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Expression of viral resistance in transformed petunia plants regenerated in vitro
Year:
2005
Source of publication :
Acta Horticulturae
Authors :
Gera, Abdullah
;
.
Volume :
683
Co-Authors:
Ziv, M., RH Smith Institute of Plant Science, Hebrew University of Jerusalem, PO Box 12, Rehovot, 76100, Israel
Gandelman, M., RH Smith Institute of Plant Science, Hebrew University of Jerusalem, PO Box 12, Rehovot, 76100, Israel
Gera, A., Department of Virology, ARO Volcani Center, Bet Dagan 50250, Israel
Facilitators :
From page:
243
To page:
247
(
Total pages:
5
)
Abstract:
Vegetatively propagated trailing petunia is sensitve to virus infection. Attempts were made to develop a transformation protocol to introduce genes for virus resistance. Leaf segments from in vitro plants of two Cascadias varieties were cultured in vitro. Plants were regenerated from meristematic cell clusters that developed on segments cultured on a selected medium containing BA and NAA. Transformation was carried out with Agrobacterium. tumefaciens strains EHA105 and LBA4404. EHA105 carrying the plasmids pME504, pGA492PVY or pGA492PVYm and LBA4404 carrying pCMV N/B23, were used as a vector system for transformation. The plasmid pME504 carried the β-Glucuronidase (GUS) as a reporter gene and pGA492PVY carried the last three cistron of PVY (Protease-Replicase-Coat protein). The pGA492PVYm plasmid contained the same genes as described above with one distinction, the gene encoded the replicase had a mutation. The presence of acetosyringone during the incubation stage of the leaf segments increased the transformation efficiency. A total of 55 transgenic plants belonging to 55 independent lines transformed with a defective CMV replicase gene were challenged with CMV. Using the ELISA assay, two of the lines were found to be resistant to the virus while all the rest were susceptible. In addition, a total of 22 transgenic plants belong to 22 independent lines transformed with the plasmid pGA492PVY were also challenged by PVY. Using ELISA assay, two lines were found to have a delay in symptom development. A total of 23 transgenic plants transformed with the plasmid pGA492PVYM were also challenged by PVY. Using ELISA assay, only one line was found to be resistant to the virus. Southern blot analysis of resistant and susceptible transgenic plants showed the presence of the insert. The implication of these results is discussed.
Note:
Related Files :
A. tumefaciens
Cascadias petunia
CMV
Coat protein
PVY
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Related Content
More details
DOI :
Article number:
Affiliations:
Database:
Scopus
Publication Type:
Conference paper
;
.
Language:
English
Editors' remarks:
ID:
29699
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:48
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Scientific Publication
Expression of viral resistance in transformed petunia plants regenerated in vitro
683
Ziv, M., RH Smith Institute of Plant Science, Hebrew University of Jerusalem, PO Box 12, Rehovot, 76100, Israel
Gandelman, M., RH Smith Institute of Plant Science, Hebrew University of Jerusalem, PO Box 12, Rehovot, 76100, Israel
Gera, A., Department of Virology, ARO Volcani Center, Bet Dagan 50250, Israel
Expression of viral resistance in transformed petunia plants regenerated in vitro
Vegetatively propagated trailing petunia is sensitve to virus infection. Attempts were made to develop a transformation protocol to introduce genes for virus resistance. Leaf segments from in vitro plants of two Cascadias varieties were cultured in vitro. Plants were regenerated from meristematic cell clusters that developed on segments cultured on a selected medium containing BA and NAA. Transformation was carried out with Agrobacterium. tumefaciens strains EHA105 and LBA4404. EHA105 carrying the plasmids pME504, pGA492PVY or pGA492PVYm and LBA4404 carrying pCMV N/B23, were used as a vector system for transformation. The plasmid pME504 carried the β-Glucuronidase (GUS) as a reporter gene and pGA492PVY carried the last three cistron of PVY (Protease-Replicase-Coat protein). The pGA492PVYm plasmid contained the same genes as described above with one distinction, the gene encoded the replicase had a mutation. The presence of acetosyringone during the incubation stage of the leaf segments increased the transformation efficiency. A total of 55 transgenic plants belonging to 55 independent lines transformed with a defective CMV replicase gene were challenged with CMV. Using the ELISA assay, two of the lines were found to be resistant to the virus while all the rest were susceptible. In addition, a total of 22 transgenic plants belong to 22 independent lines transformed with the plasmid pGA492PVY were also challenged by PVY. Using ELISA assay, two lines were found to have a delay in symptom development. A total of 23 transgenic plants transformed with the plasmid pGA492PVYM were also challenged by PVY. Using ELISA assay, only one line was found to be resistant to the virus. Southern blot analysis of resistant and susceptible transgenic plants showed the presence of the insert. The implication of these results is discussed.
Scientific Publication
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