Journal of Forestry Research

, Volume 30, Issue 1, pp 77–86 | Cite as

Preservation of Juniperus thurifera L.: a rare endangered species in Algeria through in vitro regeneration

  • Nadia Khater
  • Halima BenbouzaEmail author
Original Paper


Juniperus thurifera L. is an endemic Cupressaceae from the Aurès Mountains of north eastern Algeria and endangered, in part, due to the scarcity of viable seeds. It is threatened by other abiotic factors and the lack of an effective management strategy will increase its risk of extinction. The dearth of information on its in vitro regeneration impedes its application in forest management programs. We therefore developed a micropropagation protocol using microcuttings with auxiliary buds. Cuttings were grown on different combinations of media supplemented with plant growth regulators at different concentrations. The highest number of shoots and branches regenerated from original shoots was obtained on Woody Plant Medium (WPM) supplemented with 6-benzylaminopurine (BAP) (0.5 mg L−1) and 2,4-dichlorophenoxyacetic acid (2,4-D) (0.25 mg L−1). The best elongation of shoots was achieved with WPM supplemented with 0.5 mg L−1 of BAP and 0.25 or 1 mg L−1 of 2,4-D. On the second subculture, shoots had a higher number of branches than those of the first. The highest rooting rate, 38.8%, was obtained with shoots cultured in ½ Murashige and Skoog (MS) medium supplemented with 5.0 mg L−1 each of indol-3-butyric (IBA) and naphthalene acetic acid (NAA). Similarly, the highest root numbers and lengths were produced on ½ MS medium supplemented with IBA and NAA (5.0 mg L−1 each). During transfer to acclimatization, rates of plant losses of 50% occurred. The second part of the experiment showed that the best shoot callusing was on WPM supplemented with BAP and 2,4-D, with either the combination 0.5 + 0.25 or 0.25 + 0.25 mg L−1. The results of this research provide a starting point for further studies on in vitro regeneration of J. thurifera for the sustainable management of its unique ecosystem in the Mediterranean basin.


Juniperus thurifera Callus Conservation Regeneration In vitro culture 



We are most grateful to the lab staff of the C.R.Bt. (National Research Center of Biotechnology), namely Mrs. Khaoula Cherouana, Imen Benchiekh El Hocine, Ibtissem Benbelkacem and Abla Daas for technical assistance, and to Dr. Boualam Harfi for his support. Thanks are also due to Mr. Esadak Bouhrik, the Chief District of T’koutand and Mr. Atman Briki, the chief of Forestry Protection Service, for their valuable help with sample collections and field-tests of in vitro plants.


  1. Baravardi H, Ranjbar GA, Kamali S, Abadi F (2014) Investigation of the effects of growth regulators on callus induction in Juniperus excelsa L. Bull Env Pharmacol Life Sci 4:80–84Google Scholar
  2. Bonga JM, Klimaszewska KK, Von Aderkas P (2010) Recalcitrance in clonal propagation in particular conifers. Plant Cell Tissue Organ Cult 100:241–254CrossRefGoogle Scholar
  3. Campanoni P, Nick P (2005) Auxin-dependent cell division and cell elongation. 1-naphthaleneacetic acid and 2,4-dichlorophenoxyacetic acid activate different pathways. Physiol Plant 137:939–948CrossRefGoogle Scholar
  4. Castro MR, Belo AF, Afonso A, Zavatieri MA (2011) Micropropagation of Juniperus navicularis, an endemic and rare species from Portugal’s SW coast. Plant Growth Regul 65:223CrossRefGoogle Scholar
  5. Chiro L, Blanc CP (1997) Analysis of the distribution of reptiles in the massif of Aures, Algeria. Ecologi 28(4):281–292Google Scholar
  6. Chu CC, Wang CC, Sun CS, Msu C, Yin KC, Chu CY, Bi FY (1975) Establishment of an efficient medium for anther cultures of rice through comparative experiments on nitrogen sources. Sci Sin 18:659–668Google Scholar
  7. Dunberg A, Hsihan S, Sandberg G (1981) Auxin dynamic and the rooting cuttings of Pinus sylvestris. Physiol Plant 67:5–11CrossRefGoogle Scholar
  8. Ferradous A, Alifriqui M, Hafidi M, Duponnois R (2013) Essais de régénération artificielle du Genévrier thurifere (Juniperus thurifera L.). Ecol Mediterr 39(1):115–120Google Scholar
  9. Gauquelin T, Hassani MI, Lebreton P (1988) Le Genévrier thurifere, Juniperus thuriferaL. (Cupressacées): analyse biométrique et biochimique, position systématique. Ecol Mediterr 14:31–42Google Scholar
  10. Gauquelin T, Bertaudière V, Montes N, Badri W, Asmodé JF (1999) Endangered stands of thuriferous juniper in the western Mediterranean basin: ecological status, conservation and management. Biodivers Conserv 8:1479–1498CrossRefGoogle Scholar
  11. Ge X, Chu Z, Lin Y (2006) A tissue culture system for different germplasms of indica rice. Plant Cell Rep 25:392–402CrossRefGoogle Scholar
  12. Gómez MP, Segura J (1995) Axillary shoot proliferation in cultures of explants from mature Juniperus oxycedrus trees. Tree Physiol 15:625–628CrossRefGoogle Scholar
  13. Harry IS, Pulido CM, Thorpe TA (1995) Plantlet regeneration from mature embryos of Juniperus cedrus. Plant Cell Tissue Organ Cult 41:75–78CrossRefGoogle Scholar
  14. Hyndman SE, Hsegawa PM, Bressan RA (1982) Stimulation of root initiation from cultured rose shoots through the use of reduced concentrations of mineral soils. Hort Sci 17:82–83Google Scholar
  15. Jimenez MN, Navarro FB, Ripollmá, Bocioi DE, Simón E (2005) Effect of shelter tubes on establishment and growth of Juniperus thurifera L. (Cupressaceae) seedlings. Ann For Sci 62:717–725CrossRefGoogle Scholar
  16. Kocer ZA, Gozen AG, Onde S, Kaya Z (2011) Indirect organogenesis from bud explants of Juniperus communis L: effects of genotype, gender, sampling time and growth regulator. Dendrobiology 66:33–40Google Scholar
  17. Lloyd G, McCown B (1981) Commercially-feasible micropropagation of mountain laurel, kalmia latifolia by use of shoot-tip culture. Proc Intern Plant Prop Soc 30:421–427Google Scholar
  18. Loureiro J, Capelo A, Brito G, Rodriguez E, Silva S, Pinto G, Santos C (2007) Micropropagation of Juniperus phoenicea from adult plant explants and analysis of ploidy stability using flow cytometry. Biol Plant 51(1):7–14CrossRefGoogle Scholar
  19. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15(3):473–497CrossRefGoogle Scholar
  20. Negussie A (1997) in vitro induction of multiple buds in tissue culture of Juniper-us excelsa. For Ecol Manag 98:115–123CrossRefGoogle Scholar
  21. Oldfield S, Lusty C, MacKinven A (1998) The world list of threatened trees. World Conservation Press Cambridge, Cambridge, p 650Google Scholar
  22. Oliveira P, Barriga J, Cavaleiro C, Peixe A, Potes AZ (2003) Sustained in vitro root development obtained in Pinuspinea L. inoculated with ectomycorrhizal fungi. Forestry 76:579–587. CrossRefGoogle Scholar
  23. Ordas RJ, Rodriguez A, Rodriguez R, Sanchez R (1985) Desarrollo de técnicas de cultivo “in vitro” para la micropropagacion de variedades de manzanasidrera. Edafol Agrobactecnol 43:905–917Google Scholar
  24. Pritchard HW, Moat JF, Ferraz JBS, Marks TR, Luís J, Camargo C, Nadarajan J, Ferraz IDK (2014) Forest ecology and management innovative approaches to the preservation of forest trees. For Ecol Manag 333:88–98CrossRefGoogle Scholar
  25. Ragonezi C, Klimaszewska K, Castro MR, Lima M, Oliveira P, Zavattieri MA (2010) Adventitious rooting of conifers: influence of physical and chemical factors. Trees Struct Funct 24:975–992CrossRefGoogle Scholar
  26. Smith DR, Wareing PF (1972) The rooting of actively growing and dormant leafy cuttings in relation of endogenous hormone levels and photoperiod. New Phytol 71:483–500CrossRefGoogle Scholar
  27. Terrab A, Schönswetter P, Talavera S, Vela E, Stuessy TF (2008) Molecular phylogenetics and evolution range-wide phylogeography of Juniperus thurifera L., a presumptive keystone species of western Mediterranean vegetation during cold stages of the Pleistocene. Mol Phylogenet Evol 48(1):94–102CrossRefGoogle Scholar
  28. Trewavas AJ, Cleland RE (1983) Is plant development regulated by changes in concentration of growth substances or by changes in the sensitivity? Trends Biochem Sci 8:354–357CrossRefGoogle Scholar
  29. Zaidi MA, Khan S, Jahan N (2012) Micropropagation and conservation of three juniperus species (cupressaceae). Pak J Bot 44:301–304Google Scholar

Copyright information

© Northeast Forestry University and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Faculté SNVUniversité Batna 2BatnaAlgeria
  2. 2.Institut des Sciences Vétérinaires et Sciences AgronomiquesUniversité Batna 1BatnaAlgeria
  3. 3.Centre de Recherche en BiotechnologiesConstantineAlgeria

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