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The use of green fluorescent protein (GFP) improves Agrobacterium-mediated transformation of 'Spadona' pear (Pyrus communis L.)
Year:
2006
Source of publication :
Plant Cell Reports
Authors :
גולובוביץ', שרה
;
.
חנניה, אורי
;
.
יבלוביץ', זאב
;
.
פליישמן, משה
;
.
פרל, אבי
;
.
שליזרמן, לודמילה
;
.
Volume :
25
Co-Authors:
Yancheva, S.D., Plant Biotechnology Laboratory, Agricultural University, 12 Mendeleev Street, 4000 Plovdiv, Bulgaria
Shlizerman, L.A., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Golubowicz, S., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Yabloviz, Z., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Perl, A., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Hanania, U., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Flaishman, M.A., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Facilitators :
From page:
183
To page:
189
(
Total pages:
7
)
Abstract:
An efficient and reproducible system for Agrobacterium-mediated transformation of the pear (Pyrus communis L.) cultivar Spadona was developed. Leaf explants of in vitro propagated plants were cocultivated with the disarmed Agrobacterium strain EHA105 harboring the plasmid pME504, carrying the uidA-intron and nptII genes. Under selective conditions, 5% of the plantlets regenerated and were positively stained for GUS. However, most of the GUS-positive plants re-callused and subsequently died, leaving only 0.3-0.8% of these plantlets to reach maturity. In order to identify transformed shoots at early stages of regeneration, we introduced the green fluorescent protein (GFP) into the pear cultivar Spadona using the plasmid PZP carrying the nuclear-targeted GFP and nptII genes. High expression levels of GFP were detected in transgenic cells as early as 7 days after transformation. GFP marked-callii and transformed plants were observed after 14 and 24 days, respectively. Fluorescence microscopy screening of transformed plant material, under the selection of kanamycin, increased the transformation frequency to 3.0-4.0%. We conclude that the introduction of GFP improves the selection of transformed plants of Spadona pear. © Springer-Verlag 2005.
Note:
Related Files :
Agrobacterium
bacteria
Fruits
Genetics
genetic transformation
Growth, Development and Aging
metabolism
proteins
Pyrus communis
עוד תגיות
תוכן קשור
More details
DOI :
10.1007/s00299-005-0025-1
Article number:
Affiliations:
Database:
סקופוס
Publication Type:
מאמר
;
.
Language:
אנגלית
Editors' remarks:
ID:
23856
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:03
Scientific Publication
The use of green fluorescent protein (GFP) improves Agrobacterium-mediated transformation of 'Spadona' pear (Pyrus communis L.)
25
Yancheva, S.D., Plant Biotechnology Laboratory, Agricultural University, 12 Mendeleev Street, 4000 Plovdiv, Bulgaria
Shlizerman, L.A., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Golubowicz, S., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Yabloviz, Z., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Perl, A., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Hanania, U., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
Flaishman, M.A., Department of Fruit Trees Sciences, Institute of Horticulture, Volcani Center, P.O. Box 6, Bet Dagan, 50250, Israel
The use of green fluorescent protein (GFP) improves Agrobacterium-mediated transformation of 'Spadona' pear (Pyrus communis L.)
An efficient and reproducible system for Agrobacterium-mediated transformation of the pear (Pyrus communis L.) cultivar Spadona was developed. Leaf explants of in vitro propagated plants were cocultivated with the disarmed Agrobacterium strain EHA105 harboring the plasmid pME504, carrying the uidA-intron and nptII genes. Under selective conditions, 5% of the plantlets regenerated and were positively stained for GUS. However, most of the GUS-positive plants re-callused and subsequently died, leaving only 0.3-0.8% of these plantlets to reach maturity. In order to identify transformed shoots at early stages of regeneration, we introduced the green fluorescent protein (GFP) into the pear cultivar Spadona using the plasmid PZP carrying the nuclear-targeted GFP and nptII genes. High expression levels of GFP were detected in transgenic cells as early as 7 days after transformation. GFP marked-callii and transformed plants were observed after 14 and 24 days, respectively. Fluorescence microscopy screening of transformed plant material, under the selection of kanamycin, increased the transformation frequency to 3.0-4.0%. We conclude that the introduction of GFP improves the selection of transformed plants of Spadona pear. © Springer-Verlag 2005.
Scientific Publication
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