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Isolating intact phytoplasma particles from plants for sequencing applications and for testing host responses to phytoplasmas
Year:
2019
Source of publication :
Phytopathogenic Mollicutes
Authors :
Assoline, Nofar
;
.
Bahar, Ofir
;
.
Volume :
Co-Authors:

Santos-Garcia, D. - Department of Entomology, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel;
Naor, V. - Shamir Research Institute, Katzrin, Israel;

Facilitators :
From page:
125
To page:
126
(
Total pages:
2
)
Abstract:

‘Candidatus Phytoplasma’ species are obligate plant parasites limited to the phloem tissue. The difficulties to grow phytoplasma in culture seriously impedes the study of these prokaryotes. In this study, a protocol to extract intact phytoplasmas from infected periwinkle plants using filtration, differential and density-gradient centrifugations was optimized. Confocal and transmission electron microscopy was used to visualize intact phytoplasmas in the preparation. Quantitative-PCR and DNA-seq analyses revealed that phytoplasma DNA was significantly enriched following the extraction procedure. Challenging periwinkle plants with phytoplasma preparation induced the expression of one defence-related gene. This study provides an optimized procedure for phytoplasma DNA enrichment and for extracting intact phytoplasmas that can be used to examine host responses to them.

Note:
Related Files :
DNA enrichment
microscopy
Phytoplasma
Sequencing
Show More
Related Content
More details
DOI :
10.5958/2249-4677.2019.00063.X
Article number:
0
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
46090
Last updated date:
02/03/2022 17:27
Creation date:
10/02/2020 16:26
Scientific Publication
Isolating intact phytoplasma particles from plants for sequencing applications and for testing host responses to phytoplasmas

Santos-Garcia, D. - Department of Entomology, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel;
Naor, V. - Shamir Research Institute, Katzrin, Israel;

Isolating intact phytoplasma particles from plants for sequencing applications and for testing host responses to phytoplasmas

‘Candidatus Phytoplasma’ species are obligate plant parasites limited to the phloem tissue. The difficulties to grow phytoplasma in culture seriously impedes the study of these prokaryotes. In this study, a protocol to extract intact phytoplasmas from infected periwinkle plants using filtration, differential and density-gradient centrifugations was optimized. Confocal and transmission electron microscopy was used to visualize intact phytoplasmas in the preparation. Quantitative-PCR and DNA-seq analyses revealed that phytoplasma DNA was significantly enriched following the extraction procedure. Challenging periwinkle plants with phytoplasma preparation induced the expression of one defence-related gene. This study provides an optimized procedure for phytoplasma DNA enrichment and for extracting intact phytoplasmas that can be used to examine host responses to them.

Scientific Publication
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