An efficient Agrobacterium tumefaciens - mediated genetic transformation method for Centaurium erythraea via secondary somatic embrygenesis
2022
Authors:
Ćuković, KatarinaBogdanović, Milica
Simonović, Ana
Todorović, Slađana
Contributors
Ranđelović, VladimirStojanović-Radić, Zorica
Nikolić, Danijela
Jenačković Gocić, Dragana
Document Type:
Conference object (Published version)
,
© 2022 by the Department of Biology and Ecology, Faculty of Sciences and Mathematics, University of Niš
Metadata
Show full item recordAbstract:
A genetic transformation method via secondary somatic embryogenesis is described for Centaurium erythraea. Cotyledonary somatic embryos (cse) induced on leaf explants were used for inoculation with A. tumefaciens strain GV3101 containing the binary vector pXK7S2D that provides kanamycin resistance. Inoculated embryos were cultured on MS medium enriched with 2,4-dichlorophenoxyacetic acid and N-(2-chloro-4-pyridyl)-N’-phenylurea that promote somatic embryogenesis, with addition of acetosyringone. Cse were transferred on induction medium supplemented with cefotaxime and 5 or 10 mg/L kanamycin for selection. Embryogenic tissue, induced on primary cse explants during the selection period in the dark, was transferred to hormone-free medium maintaining the same antibiotic combination. Morphologically normal secondary cse that survived kanamycin exposure were isolated and subcultured on hormone-free medium containing cefotaxime and IBA to enhance germination. Fully regenerated plants were analyzed by Phire PCR to determine the transgene presence. Transformation efficiency was higher on media with 10 mg/L kanamycin (17.64%) as compared to 6.67% efficiency obtained on 5 mg/L. To the best of our knowledge, this is the first report on centaury transformation via secondary somatic embryogenesis, which offers an alternative to leaf or root explant transformation and provides an additional tool for investigating in vitro developmental pathways in this plant species.
Keywords:
agrobacterium transformation; somatic embryogenesis; 2,4-D; CPPU; secondary embryogenesis; kanamycinFunding / projects:
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200007 (University of Belgrade, Institute for Biological Research 'Siniša Stanković') (RS-MESTD-inst-2020-200007)
In:
- Ranđelović V, Stojanović-Radić Z, Nikolić D, Jenačković Gocić D, editors. Abstracts: 14th Symposium on the flora of southeastern Serbia and neighboring regions; 2022 Jun 26-29; Kladovo, Serbia. Niš : Department of Biology and Ecology, Faculty of Sciences and Mathematics, University of Niš; 2022. p. 112.