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Agrobacterium rhizogenes-mediated transformation of a dioecious plant model Silene latifolia

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, ORCID}
Number of authors	10
Source Title	New Biotechnology. - : Elsevier - ISSN 1871-6784 Roč. 48, JAN 25 2019 (2019), s. 20-28
Number 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

Number of the records: 1