In vitro callus and shoot regeneration in Enterolobium cyclocarpum (Jacq.) Grised. - a fast timber yielding species


  • Michael S. AKINROPO Obafemi Awolowo University, Department of Botany, Ile-Ife, Osun State (NG)
  • Benjamin E. AYISIRE Obafemi Awolowo University, Department of Botany, Ile-Ife, Osun State (NG)
  • Ejeoghene R. OGBIMI Obafemi Awolowo University, Department of Botany, Ile-Ife, Osun State (NG)



callus; caulogenesis; callogenesis; explants; multiple shoots


This study was conducted to investigate the in vitro callus induction and rapid shoot regeneration potential in Enterolobium cyclocarpum, a plant native to central Mexico but widely introduced into Africa. The leaf, stem and nodal explants of E. cyclocarpum were cultured on full strength Murashige and Skoog (MS) medium supplemented with different concentrations of Cytokinins - Benzyladenine (BA) and/or Kinetin and Auxins - Naphthalene acetic acid (NAA) and/or 2,4-Dichlorophenoxylacetic acid (2,4-D) each alone and in combination.  The leaf explants did not respond to these treatments.  The Nodal explants were best for caulogenesis, while the explant responses were in the order- nodal > stem > cotyledon for callogenesis in MS medium supplemented with BA and/or Kin combined with NAA and/or 2,4-D. The varied combinations induced white compact callus.  The highest callus production was observed on MS medium supplemented with 2.7 μM NAA + 2.2 μM BA and 5.4 μM NAA alone.  Nodal and cotyledon explants developed callus and multiple shoots on MS supplemented with a combination of cytokinin (BA and/or Kin.) and auxin (NAA and/or 2,4-D). The maximum number of 3.98 ± 0.37 and 2.1±0.11 shoots/explants were recorded for nodal and cotyledon explants on MS medium supplemented with a combination of 8.8 μM BA+2.7 μM NAA and 2.2μM BA+2.7 μM NAA respectively.  On the basal medium, 10% of the excised shoots rooted successfully. Thus, this in vitro method can be exploited for conservation and mass propagation of this fast timber yielding tree and also utilized for embryogenesis studies.


Metrics Loading ...


Abbasin Z, Zamani S, Movahedi S, Khaksar G, Tabatabael (2010). In vitro micropropagation of yew (Taxus baccata) and propagation of plantlets. Biotechnology 9:48-54.

Agrawal V, Sardar PR (2007). In vitro propagation of Cassia angustifolia through leaflet and cotyledon-derived calli. Biologia plantarum 50(1):118-122.

Anis M, Husain MK, Shahzad A (2005). In vitro plantlet regeneration of Pterocarpus marsupium Roxb., an endangered leguminous tree. Current Science 88:861- 863.

Araceli RS, Osvaldo ACH, Gustavo JAH (2007). In vitro propagation of Enterolobium cyclocarpum (Guanacaste) from nodal explants of axenic seedlings e-Gnosis. Vol. 5, Universidad de Guadalajara, Guadalajara, Mexico.

Bairu MW, Kane ME (2011). Physiological and developmental problems encountered by in vitro cultured plants. Plant Growth Regulation 63:101-103.

Banerjee M, Gantait S, Pramanik BR (2011). A two-step method for accelerated mass propagation of Dendrocalamus asper and their evaluation in field. Physiology and Molecular Biology of Plants 17(4):387-393.

Barwick M (2004). Tropical and subtropical trees - a worldwide encyclopedic guide. ISBN 0-500-51181-0P.

Burkil HM (2004). The useful plants of west tropical Africa. Royal Botanic Gardens, Kew.

Carranza-Montaño MA, Sánchez-Velásquez LR, Pineda-López MR, Cuevas-Guzmán R (2003). Forrage quality and potential of species from the Sierra de Manantlán (México) tropical dry forest. Agrociencia 37(2):203-210.

Chalupa V (1981) Clonal Propagation of broad-leaved forest trees in vitro. Communications Institute Forestry Czechoslovakia 12:255-271.

Chalupa V (2002) In vitro propagation of mature trees of Sorbus aucuparia L. and field performance of micropropagated trees. Journal of forest science 48(12):529-535.

Chen Y, Cao FL, Li SX, Dao DW, Xu CP (2010). Establishment of highly efficient regeneration system with different explants of Sapium sabiferum in vitro. Acta Botanica Boreali-Occidentalia Sinica 30(12):2542-2549 (in Chinese).

Chevre, AM (1985). Research on in-vitro vegetative propagation in chestnut. Doctoral Thesis. University of Bordeaux II, pp 100.

de Oliveira Y, Pinto F, da Silwa ALL, Guedas I, Biasi LA, Quoirin M (2010). An efficient protocol for micropropagation of Melaleuca alternifolia Cheel. In Vitro Cell Developmental Biology - Plant 46:192-200.

Dhar U, Joshi M (2005). Efficient plant regeneration protocol through callus for Saussurea obvalata (Asteraceae): effect of explant type, age and plant growth regulators. Plant Cell Reports 24:195-200.

Eleanya K (2014). Promoting forests and forest products development. Production Agriculture and Technology 10(2):78-90.

Ezenwa I (1999). Preliminary evaluation of the suitability of Enterolobium cyclocarpum for use in intensive feed garden in Southwestern Nigeria. Agroforestry System 44(1):13-19.

Faisal M, Ahmad N, Anis M (2007). An efficient micropropagation system for Tylophora indica: an endangered medicinally important plant. Plant Biotechnology Reports 1:155-161.

Food and Agricultural Organization FAO (2005) Forest Resources Assessment: FAO Rome.

Farinola LA, Famuyide OO, Adebayo O, Awe F, Adedokun FT (2013). Overview of the link between green economy and sustainable rural livelihood in Nigeria. FAN Conference, pp 659-668.

FORMECU (1991). Forestry Plantation Development in Nigeria by 1990 Formecu/stat/publication no. 14, Ibadan, Nigeria.

Gostin I (2008). Effects of different plant hormones on Salvia officinalis cultivated in vitro. International Journal of Botany 4:430-436 .

Handro W, Floh EJ (2001). Neo-formation of flower buds and other morphorgenetic responses in tissue culture of Melia azedarach. Plant Cell, Tissue and Organ Culture 64:73-76.

Husain MK, Ani M, Shahzad A (2008). In vitro propagation of multipurpose leguminous tree Pterocarpus marsupium Roxb. using nodal explants. Acta Physiologiae 30:353-35.

Isikalan C, Akbas F, Namli S, Basaran D (2010). Adventitious shoot development from leaf and stem explants of Amygdalus communis L. cv. ‘Yaltunski’. Plant Omics Journal 3(3):92-96.

Jain SM, De Klerk GJ (1998). Somaclonal variation in breeding and propagation of ornamental crops. Plant Tissue Culture and Biotechnology 4:63-75.

Jiménez VM (2001). Regulation of in vitro somatic embryogenesis with emphasis on the role of endogenous hormones. Revista Brasileira de Fisiologia Vegetal 13:196-223.

Kahl G (1983). Wound repair and tumor induction in higher plants. In: Akazawa T, Imasei H (Eds). The new frontiers in plant biochemistry. Japan Scientific Society Press/ Martinus Nijhoff/ Dr. Junk W Press, Tokyo and the Hague, pp 193- 216.

Ladipo D (2010). The state of Nigeria’s forest. International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria.>issue4/

Lin J, Wu L, Liang J, Wang J (2010). Effect of different plant growth regulators on callus induction in Catalpa bungei. African Journal of Agricultural Research 5(19):2699-2704.

Malik SK, Kalia RK, Chaudhury R (2010). In vitro regeneration of Garcinia indica using leaf explants. Indian Journal of Plant Physiology 15:262-266.

Martin KP (2002). Rapid Propagation of Holostemma adakodien Schult., a rare medicinal plant through axillary bud multiplication and indirect organogenesis. Plant Cell Reports 21:112-117.

Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15:473-495.

Naval MM, Llacer G, Badenes ML, Glordani E (2009). Adventitious shoot regeneration from leaf explants of the persimmon (Diospyras kaki Thumb.) cv ‘Rojo Brillante’. Acta Horticulturae 833:183-186.

Niembro-Rocas A (2003). Enterolobium cyclocarpum (Jacq.) Griseb. In: Vozzo JA (Ed). Tropical tree seed manual. US Department of Agriculture Handbook 721. USDA Forest Service, Washington, DC.

Ntui VO, Uyoh EA, Urua IS, Ogbu U, Okpako EC (2012). Regeneration of Parkia biglobosa Benth.: an important tree species of Africa. Journal of Microbiology and Biotechnology Research 2(1):169-177.

Odediran FA, Arabomen O, Akanbi FS, Obafunsho OE, Wahab WT (2013). Deforestation: causes and consequences on the environment and resource sustainability in Nigeria. Proceedings of the FAN conference, pp 484-492.

Pérez-Francés JF, Valdés F, Msrtín R (1995) Callus induction and culture from explants of Erysimum scopariumin a growth regulator-free medium. Plant Cell, Tissue and Organ Culture 43(3):223-228.

Rahman SM, Hossain M, Biswas BK, Joarder OI, Islam R (1993). Micropropagation of Caeselpina pucherima through nodal bud culture of mature tree. Plant Cell, Tissue and Organ Culture 32:363-365.

Razdan MK (2003). Organogenesis. In: Introduction to plant tissue culture. 2nd Ed. Science Publishers Inc. Enfield (NH), USA, Plymouth UK pp 71.

Renukdas N, Manoharan M, Garner JO (2010). In vitro propagation of pecan (Carya illinoinensis (Wangenh) R. Koch). Plant Biotechnology 27:211- 215.

Rincón F, Oberto A, León-De Pinto G (2005). Functionality of Enterolobium cyclocarpum gum in the preparation of liquid low-fat yogurt. Revista Cientifica-Faculdad de Ciencias Veterinarias 15(1):83-87.

Rincón F, León-De Pinto G, Beltrón O (2006). Behaviour of a mixture of Acacia glomerosa, Enterolobium cyclocarpum and Hymenaea courbaryl gums in ice cream preparation: note. Food Science and Technology International 12(1):13-17.

Rita I, Floh, EIS (1995). Tissue culture and micropropagation of Cuphea ericoides, a potential source of medium-chain fatty acids. Plant Cell, Tissue and Organ Culture 40:187-189.

Rocha SC, Quoirin M (2004). Calogênese e rizogênese em explants de mogno (Swietenia macrophylla King) cultivados in vitro. Ciência Florestal 14:91-101.

Sadegh M, Rosna MT, Arash K, Jamilah SY (2012). Induction of different types of callus and somatic embryogenesis in various explants of Sainfoin (Onobrychis sativa). Australian Journal of Crop Science 6(8):1305-1313.

Sajeevan R Singh SJ, Nataraj KN, Shivanna MB (2011). An efficient in vitro protocol for multiple shoot induction in Mulberry, Morus alba L. variety VI. International Research Journal for Plant Science 2(8):254-261.

Salvi ND, Singh H, Tivarekar S, Eapen S (2001). Plant regeneration from different explants of Neem. Plant Cell, Tissue and Organ Culture 65:159-162.

Sayer AJ, Harcourt CS, Collins MN (2010). The conservation atlas of tropical forest Africa. Macmillan Publisher Ltd, ISBN 033337757-4, pp 9-230.

Shasthree T, Imran MA, Mallaiah B (2009). In vitro rooting from callus cultures derived from seedling explants of Erythrina variagata L. Current Trends in Biotechnology and Pharmacy 3(4):447- 452.

Sugla T, Purkayastha J, Krant, KS, Solleti SK, Sahoo L (2007). Micropropagation of Pongamia pinnata through enhanced axillary branching. In vitro Cellular and Developmental Biology in Plant 43(5):409-414.

Standley PC, Steyermark JA (1946). Enterolobium in flora of Guatemala. Fieldiana: Botany 24(5):32-34.

Thomas TD, Shankar S (2009). Multiple shoot induction and callus regeneration in Sarcostemma brevistigma Wight and Arnott, a rare medicinal plant. Plant Biotechnology Reports 3(1):67-74.

Tibok A, Blackhall NW, Power JB, Davey (1999). Optimized plant regeneration from callus derived from seedling hypocotyls of Eucalyptus urophylla. Plant Science 110(1):139-145.

Trevor AT, Indra, SSH (1990). Special problem and prospects in the propagation of woody species. In: Plant aging Springer, Boston, MA, pp 67-74.

United Nation Environmental Programme: UNEP (2003). http://www.uneporg/DEWA/Africa/docs/en/aeo

UN-REDD (2013). The State of Nigeria’s Forest. Programme/Nigeria/tabid/199/default.aspx

Uphof JC (1959). The dictionary of economic plants. Weinheim, J Cramer.

Venkatachalam P, Geetha N, Khandelwal A, Shaila MS, Sita GL (1999). Induction of direct somatic embryogenesis and plant regeneration from mature cotyledon explants of Arachis hypogaea L. Current Science 77:269-273.

Vidoz ML, Quesenberry KH, Real D, Gallo M (2012). Plant regeneration of Lotononis bainesii Baker (Fabaceae) through cotyledon and leaf culture. African Journal of Biotechnology 11(41):9724-9731.

Xie D, Hong Y (2001). In vitro regeneration of Acacia magnum via organogenesis. Plant Cell, Tissue and Organ Culture 66:167-173.

Xiaomei Liu, Pijut PM (2008). Plant regeneration from in vitro leaves of mature black cherry (Prunus serotina). Plant Cell, Tissue and Organ Culture 94:113-123.

Ye X, Chen y, Li J, Yu X, Feng J, Zheng X (2012). Callus induction and adventitious shoot regeneration in Zizyphus jujuba Mill. ‘Huizao’. African Journal of Biotechnology 11(16):3888-3894.

Yin Z, Zhiqi Z, Jianjun Z (2001). Use of hymexazol (HMI) in rapid propagation of Albizia julibrissin Durazz. Acta Agriculturae Shanghai 17:31-34.




How to Cite

AKINROPO, M. S., AYISIRE, B. E. ., & OGBIMI, E. R. . (2020). In vitro callus and shoot regeneration in Enterolobium cyclocarpum (Jacq.) Grised. - a fast timber yielding species. Notulae Scientia Biologicae, 12(1), 74–89.



Research articles
DOI: 10.15835/nsb12110604