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Genetic transformation of ornithogalum via particle bombardment and generation of pectobacterium carotovorum-resistant plants
Year:
2014
Source of publication :
Plant Science
Authors :
Cohen, Avner
;
.
Ion, Aurel
;
.
Lipsky, Alexander
;
.
Yedidia, Iris
;
.
Volume :
228
Co-Authors:
Lipsky, A., Department of Ornamental Horticulture, ARO, The Volcani Center, Derech Hamacabim 20, P.O. Box 6, Bet Dagan, Israel
Cohen, A., Department of Ornamental Horticulture, ARO, The Volcani Center, Derech Hamacabim 20, P.O. Box 6, Bet Dagan, Israel
Ion, A., Department of Ornamental Horticulture, ARO, The Volcani Center, Derech Hamacabim 20, P.O. Box 6, Bet Dagan, Israel
Yedidia, I., Department of Ornamental Horticulture, ARO, The Volcani Center, Derech Hamacabim 20, P.O. Box 6, Bet Dagan, Israel
Facilitators :
From page:
150
To page:
158
(
Total pages:
9
)
Abstract:
Bacterial soft rot caused by Pectobacterium carotovorum subsp. carotovorum (Pcc) is one of the most devastating diseases of Ornithogalum species. No effective control measures are currently available to use against this pathogen; thus, introduction of resistant genes via genetic transformation into this crop is a promising approach. Tachyplesin I, an antimicrobial peptide, has been shown to effectively control numerous pathogenic bacteria, including Pcc. In this study, liquid-grown cell clusters of Ornithogalum dubium and Ornithogalum thyrsoides were bombarded with a pCAMBIA2301 vector containing a celI leader sequence fused to a gene encoding tachyplesin I, a neomycin phosphotransferase (nptII) gene that served as a selectable marker and a β-glucuronidase (GUS) gene that served as a reporter. Selection was carried out in the dark in liquid medium containing 80. mg/L kanamycin. Regeneration was executed in the light after 6-14 months depending on the cultivar. Hundreds of transgenic plantlets were produced and their identity was confirmed through GUS activity assays. PCR and RT-PCR were used to confirm the presence of the target, reporter and selection genes in the divergent lines of plantlets. The resistance of the O. dubium plants to Pcc was evaluated in vitro, following infection with a highly virulent isolate from calla lily. Although control plantlets were completely macerated within a week, 87 putative transgenic subclones displayed varying levels of disease resistance. During three growing seasons in the greenhouse, the transgenic O. dubium lines grew poorly, whereas the transgenic O. thyrsoides plants grew similarly to non-transgenic plants. © 2014 Elsevier Ireland Ltd.
Note:
Related Files :
cyclopeptide
disease resistance
Genetics
genetic transformation
Ornithogalum
Show More
Related Content
More details
DOI :
10.1016/j.plantsci.2014.02.002
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
29254
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:45
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Scientific Publication
Genetic transformation of ornithogalum via particle bombardment and generation of pectobacterium carotovorum-resistant plants
228
Lipsky, A., Department of Ornamental Horticulture, ARO, The Volcani Center, Derech Hamacabim 20, P.O. Box 6, Bet Dagan, Israel
Cohen, A., Department of Ornamental Horticulture, ARO, The Volcani Center, Derech Hamacabim 20, P.O. Box 6, Bet Dagan, Israel
Ion, A., Department of Ornamental Horticulture, ARO, The Volcani Center, Derech Hamacabim 20, P.O. Box 6, Bet Dagan, Israel
Yedidia, I., Department of Ornamental Horticulture, ARO, The Volcani Center, Derech Hamacabim 20, P.O. Box 6, Bet Dagan, Israel
Genetic transformation of ornithogalum via particle bombardment and generation of pectobacterium carotovorum-resistant plants
Bacterial soft rot caused by Pectobacterium carotovorum subsp. carotovorum (Pcc) is one of the most devastating diseases of Ornithogalum species. No effective control measures are currently available to use against this pathogen; thus, introduction of resistant genes via genetic transformation into this crop is a promising approach. Tachyplesin I, an antimicrobial peptide, has been shown to effectively control numerous pathogenic bacteria, including Pcc. In this study, liquid-grown cell clusters of Ornithogalum dubium and Ornithogalum thyrsoides were bombarded with a pCAMBIA2301 vector containing a celI leader sequence fused to a gene encoding tachyplesin I, a neomycin phosphotransferase (nptII) gene that served as a selectable marker and a β-glucuronidase (GUS) gene that served as a reporter. Selection was carried out in the dark in liquid medium containing 80. mg/L kanamycin. Regeneration was executed in the light after 6-14 months depending on the cultivar. Hundreds of transgenic plantlets were produced and their identity was confirmed through GUS activity assays. PCR and RT-PCR were used to confirm the presence of the target, reporter and selection genes in the divergent lines of plantlets. The resistance of the O. dubium plants to Pcc was evaluated in vitro, following infection with a highly virulent isolate from calla lily. Although control plantlets were completely macerated within a week, 87 putative transgenic subclones displayed varying levels of disease resistance. During three growing seasons in the greenhouse, the transgenic O. dubium lines grew poorly, whereas the transgenic O. thyrsoides plants grew similarly to non-transgenic plants. © 2014 Elsevier Ireland Ltd.
Scientific Publication
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