Changes of Gentiopicroside Synthesis during Somatic Embryogenesis inGentiana macrophylla

Abstract

IN VITRO plant regeneration of Gentiana macrophylla Pall. and determination of gentiopicroside content during somatic embryogenesis are described in the present work. The highest percentage of embryogenic callus formation was observed in Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg/L 6-benzylaminopurine (BA). Calli were subcultured on MS medium containing 1.0 mg/L 2,4-D, 1.0 mg/L BA and 500 mg/L lactalbumin hydrolysate (LH) at intervals of 25 days. A higher frequency of somatic embryo maturation was achieved on MS medium containing B5 vitamins (MB) supplemented with different concentrations of 1-naphthaleneacetic acid (NAA) and BA than with a combination of NAA and kinetin (KT). Addition of AgNO(3) improved maturation of somatic embryos while thidiazuron (TDZ) promoted vitrification. The gentiopicroside contents of embryogenic calli and globular-, heart-, torpedo-, and cotyledon-shaped embryoids were determined by high-performance liquid chromatography (HPLC). Gentiopicroside was not detectable in embryogenic calli, but in all types of somatic embryos. The highest gentiopicroside content was observed in cotyledon-shaped embryoids, reaching more than 12 mg/g dry weight.

The Gentianaceae - Volume 2: Biotechnology and Applications

“…Plants of G. punctata grown in vitro on MS solid medium supplemented with 0.25-2.0 mg/l BAP contained the highest amount of gentiopicrin and swertiamarin in the aerial parts. Nonetheless, in all the stages of somatic embryogenesis studied, these metabolites were detected with the highest concentrations in embryoids being in the cotyledonary stage (Chen et al 2009). For G. straminea, BAP treatment was more beneficial for gentiopicroside formation than NAA, IAA, or 2,4-D.…”

“…The similarity of both types of roots is due to their origin; they are adventitious roots formed at the epicotyl of the explant . Recently, an effective protocol for plant regeneration from hairy roots via indirect somatic embryogenesis was described for G. macrophylla (Wu et al 2011), by using a two-step procedure Rybczyński et al involving (1) the application of MS medium supplemented with 0.5-2.5 mg/l 2,4-D or 2,4-D + 0.5 mg/l BAP (Chen et al 2009) resulting in embryogenic callus formation and (2) the transfer to MS PGR-free medium which stimulates callus for the production of somatic embryos including cotyledonary stage embryo. The roots had a clearly defined stele, a pericycle that is not very distinct, an innermost endodermis, and cortical layer.…”

“…However, there are some species, namely G. pneumonanthe, G. kurroo, and G. macrophylla which showed somatic embryos regeneration potential on leaf blade (Bach and Pawlowska 2003;Chen et al 2009). Attempts by of other researches to initiate regeneration of adventitious shoots from leaf explants of the species mentioned as well as G. punctata were also unsuccessful .…”

In Vitro Manipulation and Propagation of Gentiana L. Species from the Ukrainian Flora

“…These results illustrated the dependence of G. lutea regeneration capacity on culture conditions, especially on PGRs. However, there are some species, namely G. pneumonanthe, G. kurroo, and G. macrophylla which showed somatic embryos regeneration potential on leaf blade (Bach and Pawlowska 2003;Fiuk and Rybczyński 2008;Chen et al 2009). …”

Chemical and Genetic Comparative Analysis of Gentiana crassicaulis and Gentiana macrophylla

“…Previous studies mainly focused on the analysis of G . macrophylla or G . crassicaulis , separately, through either chemical or genetic techniques.…”