Skip to main content

Advertisement

Log in

Efficient protocol for in vitro propagation from bulb scale explants of Fritillaria ruthenica Wikstr. (Liliaceae), a rare ornamental species

  • Published:
Rendiconti Lincei. Scienze Fisiche e Naturali Aims and scope Submit manuscript

Abstract

An efficient protocol for in vitro propagation from bulb scale explants has been developed for Fritillaria ruthenica Wikstr., an endangered and rare species. The use of B5 medium supplemented with 5.0-µM 6-benzylaminopurine (BAP) and 2.0-µM α-naphthalene acetic acid (NAA) was found to be the most effective for adventitious bud induction and shoot multiplication (2.9 shoots per explant with 100% regeneration frequency). Adventitious regeneration occurred exclusively via direct organogenesis with the development of microshoots consisting of the rosette leaves, while bulb formation was not observed. The rooting microplants was carried out on a half-strength BDS hormone-free medium supplemented with activated charcoal at a concentration of 0.5 g/l resulted in 100% rhizogenesis with 5.2 roots per plantlet. Elevated sucrose concentrations (40.0 and 50.0 g/l) had no stimulating effect on bulblet differentiation at the in vitro rooting stage of F. ruthenica shoots. The regenerated plantlets were successfully transferred in a mixture of coconut fiber and sand (3:1) for acclimatization in greenhouse conditions with 72% of survival rate. The large bulbs developed only in ex vitro conditions at the end of the vegetation period.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Bacchetta L, Remotti PC, Bernardini C, Saccardo F (2003) Adventitious shoot regeneration from leaf explants and stem nodes of Lilium. Plant Cell Tissue Org Cult 74:37–44. https://doi.org/10.1023/A:1023315321931

    Article  CAS  Google Scholar 

  • Bardunov LV, Novikov VS (2008) Red Book of Russian Federation (plants and fungi). KMK Scientific Press Ltd., Moscow (In Russian)

    Google Scholar 

  • Carasso V, Mucciarelli M (2014) In vitro bulblet production and plant regeneration from immature embryos of Fritillaria tubiformis Gren. & Godr. Prop Ornament Plants 14(3):101–111

    Google Scholar 

  • Day PD, Berger M, Hill L, Fay MF, Leitch AR, Kelly LJ, Leitch IJ (2014) Evolutionary relationships in the medicinally important genus Fritillaria L. (Liliaceae). Mol Phylog Evol 80:11–19. https://doi.org/10.1016/j.ympev.2014.07.024

    Article  Google Scholar 

  • De Klerk GJ (2012) Micropropagation of bulbous crops: Technology and present state. Floriculture and ornamental biotechnology. Bulbous ornamentals, vol 6 (special iss 1). Global Science Books, London, pp 1–8

  • De Klerk GJ, Van Schadewijk KK, Gerrits M (1992) Growth of bulblets of Lilium speciosum in vitro and soil. Acta Hortic 325:513–520. https://doi.org/10.17660/ActaHortic.1992.325.71

    Article  Google Scholar 

  • Dunstan DJ, Short KC (1977) Improved growth of tissue cultures of the onion Allium cepa. Phisiol Plant 41(1):70–72

    Article  Google Scholar 

  • Erst AA, Erst AS (2011) In vitro propagation of rare plant Fritillaria dagana Turcz. ex Trautv. from bulb scales. Turcz 14(4):90–93 (In Russian with English abstract)

    Google Scholar 

  • Erst AA, Erst AS, Shaulo DN, Kulkhanova DS (2014) Conservation and propagation in vitro of rare species Fritillaria (Liliaceae). Flora Asian Russia 1(13):64–70 (In Russian with English abstract)

    Google Scholar 

  • Gamborg OL, Eveleigh DE (1968) Culture methods and detection of glucanases in cultures of wheat and barley. Can J Biochem 46(5):417–421

    Article  CAS  Google Scholar 

  • Gao SL, Zhu DN, Cai ZH, Jiang Y, Xu DR (1999) Organ culture of a precious Chinese medicinal plant—Fritillaria unibracteata. Plant Cell Tissue Org Cult 59:197–201

    Article  CAS  Google Scholar 

  • Jevremović S, Petrić M, Zivković S, Trifunović M, Subotić A (2010) Superoxide dismutase activity and isoenzyme profiles in bulbs of snake’s head fritillary in response to cold treatment. Arch Biol Sci 62(3):553–558. https://doi.org/10.2298/ABS1003553J

    Article  Google Scholar 

  • Kulkhanova DS, Erst AA, Novikova TI (2015) In vitro regeneration from bulbous scales of Fritillaria sonnikovae, an endemic species. Russ J Dev Biol 46(4):215–221. https://doi.org/10.1134/S1062360415040050

    Article  CAS  Google Scholar 

  • Langens-Gerrits MM, Miller WBM, Croes AF, De Klerk GJ (2003) Effect of low temperature on dormancy breaking and growth after planting in lily bulblets regenerated in vitro. Plant Growth Regul 40:267–275. https://doi.org/10.1023/A:1025018728178

    Article  CAS  Google Scholar 

  • Lin G, Li P, Li S-L, Chan S-W (2001) Chromatographic analysis of Fritillaria isosteroidal alkaloids, the active ingredients of Beimu, the antitussive traditional Chinese medicinal herb. J Chromatogr A 935:321–338

    Article  CAS  Google Scholar 

  • Liu X, Yang G (2012) Adventitious shoot regeneration of oriental lily (Lilium orientalis) and genetic stability evaluation based on ISSR marker variation. In Vitro Cell Dev Biol Plant 48:172–179. https://doi.org/10.1007/s11627-012-9429-0

    Article  Google Scholar 

  • Mohammadi-Dehcheshmeh M, Khalighi A, Naderi R, Sardari M, Ebrahimie E (2008) Petal: a reliable explant for direct bulblet regeneration of endangered wild populations of Fritillaria imperialis L. Acta Physiol Plant 30:395–399. https://doi.org/10.1007/s11738-007-0126-2

    Article  CAS  Google Scholar 

  • Muraseva DS, Novikova TI, Erst AA (2015) In vitro propagation and conservation of rare species Fritillaria meleagris L. from floral explants. Contemp Prob Ecol 8(6):754–763. https://doi.org/10.1134/s1995425515060128

    Article  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nasircilar AG, Mirici S, Karagüzel Ö, Eren Ö, Baktir İ (2011) In vitro propagation of endemic and endangered Muscari mirum from different explant types. Turk J Bot 35:37–43. https://doi.org/10.3906/bot-0907-90

    Article  CAS  Google Scholar 

  • Paek KY, Murthy HN (2002) High frequency of bulblet regeneration from bulb scale sections of Fritillaria thunbergii. Plant Cell Tissue Org Cult 68:47–252

    Article  Google Scholar 

  • Petrić M, Subotić A, Trifunović M, Jevremović S (2012) Morphogenesis in vitro of Fritillaria spp. In: Van Tuyl JM, Arens P (eds) Floriculture and ornamental biotechnology. Bulbous ornamentals, vol 6 (special iss 1). Global Science Books, London, pp 78–89

  • Petrić M, Subotić A, Jevremović S, Trifunović-Momčilov M, Tadić V, Grujić M, Vujčić Z (2015) Esterase and peroxidase isoforms in different stages of morphogenesis in Fritillaria meleagris L. in bulb-scale culture. C R Biol 338(12):793–802. https://doi.org/10.1016/j.crvi.2015.08.002

    Article  Google Scholar 

  • Podwyszyńska M (2012) The mechanisms of in vitro storage organ formation in ornamental geophytes. In: Van Tuyl JM, Arens P (eds) Floriculture and ornamental biotechnology. Bulbous ornamentals, vol 6 (special iss 1). Global Science Books, London, pp 9–23

  • Rix EM (2001) Fritillaria: a revised classification: together with an updated list of species. The Fritillaria Group of the Alpine Garden Society, London

    Google Scholar 

  • Rønsted N, Law S, Thornton H, Fay MF, Chase MW (2005) Molecular phylogenetic evidence for the monophyly of Fritillaria and Lilium (Liliaceae; Liliales) and the infrageneric classification of Fritillaria. Mol Phylogenet Evol 35:509–527. https://doi.org/10.1016/j.ympev.2004.12.023

    Article  CAS  Google Scholar 

  • Sedelnikova LL (2002) Biomorphology of geophytes in western Siberia. Nauka, Novosibirsk (In Russian)

    Google Scholar 

  • Seon JH, Paek KY, Gao WY, Park CH, Sung SN (1999) Factors affecting micropropagation of patogen-free stock in Fritillaria thunbergii. Acta Hortic 502:333–337

    Article  CAS  Google Scholar 

  • Takayama S, Misawa M (1979) Differentiation in Lilium bulbscales grown in vitro. Effects of various cultural conditions. Phisiol Plant 46:184–190. https://doi.org/10.1111/j.1399-3054.1979.tb06555.x

    Article  CAS  Google Scholar 

  • Teixeira da Silva JA, Dobránszki J (2016) Tissue culture of Muscari species: present achievements and future perspectives. Rend Fis Acc Lincei 27:427–441. https://doi.org/10.1007/s12210-016-0505-y

    Article  Google Scholar 

  • Xin G-Z, Lam Y-C, Maiwulanjiang M, Chan GKL, Zhu KY, Tang W-L, Ting-Xia Dong T, Shi Z-Q, Li P, Tsim KWK (2014) Authentication of bulbus Fritillariae cirrhosae by RAPD–derived DNA markers. Molecules 19:3450–3459. https://doi.org/10.3390/molecules19033450

    Article  CAS  Google Scholar 

  • Yin Z-F, Zhao B, Bi W-L, Chen L, Wang Q-C (2013) Direct shoot regeneration from basal leaf segments of Lilium and assessment of genetic stability in regenerants by ISSR and AFLP markers. In Vitro Cell Dev Biol Plant 49:333–342. https://doi.org/10.1007/s11627-013-9501-4

    Article  CAS  Google Scholar 

Download references

Acknowledgements

In our study, material from the collection of the Central Siberian Botanical Garden SB RAS—USU_440534 “Collection of living plants indoors and outdoors” was used.

Funding

The work was carried out with the financial support of the budgetary project of the Central Siberian Botanical Garden, SB RAS “Assessment of morphogenetic potential of North Asian plant populations by experimental methods” (no. AAAA-A17-117012610051-5) within the framework of the State Assignment.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Dinara S. Muraseva.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Muraseva, D.S., Novikova, T.I. Efficient protocol for in vitro propagation from bulb scale explants of Fritillaria ruthenica Wikstr. (Liliaceae), a rare ornamental species. Rend. Fis. Acc. Lincei 29, 491–497 (2018). https://doi.org/10.1007/s12210-018-0693-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12210-018-0693-8

Keywords

Navigation