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Inoculation of plants with begomoviruses by particle bombardment without cloning: Using rolling circle amplification of total DNA from infected plants and whiteflies
Year:
2010
Source of publication :
Journal of Virological Methods
Authors :
Gaba, Victor
;
.
Guenoune (Gelbart), Dana
;
.
Lapidot, Moshe
;
.
Sufrin-Ringwald, Tali
;
.
Volume :
168
Co-Authors:
Guenoune-Gelbart, D., Department of Vegetable Research, Volcani Center, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Sufrin-Ringwald, T., Department of Vegetable Research, Volcani Center, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Capobianco, H., Department of Plant Pathology, University of Florida, Gainesville, FL 32611, United States
Gaba, V., Department of Plant Pathology, Volcani Center, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Polston, J.E., Department of Plant Pathology, University of Florida, Gainesville, FL 32611, United States
Lapidot, M., Department of Vegetable Research, Volcani Center, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Facilitators :
From page:
87
To page:
93
(
Total pages:
7
)
Abstract:
A new system for inoculation of plants with begomoviral DNA without cloning or the use insect vectors is described. Total DNA extracted from begomovirus-infected plants was amplified by rolling circle amplification (RCA) using the bacteriophage phi29 DNA polymerase, and inoculated to plants by particle bombardment. Infection rates of up to 100% were obtained using this technique. This technique successfully inoculated all the begomoviruses evaluated: five bipartite (Bean golden yellow mosaic virus, Cabbage leaf curl virus, Squash leaf curl virus, Tomato mottle virus, Watermelon chlorotic stunt virus) as well as one monopartite (Tomato yellow leaf curl virus). The success of the technique was not dependent upon plant species. Four species from three plant families [. Phaseolus vulgaris (bean), Solanum lycopersicum (tomato), Cucurbita pepo (squash), and Citrullus lanatus (watermelon)], could all be inoculated by this technique. The success of the method was not dependent upon either the type or the age of the source of virus. Infectious DNA was obtained successfully from fresh, freeze-dried or desiccated plant material, from squashes of plant leaves on FTA cards, as well as from the insect vector. Plant material collected and dried as long as 25 years ago yielded infectious DNA by this method. In summary, this method can be used to obtain infectious DNA of single-stranded circular DNA viruses that can be activated for purposes of completing Koch's postulates, for preservation of pure virus cultures, and for many other applications where infectious DNA is required. © 2010 Elsevier B.V.
Note:
Related Files :
Animals
Citrullus lanatus
Cucurbita
Cucurbita pepo
Genetics
Plants
Tomato yellow leaf curl virus
Show More
Related Content
More details
DOI :
10.1016/j.jviromet.2010.04.022
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
19320
Last updated date:
02/03/2022 17:27
Creation date:
16/04/2018 23:28
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Scientific Publication
Inoculation of plants with begomoviruses by particle bombardment without cloning: Using rolling circle amplification of total DNA from infected plants and whiteflies
168
Guenoune-Gelbart, D., Department of Vegetable Research, Volcani Center, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Sufrin-Ringwald, T., Department of Vegetable Research, Volcani Center, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Capobianco, H., Department of Plant Pathology, University of Florida, Gainesville, FL 32611, United States
Gaba, V., Department of Plant Pathology, Volcani Center, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Polston, J.E., Department of Plant Pathology, University of Florida, Gainesville, FL 32611, United States
Lapidot, M., Department of Vegetable Research, Volcani Center, Agricultural Research Organization, P.O. Box 6, Bet Dagan 50250, Israel
Inoculation of plants with begomoviruses by particle bombardment without cloning: Using rolling circle amplification of total DNA from infected plants and whiteflies
A new system for inoculation of plants with begomoviral DNA without cloning or the use insect vectors is described. Total DNA extracted from begomovirus-infected plants was amplified by rolling circle amplification (RCA) using the bacteriophage phi29 DNA polymerase, and inoculated to plants by particle bombardment. Infection rates of up to 100% were obtained using this technique. This technique successfully inoculated all the begomoviruses evaluated: five bipartite (Bean golden yellow mosaic virus, Cabbage leaf curl virus, Squash leaf curl virus, Tomato mottle virus, Watermelon chlorotic stunt virus) as well as one monopartite (Tomato yellow leaf curl virus). The success of the technique was not dependent upon plant species. Four species from three plant families [. Phaseolus vulgaris (bean), Solanum lycopersicum (tomato), Cucurbita pepo (squash), and Citrullus lanatus (watermelon)], could all be inoculated by this technique. The success of the method was not dependent upon either the type or the age of the source of virus. Infectious DNA was obtained successfully from fresh, freeze-dried or desiccated plant material, from squashes of plant leaves on FTA cards, as well as from the insect vector. Plant material collected and dried as long as 25 years ago yielded infectious DNA by this method. In summary, this method can be used to obtain infectious DNA of single-stranded circular DNA viruses that can be activated for purposes of completing Koch's postulates, for preservation of pure virus cultures, and for many other applications where infectious DNA is required. © 2010 Elsevier B.V.
Scientific Publication
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