Cytomorphological studies on somatic embryogenesis of Gentiana tibetica (King) and G. cruciata (L.)

Abstract

The process of plant regeneration via somatic embryogenesis of two gentianas, <em>Gentiana tibetica</em> and <em>G. cruciata</em> was described. For this purpose seedling explants were cultured on agar medium and later maintained in cell suspension. For callus initiation seedling explants like: cotyledons, hypocotyl and root were plated on a callus induction medium (CIM) composed of MS (1962), supplemented with 0.5 mg/l 2,4-D and 1.0 mg/l Kin. For the formation of cell suspension culture, embryogenic callus was transferred into liquid maintained medium (MM) composed of MS (1962), supplemented with 1.0 mg/l Dic + 0.1 mg/l NAA + 2.0 mg/l BAP + 80.0 mg/l SA. The conversion of somatic embryo into plantlets required a new medium (ECM) based on MS (1962) mineral salts, supplemented with 0.5 mg/l GA₃ + 1.0 mg/l Kin + 0.5 mg/l NAA. For cytomorphological studies of particular stages of embryogenesis, specimens were stained with dyes and reagents: 1. PAS reaction with leukofucsin, 2. Safranin + fast green, 3. Erlich's hematoxylin

Protoplast fusion in the genus Gentiana: genomic composition and genetic stability of somatic hybrids between Gentiana kurroo Royle and G. cruciata L.

“…For over 20 years, we have studied the morphogenic abilities of various gentian species in vitro, aimed at conserving and broadening their genetic diversity (Mikuła et al 1996(Mikuła et al , 2011Mikuła and Rybczyński 2001;Fiuk and Rybczyński 2008;Tomiczak et al 2016). The results of our experiments revealed the tremendous embryogenic potential possessed by Gentiana kurroo Royle.…”

Somatic Embryogenesis in Long-Term Cultures of Gentiana lutea L. in the Presence of Osmotic Stress

“…Similar results of SE development were reported in the case of different gentians species in the presence of dicamba, NAA, and BAP on agar and in liquid cultures. Suspensor-like structures were also described in globular somatic embryos of other gentians (Mikuła et al 1996(Mikuła et al , 2005c. In cultures of G. lutea, periodic transfer at 2-3 months is necessary owing to a significant increase in mass of the regenerative aggregates.…”

Polish Contribution to Global Research on Somatic Embryogenesis

“…e exploitation of embryogenic suspension cultures is shown in Figure 1. Studies on SE in gentians began in 1994 and resulted in the description of SE for G. cruciata and G. tibetica in two culture systems (i.e., on agar and in liquid medium) 2 years later (Mikuła et al, 1996). e embryogenic cell suspension system developed for these species was later applied to other gentians, namely, G. pannonica (Mikuła et al, 2002), G. kurroo (Fiuk & Rybczyński, 2008), and recently G. capitata and G. decumbens (Tomiczak & Markowski, 2021).…”