Recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. as a source for synthetic seed production for medium-term preservation

Abstract

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

Role of Biotechnology for Protection of Endangered Medicinal Plants

“…Somatic embryogenesis is another morphogenetic pathway for regeneration of plants, which is considered the most efficient way to regenerate plants [122]. In contrast to organogenesis when the buds and shoots are not formed obligatory from one cell, a somatic embryo derives from a single cell.…”

“…However, somatic embryogenesis is more difficult to obtain. Nevertheless, it was successfully induced in a number of Gentiana species: Gentiana lutea [122,123], Gentiana crassicaulis, Gentiana cruciata [123,124], Gentiana pannonica [123], Gentiana tibetica [123], Gentiana pneumonanthe and G. kurroo Royle [116,123,125,126,127], Gentiana davidii var. formosana (Hayata) [128], Gentiana straminea [103,129].…”

“…One of the recent reports [122] presented interesting results for the possibility to use recurrent somatic embryogenesis in long-term cultures of Gentiana lutea for production of synthetic Environmental Biotechnology -New Approaches and Prospective Applications seeds. After induction of somatic embryogenesis in the presence of auxins in the first cycle of in vitro cultures, recurrent somatic embryogenesis was performed in long-term cultures in the absence of phytohormones but in the presence of the sugar alcohols mannitol and sorbitol.…”

Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

“…In the case of G. lutea , somatic embryos were initiated by the epidermal cell divisions under osmotic stress treatment induced by mannitol. The results were confirmed with the help of histological analysis (Holobiuc and Câtanâ, 2012; Holobiuc, 2015).…”

Pedo-climatic and land use preferences of Gentiana lutea subsp. lutea in central Italy

“…Desmaret & Derchue 1988, Giovanardi & Barbaro 2009 and experienced in-vitro propagation (Holobiuc & Catana 2012, Petrova et al 2006. Contrariwise, in the Italian Apennines the effort to achieve these goals did not give satisfactory results.…”