Our aim was to establish an efficient and reproducible system for producing
synthetic seeds from recurrent somatic embryogenesis in long-term cultures of
Gentiana lutea L. This species is a vulnerable medicinal plant, protected
both at the national and international levels, and is included in different
Red Lists and Books. In vitro culture, as an alternative to classical methods
of preservation, allows for the cyclic multiplication of plant material and
short-, medium- and long-term preservation of tissue collections.
Biotechnological approaches allow for maintenance of the plant material in a
confined space and protection against biotic and abiotic factors. Somatic
embryogenesis (SE) is the most efficient way to regenerate plants, ensuring
material for preservation and fundamental research. In our experiment,
recurrent somatic embryogenesis was developed in long-term cultures in the
presence of sugar alcohols (mannitol, sorbitol) and in the absence of growth
factors. This process proceeded at a high rate, with adventive somatic
embryos being generated in a continuous process, followed by maturation,
germination and development into plants. To follow the somatic embryogenesis
process, histological samples were made. We used these embryogenic cultures
for synthetic seed production and medium-term conservation. The viability of
somatic embryos after moderate osmotic stress treatment was tested using TTC.
Our methodology relied on the induction of somatic embryogenesis in the
presence of auxins in the first cycle of in vitro cultures, long-term high
embryogenic culture maintenance in the presence of sugar alcohols and
synthetic seed production.
Recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. as a source for synthetic seed production for medium-term preservation
