentrations</term>
<term>Gold beads</term>
<term>Gold labelling</term>
<term>Gold particles</term>
<term>Gold resin</term>
<term>Grapevine</term>
<term>Grapevine yellows</term>
<term>Grid</term>
<term>Host plant</term>
<term>Hybridization</term>
<term>Hybridization buffer</term>
<term>Hybridization mixture</term>
<term>Hybridization signal</term>
<term>Labelling</term>
<term>Light label</term>
<term>Molecular cytology</term>
<term>Nucleic acids</term>
<term>Oligodeoxynucleotides</term>
<term>Periwinkle</term>
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<term>Plant tissue</term>
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<term>Same plant species</term>
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<term>Sense probe</term>
<term>Sequence analysis</term>
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<term>Stolbur group</term>
<term>Stolbur phytoplasma</term>
<term>Stringency conditions</term>
<term>Successive treatments</term>
<term>Target sequences</term>
<term>Tissue sections</term>
<term>Transmission electron microscopy</term>
<term>Unrelated probe</term>
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<term>Denaturing treatment</term>
<term>Dijon cedex</term>
<term>Doree</term>
<term>Electron micrographs</term>
<term>Electron microscopy</term>
<term>Experimental conditions</term>
<term>Flavescence</term>
<term>Flavescence doree</term>
<term>Formamide concentrations</term>
<term>Gold beads</term>
<term>Gold labelling</term>
<term>Gold particles</term>
<term>Gold resin</term>
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<term>Grapevine yellows</term>
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<term>Host plant</term>
<term>Hybridization</term>
<term>Hybridization buffer</term>
<term>Hybridization mixture</term>
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<term>Room temperature</term>
<term>Same plant species</term>
<term>Secondary structure</term>
<term>Senescent forms</term>
<term>Sense probe</term>
<term>Sequence analysis</term>
<term>Several groups</term>
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<term>Stolbur group</term>
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<term>Successive treatments</term>
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<term>Transmission electron microscopy</term>
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<front><div type="abstract" xml:lang="en">Abstract: Phytoplasmas are plant-pathogenic mollicutes restricted to phloem. They belong to several groups in a unique phylogenetic clade. Non-related phytoplasmas may infect the same plant species, often with similar symptoms. Hence methods are needed to specifically localize phytoplasmas and to study their multiplication and movement in their hosts. Conditions for post-embeddingin situhybridization (ISH) with transmission electron microscopy using oligodeoxynucleotides as probes for labelling of phytoplasmas in plant tissues have been searched. Sections of acrylic resin-embedded tissues of phytoplasma-infected periwinkle were submitted to ISH using digoxigenin or biotin-labelled oligoprobes (22 mers). These probes were the complementary sequences of primers used in group-specific polymerase chain reaction (PCR) amplification of 16S rDNA of stolbur and elm yellows phytoplasma, respectively. Together with preliminary digestion with pepsin, differentin situdenaturation conditions and formamide concentrations were tested. The grids were incubated in the hybridization mixture at 37°C overnight. Detection of hybridized material was performed with gold immunocytochemistry. Specificity of labelling was checked with appropriate controls. Stringency conditions could be found to ensure specific hybridization with such short probes. A specific labelling was obtained for stolbur phytoplasma on groups of mature as well as senescent phytoplasma cells. The results show that oligonucleotides may be used as probes for phytoplasma identification in post-embedding ISH with electron microscopy.</div>
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Oligodeoxynucleotides as probes for in situ hybridization with transmission electron microscopy to specifically localize phytoplasma in plant cells

Oligodeoxynucleotides as probes for in situ hybridization with transmission electron microscopy to specifically localize phytoplasma in plant cells

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri