caa a22 4500 477101267 CHVBK 20180405111542.0 cr unu---uuuuu 170330e19960201xx s 000 0 eng 10.1007/BF00206246 doi (NATIONALLICENCE)springer-10.1007/BF00206246 Transient marker-gene expression during zygotic in-vitro embryogenesis of Brassica juncea (Indian mustard) following particle bombardment [Elektronische Daten] [Benedikt Kost, Nathalie Leduc, Christof Sautter, Ingo Potrykus, Gunther Neuhaus] A method has been established that allows the transfer of genes into single cells of excised globular-stage zygotic Brassica juncea L. embryos. The fate of single, genetically marked cells was followed during in-vitro embryogenesis. A simple and defined embryo culture medium has been designed on which zygotic B. juncea embryos, excised at the globular or at later stages, develop normally into mature, fully grown embryos. The smallest embryos which can be efficiently cultured are 30 μm long (embryo proper without suspensor) and are comprised of less than 20 cells. The embryos grow on the surface of solid medium without embedding and are freely accessible to microprojectile bombardment. Shooting at globular, transition and early heart-shaped embryos using both a particle inflow gun and a micro-targeting particle accelerator resulted in transient expression of genes encoding visible markers. For both particle-acceleration devices the shooting conditions have been optimised based on transient β-glucuronidase (GUS) expression. Bombarding embryos under optimal conditions had no deleterious effects on in-vitro embryogenesis. Multicellular GUS-expressing sectors were obtained, showing that cells can survive receiving a particle and resume normal development. The examination of these sectors has provided new information about the cell division patterns characterising early B. juncea embryogenesis. To be able to follow the development of particular genetically marked sectors, we tried to identify reporter genes that, in contrast to the uidA gene (which encodes GUS), can be non-destructively assayed in embryonic cells. Preliminary data has shown that expression of the firefly luciferase gene (Luc) can be detected in bombarded embryos without affecting their viability. Springer-Verlag, 1996 Brassica nationallicence Embryogenesis nationallicence Firefly luciferase nationallicence Microprojectile bombardment nationallicence CaMV : cauliflower mosaic virus nationallicence EMB : embryo culture medium nationallicence LUC ( Luc ) : firefly luciferase (gene) nationallicence GUS : β-glucuronidase nationallicence PIG : particle inflow gun nationallicence Kost Benedikt Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut Leduc Nathalie Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut Sautter Christof Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut Potrykus Ingo Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut Neuhaus Gunther Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut Planta Springer-Verlag 198/2(1996-02-01), 211-220 0032-0935 198:2<211 1996 198 425 https://doi.org/10.1007/BF00206246 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00206246 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kost Benedikt Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut NATIONALLICENCE 700 1- Leduc Nathalie Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut NATIONALLICENCE 700 1- Sautter Christof Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut NATIONALLICENCE 700 1- Potrykus Ingo Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut NATIONALLICENCE 700 1- Neuhaus Gunther Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092, Zürich, Switzerland aut NATIONALLICENCE 773 0- Planta Springer-Verlag 198/2(1996-02-01), 211-220 0032-0935 198:2<211 1996 198 425 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer