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Agrobacterium rhizogenes-mediated transformation of a dioecious plant model Silene latifolia
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SYSNO ASEP 0502573 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Agrobacterium rhizogenes-mediated transformation of a dioecious plant model Silene latifolia Author(s) Hudzieczek, Vojtěch (BFU-R) ORCID
Čegan, Radim (BFU-R) RID, ORCID
Čermak, T. (US)
Bačovská, Nela (BFU-R)
Machálková, Zuzana (BFU-R)
Doležal, Karel (UEB-Q) RID, ORCID
Plíhalová, Lucie (UEB-Q) ORCID, RID
Voytas, D.F. (US)
Hobza, Roman (BFU-R) RID, ORCID
Vyskot, Boris (BFU-R) RID, ORCIDNumber of authors 10 Source Title New Biotechnology. - : Elsevier - ISSN 1871-6784
Roč. 48, JAN 25 2019 (2019), s. 20-28Number of pages 9 s. Publication form Print - P Language eng - English Country NL - Netherlands Keywords y-chromosome ; genetic-transformation ; biological-activity Subject RIV CE - Biochemistry OECD category Biochemistry and molecular biology Subject RIV - cooperation Institute of Experimental Botany - Biochemistry R&D Projects GA17-00567S GA ČR - Czech Science Foundation (CSF) LO1204 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support BFU-R - RVO:68081707 ; UEB-Q - RVO:61389030 UT WOS 000450366200004 DOI 10.1016/j.nbt.2018.04.001 Annotation Silene latifolia serves as a model species to study dioecy, the evolution of sex chromosomes, dosage compensation and sex-determination systems in plants. Currently, no protocol for genetic transformation is available for this species, mainly because S. latifolia is considered recalcitrant to in vitro regeneration and infection with Agrobacterium tumefaciens. Using cytokinins and their synthetic derivatives, we markedly improved the efficiency of regeneration. Several agrobacterial strains were tested for their ability to deliver DNA into S. latifolia tissues leading to transient and stable expression of the GUS reporter. The use of Agrobacterium rhizogenes strains resulted in the highest transformation efficiency (up to 4.7% of stable transformants) in hairy root cultures. Phenotypic and genotypic analyses of the T1 generation suggested that the majority of transformation events contain a small number of independent T-DNA insertions and the transgenes are transmitted to the progeny in a Mendelian pattern of inheritance. In short, we report an efficient and reproducible protocol for leaf disc transformation and subsequent plant regeneration in S. latifolia, based on the unique combination of infection with A. rhizogenes and plant regeneration from hairy root cultures using synthetic cytokinins. A protocol for the transient transformation of S. latifolia protoplasts was also developed and applied to demonstrate the possibility of targeted mutagenesis of the sex linked gene SlAP3 by TALENs and CRISPR/Cas9. Workplace Institute of Biophysics Contact Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244 Year of Publishing 2019 Electronic address https://doi.org/10.1016/j.nbt.2018.04.001
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