Cover Image

Somatic embryogenesis and plant regeneration from cell suspension cultures of Gentiana kurroo Royle.

Shiwani Kaushal, Arushdeep Sidana


Gentiana kurroo Royle, native to North- Western Himalayas, is one of the critically endangered perennial herbs, which is being overexploited due to its multiple medicinal uses. The goal of this work was to develop a protocol for somatic embryogenesis and efficient plant regeneration through cell suspension culture. Selected ranges of plant growth regulators were experimented for somatic embryogenesis. Light micrographs of the established suspension cell cultures of somatic embryos at different developmental stages were also observed under light microscope. Somatic embryogenesis was induced from the leaf explants and the maximum induction frequency (97.2%) was obtained on Murashige and Skoog (MS) medium supplemented with 0.5 mg/l each of NAA, BAP and TDZ. Cell suspension cultures were established in MS liquid medium containing IAA (0.5 mg/l) + BAP (1.0 mg/l) with 89-93% viable cells. Transfer of cotyledonary somatic embryos to the MS agar medium containing 2, 4-D (0.4 mg/l) and KN (1.5 mg/l) resulted in somatic embryogenesis at high frequencies (80%) with an average of 28 ± 0.2 embryos/ callus piece. After transfer onto the half strength MS medium without growth regulators approximately 80-90% somatic embryos developed into complete plantlets. The protocol described here could be used for somatic hybridization, genetic transformation, isolation of protoplasts and large-scale propagation of G. kurroo.


Cell suspension, Gentiana kurroo, Murashige and Skoog (MS), Somatic embryogenesis

Full Text:



Aboshama HMS, Somatic embryogenesis proliferation, maturation and germination in Cajanus cajan, World Journal of Agricultural Science, 2011, 7, 86-95.

Bach A, Pawłowska B, Somatic embryogenesis in Gentiana pneumonantheL, Acta Biologica Cracoviensia, 2003, 45, 79-86.

Chowdhury P, Das M, Sikdar SR, Pal A, Influence of the physiological age and position of the nodal explants on micropropagation of field-grown Dendrocalamus strictus, Plant Cell Biotechnology and Molecular Biology, 2004, 5, 45-50.

Devi WS, Sharma GJ, In vitro propagation of Arundinaria callosa Munro—an edible bamboo from nodal explants of mature plants, Open Plant Science Journal, 2009, 3, 35-39.

Eapen S, George L, High frequency plant regeneration through somatic embryogenesis in finger millet (Eleusina carcana Gaertn), Plant Science, 1989, 61, 127-130.

Fiuk A, Rajkiewicz M, Rybczynski JJ, In vitro culture of Gentiana kurroo (Royle). Biotechnologia, 2003, 3, 267-274.

Fiuk A, Rybczynski JJ, The effect of several factors on somatic embryogenesis and plant regeneration in protoplast cultures of Gentiana kurroo (Royle), Plant Cell Tissue and Organ Culture, 2007, 91, 263-271.

Fiuk A, Rybczynski JJ, Factors influencing efficiency of somatic embryogenesis of Gentiana kurroo (Royle) suspension, Plant Biotechnology Reports, 2008 a, 1, 33-39.

Fiuk A, Rybczynski JJ, Genotype and plant growth regulator dependent response of somatic embryogenesis from Gentiana spp. leaf explants, In Vitro Cellular and Developmental Biology, 2008 b, 44, 90-99.

Fiuk A, Rybczynski JJ, Morphogenic capability of Gentiana kurroo Royle seedling and leaf explants, Acta Physiologiae Plantarum, 2008 c, 30, 157-166.

Hatanaka T, Arakawa O, Yasuda T, Uchida N, Yamaguchi T, Effect of plant growth regulators on somatic embryogenesis in leaf culture of Coffea canephora, Plant Cell Reports, 1991, 10, 179-182.

Kaushal S, Sidana A, Dev K, In vitro plant production through apical meristem culture of Gentiana kurroo Royle, Journal of Medicinal Plants Studies, 2014, 3, 04-09.

Luis PBC, Adriane CMGM, Silvica BRCC, Ana Christina MB, Plant regeneration from seedlings explants of Eucalyptus grandisEurophylla, Plant Cell Tissue and Organ Culture, 1999, 56, 17-23.

Luo JP, Jia JF, Callus induction and plant regeneration from hypocotyl explants of the forage legume Astragalus adsurgens, Plant Cell Reports, 1998, 17, 567-570.

Mikuła A, Tykarska T, Kuras M, Iwanowska A, Rybczynski JJ, Studies on the morphogenic potential of Gentians in cell suspension, In: Beitrage zur Zuchtungsforschung, Symposium Breeding Research on Medicinal and Aromatic Plants, Quedlinburg, Germany, 1996, 290-295.

Mikuła A, Rybczynski JJ, Somatic embryogenesis of Gentiana genus I, the effect of the preculture treatment and primary explant origin on somatic embryogenesis of Gentiana cruciata (L.), G. pannonica (Scop.), and G. tibetica (King), Acta. Physiologiae Plantarum, 2001, 23, 15-25.

Mikuła A, Rybczynski JJ, Skierski J, Latkowska MJ, Fiuk A, Somatic embryogenesis of Gentiana genus IV, characterization of Gentiana cruciata and Gentiana tibetica embryogenic cell suspensions, In: Hvoslef-Eide, A K, Preil W, (Eds), Liquid culture systems for plant propagation: Springer, Dordrecht, 2005, 345-358.

Murashige, T, Skoog F, A revised medium for rapid growth and bioassays with tobacco tissue culture, Physiologia Plantarum, 1962, 15, 473–497.

Nagesh KS, Shanthamma C, Pullaiah T, Somatic embryogenesis and plant regeneration from callus cultures of Curculigo orchioides Gaertn, Indian Journal of Biotechnology, 2010, 9, 408-.413.

Raina R, Behera MC, Chand R, Sharma Y, Reproductive Biology of Gentiana kurroo Royle, Current Science, 2003, 85, 667-670.

Sharma N, Chandel KPS, Paul A, In vitro propagation of Gentiana kurroo – an indigenous threatened plant of medicinal importance, Plant Cell Tissue and Organ Culture, 1993, 34, 307-309.

Sharma A, Kaur R, Sharma N, In vitro morphogenic response of different explants of Gentiana kurroo Royle from Western Himalayas-an endangered medicinal plant, Physiology and Molecular Biology of Plants, 2014, 20,249-256.

Tiwari V, Singh BD, Tiwari NK, Shoot regeneration and somatic embryogenesis from different explants of Brahmi (Bacopa monnieri), Plant Cell Reports, 1998, 17, 538-543.

Yemets AI, Klimkina LAA, Tarassenko LV, Blume YB, Efficient callus formation and plant regeneration of goosegrass (Eleusine indica), Plant Cell Reports, 2003, 21, 503-510.

Zoriniants SE, Nosov AV, Gonzalez MM, Zeel MM, Vargas VML, Variation of nuclear DNA content during somatic embryogenesis and plant regeneration of Coffea arabica using cytophotometry, Plant Science, 2003, 164, 141-146.


Copyright (c) 2018 Annals of Plant Sciences

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.