Comprehensive Medicinal Plant Database

Tissue Culture Literation

Plant latin nameGentiana scabra Bungeļ¼ŒGentiana manshurica Kitagawa
Literature codeGentiana_scabra-Ref-1
ReferenceHosokawa K et al., Plant Cell Reports 15: 578-581 (1996)
SummarySeveral culture conditions were examined for promoting efficient plant regeneration from explants of Gentiana Adventitious shoot regeneration from leaf explants of cv.WSP-3 was very superior on MS medium, compared to B5medium, supplemented with four cytokinins (TDZ, 4PU-30,BA and zeatin). An auxin / cytokinin combination was required for regeneration. TDZ was the most effective cytokinin, while NAA was more effective than IAA or 2,4-D.Optimum conditions for regeneration from explants (leaf,stem and root) of cv. WSP-3, evaluated in terms of regeneration frequency and number of regenerated shoots per explant, were TDZ and NAA in combination, 5 - 10 mg/1 and 0.1 mg/l for leaf and stem explants, and 10 mg/l and 1 mg/l for root explants, respectively. Application of these conditions to eight other commercial CUIlivars resulted in 30-100% regeneration from leaf explants. The number of chromosomes in each of ten regenerated plants of each cultivar was diploid, 2n=26. Shoots regenerated in vitro were rooted in phytohormone-free medium and transferred to soil.
ObjectivesAdventitious shoot regeneration from leaf, stem and root explants
MaterialsSeeds of five Gentiana triflora cultivars (F1 hybrid) Homoi, Ihatovo, Iwate, lwate-otome and Maciry; an interspecific hybrid cultivar (FI hybrid, G. triJlora x G. scabra ) Albireo; and three in vitro shoot cultures of interspecific hybrid cultivars (clonal variety, G. triflora x G. scabra) H-3, Polarno white and WSP-3, were obtained from Iwate Horticullural Experiment Station..
ExplantLeaf, stem, root
Initial cultureSeeds were surface-disinfected with 1% sodium hypochlorite solution for 5 min followed by two rinsings in sterilized distilled water. The disinfected seeds were germinated on MS medium supplemented with 30 g/l sucrose and 2 g/l gellan gum. All axillary buds were cultured on the same medium. Plantlets from the seedlings or in vitro shoot cultures were routinely subcultured at 200C for 16 h daytime under fluorescent lamps (50 /z mol m -2 s-l). Subcultures were made once every two
Shoot multiplicationAdventitious shoot regeneration from leaf explants of cv.WSP-3 was more frequent on MS medium than B5 medium in all cases. TDZ (1 - 10 rag/l) in combination with NAA (0.1 mg/1) proved to be optimal for regeneration. Regeneration from stem and root explants was greatest at 5-10 mg/1 TDZ in combination with 0.1 mg/1 NAA, and at 10 mg/1 TDZ in combination with 1 rag/1 NAA, respectively
RootingShoots produced in culture were easily rooted in phytohormone-free medium in four weeks, and then acclimatized
AcclimationShoots produced in culture were easily rooted in phytohormone-free medium in four weeks, and then acclimatized
Planting
Cultivation conditionsIn greenhouse
Traints of regenerantsEach of twenty regenerated plants grown in a greenhouse were phenotypically normal with respect to leaf shape and growth features during early growth stages
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Extraction
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