Journal of Forestry Research

, Volume 22, Issue 1, pp 47–52 | Cite as

Influence of cytokinins, basal media and pH on adventitious shoot regeneration from excised root cultures of Albizia lebbeck

  • Shahnaz Perveen
  • Ankita Varshney
  • Mohammad AnisEmail author
  • Ibrahim M. Aref
Original Paper


A highly reproducible and efficient in vitro shoot regeneration system was developed in a potential medicinal plant, Albizia lebbeck using root explants. Root explants from 15 day-old-aseptic seedlings were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations (0.5, 2.5, 5.0, 7.5 and 10.0 μM) of 6-Benzyladenine (BA), Kinetin (Kn), 2-Isopentenyl adenine (2-iP) singly as well as in combination with α-Naphthalene acetic acid (NAA) (0.1, 0.5, 1.0, 1.5 and 2.0 μM). The highest rate of shoot multiplication (16.0 ± 1.87 for the average shoot number and 5.16 ± 0.38 cm for shoot length) was achieved on MS medium supplemented with 7.5 μM BA and 0.5 μM NAA. The effects of medium type, medium strength, pH and subculture on shoot induction and proliferation were also tested. An average of 21.6±2.87 shoots per explants could be obtained following this protocol. Rooting was achieved on microshoots using half strength MS medium with 2.0 μM Indole-3-butyric acid (IBA) after four weeks of culture. The in vitro raised healthy plantlets were successfully established in earthen pots containing garden soil and grown in greenhouse with >80% survival rate.


Albizia lebbeck direct organogenesis fabaceae plant regeneration root explants 


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  1. Ahmad N, Wali SA, Anis M. 2008. In vitro production of true-to-type plants of Vitex negundo L. from nodal explants. Journal of Horticultural Science and Biotechnology, 83(3): 313–317.Google Scholar
  2. Anis M, Varshney A, Siddique I. 2010. In vitro clonal propagation of Balanites aegyptiaca (L.) Del. Agroforestry Systems, 78: 151–158.CrossRefGoogle Scholar
  3. Basu P, Chand S. 1996. Regeneration of plantlets from root derived callus of Egyption henbane. Cell Chromosome Research, 19: 31–34.Google Scholar
  4. Bhat SR, Chitralekha P, Chandel KPS. 1992. Regeneration of plants from long term root culture of lime, Citrus aurantifolia (Christm.) Swing. Plant Cell Tissue and Organ Culture, 29: 19–25.CrossRefGoogle Scholar
  5. Bhuyan AK, Pattnaik S, Chand PK. 1997. Micropropagation of Curry Leaf Tree [Murraya koenigii (L.) Spring.] by axillary proliferation using intact seedlings. Plant Cell Reports, 16: 779–782.CrossRefGoogle Scholar
  6. Chand PK, Sahoo Y, Pattnaik SK, Pattnaik SN. 1995. In vitro meristem culture — an efficient ex situ conservation strategy for elite mulberry germplasm. In: Mohanty RC (ed), Environment: change and management, Kamla Raj Enterprises, New Delhi, India, pp 127–133.Google Scholar
  7. Chaudhari KN, Ghosh S, Jha S. 2004. The root: A potential source of competent cells for high frequency regeneration in Tylophora indica. Plant Cell Reports, 22: 731–740.CrossRefGoogle Scholar
  8. Chintawar SD, Somani RS, Kasture VS, Kasture SB. 2002. Nootropic activity of Albizia lebbeck in mice. Journal of.Ethnopharmacology, 81(3): 299–305.CrossRefPubMedGoogle Scholar
  9. Dixit AK, Mishra LN. 1997. Macrocyclic budmunchiamine alkaloids from Albizia lebbeck. Journal of Natural Products, 60(10): 1036–1037.CrossRefGoogle Scholar
  10. Duncan DB. 1955. Multiple range and multiple F tests. Biometrics, 11:1–42.CrossRefGoogle Scholar
  11. El-Mousallamy AMD. 1998. Leaf flavonoids of Albizia lebbeck. Phytochemistry, 48(4): 759–761.CrossRefPubMedGoogle Scholar
  12. Faisal M, Siddique I, Anis M. 2006. An efficient plant regeneration system for Mucuna pruriens L. (DC.) using cotyledonary node explants. In Vitro Cellular and Developmental Biology-Plant, 42: 59–64.CrossRefGoogle Scholar
  13. Gamborg OL, Miller RA, Ojima K. 1968. Nutrient requirements of suspention cultures of soyabean root cells. Experimental Cell Research, 50: 151–158.CrossRefPubMedGoogle Scholar
  14. Gautam VK, Nanda N, Gupta SC. 1993. Development of shoots and roots in anther derived callus of Azadirachta indica A. Juss-A medicinal tree. Plant Cell Tissue and Organ Culture, 34: 13–18.CrossRefGoogle Scholar
  15. George EF, Hall MA, De Klerk GJ. 2008. Plant propagation by tissue culture. Vol 1: The background, 3rd edn. Springer, Dordrecht.Google Scholar
  16. Georges D, Chemienx JC, Ochatt SD. 1993. Plant regeneration from aged callus of woody ornamental species Lonicera japonica cv. Hell’s prolific. Plant Cell Reports, 13: 91–94.CrossRefGoogle Scholar
  17. Gharyal PK, Maheshwari SC. 1983. In vitro differentiation of plantlets from tissue cultures of Albizia lebbeck L. Plant Cell Tissue and Organ Culture, 2: 49–53.CrossRefGoogle Scholar
  18. Gharyal PK, Maheshwari SC. 1990. Differentiation in explants from mature leguminous trees. Plant Cell Reports, 8: 550–553.CrossRefGoogle Scholar
  19. Gupta RS, Chaudhary R, Yadav RK, Verma SK, Dobhal MP. 2005. Effect of saponins of Albizia lebbeck Benth. Bark on the reproductive system of male albino rats. Journal of Ethnopharmacology, 96(1–2): 31–36.CrossRefPubMedGoogle Scholar
  20. Hiregoudar LV, Murthy HN, Bhat JG, Nayeem A, Hema BP, Hahn EJ. 2006. Rapid clonal propagation of Vitex trifolia. Biologia Plantarum, 50: 291–295.CrossRefGoogle Scholar
  21. Husain MK, Anis M, Shahzad A. 2008. In vitro propagation of a multipurpose leguminous tree (Pterocarpus marsupium roxb.) using nodal explants. Acta Physiologia Plantarum, 30: 353–359.CrossRefGoogle Scholar
  22. Husain MK, Anis M. 2004. In vitro proliferation of Shoots of Melia azedarach L. from mature trees. In: D’sauza L, Anutadha M, Nivas S, Hedge S, Ragendra K (eds) Biotechnology For A Better Future, SAC Publications, Manglore, India. pp 294–301.Google Scholar
  23. Hussain TM, Chandrasekhar T, Gopal GR. 2008. Micropropagation of Sterculia urens Roxb, an endangered tree species from intact seedlings. African Journal of Biotechnology, 7(2): 95–101.Google Scholar
  24. Kanthrajah AS, Rechard GD, Dodd WA. 1992. Roots as source of explants for successful micropropagation of Cacambola (Averrhoa carambola). Scientia Horticulturae, 51: 169–177.CrossRefGoogle Scholar
  25. Karim MZ, Yokota S, Rahman MM, Eizawa J, Saito Y, Azad MAK, Ishiguri F, Iizuka K, Yoshizawa N. 2007. Efficient adventitious shoot regeneration from root explant of Aralia elata Seem. International Journal of Botany, 3(4): 390–393.CrossRefGoogle Scholar
  26. Karim MZ, Yokota S, Rahman MM, Eizawa J, Saito Y, Azad MAK, Ishiguri F, Iizuka K, Yoshizawa N. 2007. Effect of the sucrose concentration and pH level on shoot regeneration from callus in Araria elata Seem. Asian Journal of Plant Science, 6(4): 715–717.CrossRefGoogle Scholar
  27. Kim KH, Park HK, Park MS, Yeo UD. 2001. Effects of auxin and cytokinin on organogenesis of soybean Glycine max L. Journal of Plant Biotechnology, 3: 95–100.Google Scholar
  28. Lloyd GB, McCown BH. 1980. Commercially feasible micropropagation of mountain laurel (Kalmia latifolia) by use of shoot tip culture. International Plant Propagators Society Comb Proc, 30: 421–437.Google Scholar
  29. Maa YT, Hsiaob SC, Chenb HF, Hsu FL. 1997. Tannins from Albizia lebbeck. Phytochemistry, 46(8): 1451–1452.CrossRefGoogle Scholar
  30. Mamun ANK, Matin MN, Bari MA, Siddique NA, Sultana RS, Rahman MH, Musa ASM. 2004. Micropropagation of woody legume (Albizia lebbeck) through tissue culture. Pakistan Journal of Biological Science, 7(7): 1099–1103.CrossRefGoogle Scholar
  31. Martin KP. 2004. Plant regeneration protocol of medicinally important Andrographis paniculata (Burm. F.) Wallich ex Nees via somatic embryogeneses. In Vitro Cellular and Developmental Biology-Plant, 40: 204–209.CrossRefGoogle Scholar
  32. Martinez-Pulido C, Harry IS, Thorpe TA. 1992. Optimization of bud induction in cotyledonary explants of Pinus canariensis. Plant Cell Tissue and Organ Culture, 29: 247.CrossRefGoogle Scholar
  33. Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiologia Plantarum, 15: 473–497.CrossRefGoogle Scholar
  34. Nair LG, Seeni S. 2003. In vitro multiplication of Calophyllum apitalum (Clusiaceae) an endemic medicinal tree of the Western Ghats. Plant Cell Tissue and Organ Culture, 75: 169–174.CrossRefGoogle Scholar
  35. Ovecka M, Bobak M, Samaj J. 2000. A comparative structure analysis of direct and indirect shoot regeneration of Papaver somniferum L. in vitro. Journal of Plant Physiology, 157: 281–289.Google Scholar
  36. Pratibha N, Saxena VS, Amit A, D’sauza P, Bagchi M, Bagchi D. 2004. Antiinflammatory activities of Aller-7, a novel polyherbal formulation for allergic rhinitis. International Journal of Tissue Reactions, 26(1–2): 43–51.PubMedGoogle Scholar
  37. Saha A, Ahmed M. 2009. The analgesic and anti-inflammatory activities of the extract of Albizia lebbeck in Animal Model. Pakistan Journal of Pharmaceutical Science, 22(1): 74–77.Google Scholar
  38. Sakano K. 1990. Proton/Phosphate steichiometry in uptake of inorganic phosphate by cultured cells of Catharanthus roseus (L.) G. Don. Plant Physiology, 93: 479–483.CrossRefPubMedGoogle Scholar
  39. Schmitz U, Lorz H. 1990. Nutrient uptake in suspention cultures of gramineae. 2. Suspension culture of rice (Oryza sativa L.). Plant Science, 66: 95–111.CrossRefGoogle Scholar
  40. Scholten HJ, Pierik RLM. 1998. Agar as a gelling agent: chemical and physical analysis. Plant Cell Reports, 17: 230–235.CrossRefGoogle Scholar
  41. Shahin-uz-zaman M, Ashrafuzzaman M, Shahidul HM, Nahar LL. 2008. In vitro clonal propagation of the Neem tree (Azadirachta indica A. Juss.). African Journal of Biotechnology, 7(4): 385–391.Google Scholar
  42. Siddique I, Anis M. 2009. Direct plant regeneration from nodal explants of Balanites aegyptiaca L. (Del.) a valuable medicinal tree. New Forests, 37: 53–62.CrossRefGoogle Scholar
  43. Sim GE, Goh CJ, Loh CS. 1989. Micropropagation of Citrus mitis Blancomultiple bud formation from shoot and root explants in the presence of 6-benzylaminopurine. Plant Science, 59: 203–210.CrossRefGoogle Scholar
  44. Tiwari SK, Kashyap MK, Ujjaini MM, Agarwal AP. 2002. In vitro propagation of Lagerstromia parveflora Roxb. from adult tree. Indian Journal of Experimental Biology, 40: 212–215.PubMedGoogle Scholar
  45. Troup RS. 1986. The silviculture of Indian trees. Dilleniaceae to Leguminosae (Papilionaceae). Dehradun: International Book Distributors, Vol.2: 467–468.Google Scholar
  46. Ueda M, Tokunaga T, Okazaki M, Sata NU, Ueda K, Yamamura S. 2003. Albiziahexoside: a potential source of bioactive saponin from the leaves of Albizzia lebbeck. Natural Product Research, 17(5): 329–335.CrossRefPubMedGoogle Scholar
  47. Van Winkle SC, Johnson S, Pullman GS. 2003. The impact of gelrite and activated carbon on the elemental composition of two conifer embryogenic tissue initiation media. Plant Cell Reports, 21: 1175–1182.CrossRefPubMedGoogle Scholar
  48. Vila S, Ganzalez A, Rey H, Mroginski L. 2005. Plant regeneration, origin, and development of shoot buds from root segments of Melia azedarach L. (Miliaceae) seedlings. In Vitro Cellular and Developmental Biology-Plant, 41: 746–751.CrossRefGoogle Scholar
  49. Vinocur B, Tzfira T, Ziv M, Vainstein A, Altman A. 1997. Bud regeneration and growth from transgenic and non-transgenic aspen (Populus tremula) root explants. In: Altman A, Waisel Y (eds), Biology of Root Formation and Development. New York: Plenum Press, 217–219.Google Scholar
  50. Wang Huimei, Liu Hongmei, Wang Wenjie, Zu Yuangang. 2008. Effect of Thiadiazuron, basal medium and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus alba × P.berolinensis. Journal of Forestry Research, 19(3): 257–259.CrossRefGoogle Scholar

Copyright information

© Northeast Forestry University and Springer-Verlag Berlin Heidelberg 2011

Authors and Affiliations

  • Shahnaz Perveen
    • 1
  • Ankita Varshney
    • 1
  • Mohammad Anis
    • 1
    • 2
    Email author
  • Ibrahim M. Aref
    • 2
  1. 1.Plant Biotechnology Laboratory, Department of BotanyAligarh Muslim UniversityAligarhIndia
  2. 2.Department of Plant Production, College of Food & Agricultural SciencesKing Saud UniversityRiyadhSaudi Arabia

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