Development of a system for the formation of transgenic somatic embryos in the liquid medium in Medicago truncatula
- Authors: Artemiuk A.M.1, Tvorogova V.E.1, Lutova L.A.1
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Affiliations:
- Saint Petersburg State University
- Issue: Vol 21 (2023): Спецвыпуск
- Pages: 9-9
- Section: Genetically modified organism. The Нistory, Achivements, Social and Environmental Riscs
- URL: https://journals.eco-vector.com/ecolgenet/article/view/568297
- DOI: https://doi.org/10.17816/ecogen568297
- ID: 568297
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Abstract
Somatic embryogenesis is the formation of embryos from plant’s somatic cells. It is widely used in biotechnology for reproduction of plants, studying of regeneration process and it also represents a convenient way to obtain transgenic plants. Currently, a solid medium is usually used for the formation of transgenic somatic embryos, which has a number of disadvantages.
We are developing a system for cultivating explants in a liquid medium for the transformation and formation of somatic embryos for Medicago truncatula. Unlike a solid medium, it should allow using petioles as explants, simplify the renewal of the medium, replace disposable cultivation containers with reusable ones, and also reduce the time required for the formation of somatic embryos.
Currently, the optimal concentration of hygromycin as a selective agent in such a system was found. Interestingly, it appeared to be lower than the selective hygromycin concentration in a solid medium.
The addition of cefotaxime to the medium reduces the number of somatic embryos formed, but does not completely suppress their formation. Thus, cefotaxime can be used to eliminate agrobacteria during transformation using this cultivation system.
Embryos with GUS overexpression transformed with this method were successfully obtained.
Keywords
Full Text
Somatic embryogenesis is the formation of embryos from plant’s somatic cells. It is widely used in biotechnology for reproduction of plants, studying of regeneration process and it also represents a convenient way to obtain transgenic plants. Currently, a solid medium is usually used for the formation of transgenic somatic embryos, which has a number of disadvantages.
We are developing a system for cultivating explants in a liquid medium for the transformation and formation of somatic embryos for Medicago truncatula. Unlike a solid medium, it should allow using petioles as explants, simplify the renewal of the medium, replace disposable cultivation containers with reusable ones, and also reduce the time required for the formation of somatic embryos.
Currently, the optimal concentration of hygromycin as a selective agent in such a system was found. Interestingly, it appeared to be lower than the selective hygromycin concentration in a solid medium.
The addition of cefotaxime to the medium reduces the number of somatic embryos formed, but does not completely suppress their formation. Thus, cefotaxime can be used to eliminate agrobacteria during transformation using this cultivation system.
Embryos with GUS overexpression transformed with this method were successfully obtained.
About the authors
Anastasiya M. Artemiuk
Saint Petersburg State University
Author for correspondence.
Email: nastyaart2004@gmail.com
Russian Federation, Saint Petersburg
Varvara E. Tvorogova
Saint Petersburg State University
Email: v.tvorogova@spbu.ru
ORCID iD: 0000-0002-0549-1457
cand. sci. (biol.), senior researcher, department of genetics and biotechnology
Russian Federation, Saint PetersburgLudmila A. Lutova
Saint Petersburg State University
Email: la.lutova@gmail.com
ORCID iD: 0000-0001-6125-0757
cand. sci. (biol.), senior researcher, department of genetics and biotechnology
Russian Federation, Saint Petersburg