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  Eur.J.Hortic.Sci. 82 (1) 54-60 | DOI: 10.17660/eJHS.2017/82.1.6
ISSN 1611-4426 print and 1611-4434 online | © ISHS 2017 | European Journal of Horticultural Science | Original article

In vitro propagation of medicinal and ornamental plant Lysimachia davurica

Yanni Zhang, Dongling Sun and Shangchun Hu
College of Landscape Architecture, Northeast Forestry University, Harbin 150040, China

Lysimachia davurica is an important medicinal and ornamental plant species. An efficient in vitro regeneration system on excised leaves of L. davurica was developed. The experimental conditions of tissue culture system were investigated based on the effect of different concentrations and combinations of plant growth regulators (PGRs), explant orientations, leaf segments, and concentrations of activated charcoal. Two types of callus induction medium using Murashige and Skoog (MS) medium + 1.0 mg L-1 NAA + 0.5 mg L-1 6-BA and 1.0 mg L-1 NAA + 1.0 mg L-1 6-BA had a callus inducing rate of 95.56%. The shortest time to callus appearance was 7 d. Proximal segments were better for callus induction than middle and distal ones. Placing leaf with abaxial side down on medium was better than adaxial side for inoculation. For the combinations of 6-BA and NAA tested, MS + 0.5 mg L-1 6-BA and 0.1 mg L-1 NAA was best for callus proliferation, while MS + 1.0 mg L-1 6-BA and 0.1 mg L-1 NAA was optimal for callus differentiation and shooting. For the combinations of PGRs tested, the best rooting medium is 1/2 MS + 0.5 mg L-1 IBA + 0 mg L-1 NAA, with 99.89% rooting rate. On plantlets rooting, 1 g L-1 activated charcoal performed better than others (0, 2, 3, and 4 g L-1). The optimal transplanting medium was using vermiculite and humus as substrate in the proportion of 2:1 (v/v), with 97.78% survival rate. This protocol could be an efficient method for the micropropagation of L. davurica.

Keywords micropropagation, tissue culture, leaf, shoot regeneration, callus

Significance of this study

What is already known on this subject?

  • Micropropagation of L. davurica had been achieved using hypocotyl and cotyledon stem section as explants while its efficiency of plant regeneration is relatively low. To date, a tissue culture protocol for regeneration of L. davurica using leaf tissue explants has not been reported.
What are the new findings?
  • Our study provides a tissue culture protocol for regeneration of L. davurica using leaf tissue explants, based on the effect of different hormone concentrations and combinations, explant orientations, leaf segments, and concentrations of activated charcoal.
What is the expected impact on horticulture?
  • The results could help promote rapid production of L. davurica for growers. The in vitro plantlets may be suitable to set up a transformation system to facilitate the production of transgenic plants with desirable horticultural traits or medicinal contents.

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  • Ahmad, N., Faisal, M., Anis, M., and Aref, I.M. (2010). In vitro callus induction and plant regeneration from leaf explants of Ruta graveolens L. S. Afr. J. Bot. 76, 597–600.

  • Aina, O.O., Quesenberry, K.H., and Gallo, M. (2015). Culture vessel and auxin treatments affect in vitro, rooting and ex vitro, survival of six Arachis paraguariensis genotypes. Sci. Hortic. 183, 167–171.

  • Cao, Z., Sui, S., Yang, Q., and Deng, D. (2016). Somaclonal variation in ‘Red Flash’ caladium: morphological, cytological and molecular characterization. Plant Cell Tissue Org. Cult. 126, 269–279.

  • Cardoso, J.C., and Habermann, G. (2014). Adventitious shoot induction from leaf segments in Anthurium andreanum is affected by age of explant, leaf orientation and plant growth regulator. Hortic. Environ. Biotechnol. 55(1), 56–62.

  • Chen, X. (2012). Tissue culture rooting inducement experiment of Acacia cincinata and Acacia crassicarpa. J. Fujian For. Sci. Tech. 39, 108–110, 119.

  • Cheng, C., Zang, G., Zhao, L., Gao, C., Tang, Q., Chen, J., Guo, X., Peng, D., and Su, J. (2016). A rapid shoot regeneration protocol from the cotyledons of hemp (Cannabis sativa L.). Ind. Crops Prod. 83, 61–65.

  • Cheruvathur, M.K., Abraham, J., and Thomas, T.D. (2013). Plant regeneration through callus organogenesis and true-to-type conformity of plants by RAPD analysis in Desmodium gangeticum (Linn.) DC. Appl. Biochem. Biotechnol. 169, 1799–1810.

  • Debnath, S.C. (2009). A two-step procedure for adventitious shoot regeneration on excised leaves of lowbush blueberry. In Vitro Cell. Dev. Biol.: Plant. 45(2), 122–128.

  • Kakani, A., Li, G., and Peng, Z. (2009). Role of aux1 in the control of organ identity during in vitro organogenesis and in mediating tissue specific auxin and cytokinin interaction in Arabidopsis. Planta. 229(3), 645–657.

  • Karami, O., Piri, K., and Bahmani, R. (2009). Plant regeneration through callus cultures derived from immature-cotyledon explants of oleaster (Elaeagnus angustifolia L.). Trees-Struct. Funct. 23(2), 335–338.

  • Kim, K.M., Min, Y.K., Yun, P.Y., Chandrasekhar, T., Lee, H.Y., and Song, P.S. (2007). Production of multiple shoots and plant regeneration from leaf segments of fig tree (Ficus carica L.). J. Plant Biol. 50(4), 440–446.

  • Liu, H.Z., Tan, X.H., and Zheng, T. (2008). Establishment of regeneration systems of Pyrus ussuriensis leaves. J. Jilin Agric. Univ. 30(4), 472–476.

  • Mazumdar, P., Basu, A., Paul, A., Mahanta, C., and Sahoo, L. (2010). Age and orientation of the cotyledonary leaf explants determine the efficiency of de novo plant regeneration and Agrobacterium tumefaciens-mediated transformation in Jatropha curcas L. S. Afr. J. Bot. 76(2), 337–344.

  • Mendes, A.F.S., Cidade, L.C., Otoni, W.C., Soaresfilho, W.S., and Costa, M.G.C. (2011). Role of auxins, polyamines and ethylene in root formation and growth in sweet orange. Biol. Plantarum 55(2), 375–378.

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

  • Murthy, K.S.R., Kondamudi, R., and Vijayalakshmi, V. (2010). Micropropagation of an endangered medicinal plant Ceropegia spiralis L. J. of Agric. Tech. 6(1), 179–191.

  • Phulwaria, M., Shekhawat, N.S., Rathore, J.S., and Singh, R.P. (2013). An efficient in vitro regeneration and ex vitro rooting of Ceropegia bulbosa Roxb. – A threatened and pharmaceutical important plant of Indian Thar Desert. Ind. Crops Prod. 42(3), 25–29.

  • Prammanee, S., Thumjamras, S., Chiemsombat, P., and Pipattanawong, N. (2011). Efficient shoot regeneration from direct apical meristem tissue to produce virus-free purple passion fruit plants. Crop Prot. 30(11), 1425–1429.

  • Qu, C., Xi, T., and Miao, L. (1990). Tissue culture and micro-propagation of Lysimachia vulgaris var. davarica. Plant Physiol. Commun. 2, 42.

  • Tallσn, C.I., Porras, I., and Pιrez-Tornero, O. (2012). Efficient propagation and rooting of three citrus rootstocks using different plant growth regulators. In Vitro Cell. Dev. Biol.: Plant. 48(5), 488–499.

  • Thomas, T.D. (2008). The role of activated charcoal in plant tissue culture. Biotechnol. Adv. 26(6), 618–631.

  • Thomas, T.D., and Hoshino, Y. (2015). Callus induction, high frequency shoot organogenesis and assessment of clonal fidelity in Torenia bicolor Dalzell. J. Appl. Res. Med. Aromat. Plants. 2(4), 188–194.

  • Tian, J., Zou, Z., Xu, L., and Yang, S. (2005). Studies on chemical constituents (triterpenoid saponin) of Lysimachia davuria. Chinese Pharm. J. 40, 1133–1134.

  • Turker, A.U., and Guner, B. (2013). Efficient plant regeneration of yellow loosestrife (l.), a medicinal plant. Acta Biol. Hungarica 64(2), 218–230.

  • U.S. National Plant Germplasm System. (2016). Taxon: Lysimachia davurica Ledeb. Available at (accessed July 16, 2016).

  • Zhang, Y., and Han, R. (2012). Tissue culture and rapid propagation of Orostachys malacophyllus leaves with axillary buds. Pratacultural Sci. 29(5), 735–740.

  • Zhang, Y., Sun, D., and He, M. (2013). Optimum germination conditions of Lysimachia davurica seeds. J. Northeast For. Univ. 42, 27–29.

  • Zheng, W., Xu, X., Dai, H., and Chen, L. (2009). Direct regeneration of plants derived from in vitro, cultured shoot tips and leaves of three Lysimachia species. Sci. Hortic. 122(1), 138–141.

Received: 10 October 2016 | Accepted: 12 January 2017 | Published: 23 February 2017 | Available online: 23 February 2017

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