Micropropagation of Ocimum kilimandscharicum Guerke (Labiatae)

Open access

Micropropagation of Ocimum kilimandscharicum Guerke (Labiatae)

An efficient plant regeneration protocol has been developed from nodal explants of Ocimum kilimandscharicum Guerke, a medicinally important herbaceous plant species belonging to the family Lamiaceae. Axillary shoot bud proliferation was initiated from nodal explants cultured on MS medium supplemented with various concentrations of 6-benzyladenine (BA) (0.5-3.0 mg/l), kinetin (KN) (0.5-3.0 mg/l) and 2-isoPentenyladenine (2-iP) (0.5-3.0 mg/l). The maximum number of shoots (6.09±0.05), with average length 3.83±0.11 cm, was achieved with medium containing 1.0 mg/l BA. Shoot culture was established by repeated subculturing of the original nodal explants on shoot multiplication medium after each harvest of newly formed shoots. In this way, 20-30 shoots were obtained from a single nodal explant after 5 months. Rooting of shoots was achieved on half-strength MS medium supplemented with 1.5 mg/1 Indole-3-butyric acid (IBA) and 2% sucrose. Well-developed plantlets transferred to plastic pots containing soil and vermiculite (1:1) showed 81.13% survival. The genetic fidelity of in-vitro-raised field-grown plants to the donor plant was ascertained from random amplified polymorphic DNA (RAPD) markers. This protocol can be used for commercial propagation and for future genetic improvement studies.

Ahuja A, Verma M, and Grewal S. 1982. Clonal propagation of Ocimum species by tissue culture. Indian Journal of Experimental Biology 20: 455-458.

Ajitkumar D, and Seeni S. 1998. Rapid clonal multiplication through in vitro axillary shoot proliferation of Aegle marmelos (L.) Corr., a medicinal tree. Plant Cell Reports 17: 422-426.

Andrade LB, Echeverrigaray S, Fracaro F, Pauletti GF, and Rota L. 1999. The effect of growth regulators on shoot propagation and rooting of common lavender (Lavandula vera DC). Plant Cell Tissue and Organ Culture 56 (2): 79-83.

Angers P, Morales MR, and Simon JE. 1996. Basil seed oils. In: Janick J [ed.], Progress in New Crops, vol 1, 598-601. ASHS Press, Arlington, U. S. A.

Arora R, and Bhojwani SS. 1989. In vitro propagation and low temperature storage of Saussurea lappa C. B. Clarke - an endangered, medicinal plant. Plant Cell Reports 8: 44-47.

Baerts M, and Lehmann J. 1991. Veterinary medicinal plants of the region of Cretes Zaire-Nil in Burundi. Annalen Economic Wetenscbappen, Koninklijk Museum voor Midden, Africa 21: 133.

Bajaj YPS, Furmanowa M, and Olszowska O. 1988. Biotechnology of the micropropagation of medicinal and aromatic plants. In: Bajaj YPS [ed.], Biotechnology in Agriculture and Forestry, vol 24, 60-103. Medicinal and aromatic plants I. Springer-Verlag, Berlin, Germany.

Balachandran SM, Bhat SR, and Chandel KPS. 1990. In vitro clonal multiplication of turmeric (Curcuma spp.) and ginger (Zingiber officinale Rosc.). Plant Cell Reports 8: 521-524.

Banerjee S, Zehra M, and Kumar S. 1999. In vitro multiplication of Centella asiatica, a medicinal herb from leaf explants. Current Science 76: 147-148.

Banu LA, and Bari MA. 2007. Protocol establishment for multiplication and regeneration of Ocimum sanctum Linn. An important medicinal plant with high religious value in Bangladesh. Journal of Plant Science 2: 530-537.

Begum F, Amin MN, and Azad MAK. 2000. In vitro clonal propagation of Holy Basil - Ocimum sanctum L. Plant Tissue Culture and Biotechnology 10: 31-37.

Begum F, Amin MN, and Azad MAK. 2002. In vitro rapid clonal propagation of Ocimum basilicum L. Plant Tissue Culture and Biotechnology 12: 27-35.

Bekele AJ, Obeng-Ofori D, and Hassanali A. 1996. Evaluation of Ocimum suave (Wild) as a source of repellents, toxicants and protectants in storage against three stored product insect pests. International Journal of Pest Management 42: 139-142.

Bhat SR, Chandel KPS, and Malik SK. 1995. Plant regeneration from various explants of cultivated Piper species. Plant Cell Reports 14: 398-402.

Bhattacharya S, Dey T, Bandopadhyay TK, and Ghosh PD. 2008. Genetic polymorphism analysis of somatic embryo-derived plantlets of Cymbopogon flexuosus through RAPD assay. Plant Biotechnology Reports 2: 245-252.

Bhuyan AK, Pattniak SK, and Chand PK. 1997. Micropropagation of curry leaf tree (Murraya koenigii (L.) Spreng.) by axillary proliferation using intact seedlings. Plant Cell Reports 16: 779-782.

Chaudhuri RK, Pal A, and Jha TB. 2008. Conservation of Swertia chirata through direct shoot multiplication from leaf explants. Plant Biotechnology Reports 2: 213-218.

Chawdhury MKU, and Vasil JK. 1993. Molecular analysis of plants regenerated from embryogenic cultures of hybrid sugarcane cultivars (Saccharum spp.). Theoretical and Applied Genetics 86: 181-188.

Cunningham AB. 1993. African medicinal plants: setting priorities at the interface between conservation and primary healthcare. People and plants Working paper 1, 92. UNESCO, Paris.

Das P, and Pal A. 2005. Clonal propagation and production of genetically uniform regenerates from axillary meristems of adult bamboo. Journal of Plant Biochemistry and Biotechnology 14: 185-188.

De Silva T. 1997. Industrial utilization of medicinal plants in developing countries. In: Bodeder G, Bhat KKS, Burley J, and Vantomme P [eds.], Medicinal Plants Forest Conservation and Healthcare. Non-Wood Forest Products No.11. FAO, Rome, Italy.

Debergh PC, and Maene LJ. 1981. A scheme for commercial propagation of ornamental plants by tissue culture. Scientia Horticulturae 14: 335-345.

Demissew S, and Asfaw N. 1994. Some useful indigenous labiates from Ethiopia. Lamiales Newsletter 3: 5-6.

Dewir YH, Chakrabarty D, Hahn EJ, and Paek KY. 2005. Reversion of inflorescence development in Euphorbia millii and its application to large-scale micropropagation in an air-lift bioreactor. Journal of Horticultural Science and Biotechnology 80: 581-587.

Dode LB, Bobrowski VL, Braga EJB, Seixas FK, and Schuch MW. 2003. In vitro propagation of Ocimum basilicum L. Maringa 25: 435-437.

Driver JA, and Suttle GR. 1987. Nursery handling of propagules. In: Bonga JM, and Durzan DJ [eds.], Cell and Tissue Culture in Forestry, 320-335. Dordrecht, Netherlands.

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

Faisal M, and Anis N. 2003. Rapid mass propagation of Tylophora indica Merrill via leaf callus culture. Plant Cell Tissue and Organ Culture 75: 125-129.

Flower MW. 1983. Commercial application and economic aspects of mass cell cultures. In: Mantel SH, and Smith H [eds.], Plant Biotechnology, 3-37. Cambridge University Press, Cambridge-UK.

George EF. 1993. Plant Propagation by Tissue Culture. Part-I. The Technology Exegetics Ltd, Edington.

George PS, Ravishankar GA, and Venkataraman LV. 1993. Clonal multiplication of Gardenia jaminoides Elllis through axillary bud culture. Plant Cell Reports 13: 59-62.

Gopi C, Sekhar Nataraja Y, and Ponmurugan P. 2006. In vitro multiplication of Ocimum gratissimum L., through direct regeneration. African Journal of Biotechnology 5: 723-726.

Hakkim FL, Arivazhagan G, and Boopathy R. 2008. Antioxidant property of selected Ocimum species and their secondary metabolite content. Journal of Medicinal Plants Research 2: 250-257.

Hassanali A, Lwande W, Ole-Sitayo N, Morkeka L, Nokoe S, and Chapya A. 1990. Weevil repellent constituents of Ocimum suave leaves and Eugenia caryophyllata cloves used as grain protectant in parts of Eastern Africa. Discovery and Innovations 2: 91-95.

Heywood VH. 1978. Flowering Plants of the World, 239-240. Oxford Univ. Press, UK.

Hiregoudar LV, Murthy HN, Bhat JG, Nayeem A, Hema BP, Hahn EJ, and Paek KY. 2006. Rapid clonal propagation of Vitex trifolia.Biologia Plantarum 50: 291-294.

Hiregoudar LV, Murthy HN, Hema BP, Hahn EJ, and Paek KY. 2003. Multiple shoot induction and plant regeneration of Feronia limonia (L.) Swingle. Scientia Horticulturae 98: 357-364.

Hossain PJ, Biswas BK, Karim MR, Rahman S, Islam R, and Jorder OI. 1994. In vitro organogenesis of elephant apple (Feronia limonia). Plant Cell Tissue and Organ Culture 39: 265-268.

Jembere B, Hassanali A, and Obeng-Ofori D. 1994. Scientific rational of traditional post-harvest insect control practices: lessons from studies on Ocimum species. Proceedings 3rd International Conference on Tropical Entomology. 30th October-4th November, Nairobi, Kenya.

Jembere B, Obeng-Ofori D, Hassanali A, and Nyamasyo GHN. 1995. Products derived from the leaves of Ocimum kilimandscharicum (Labiatae) as post harvest protectant against the infestation of three major stored product insect pests. Bulletin Entomological Research 85: 361-367.

Kokwaro JO. 1976. Medicinal Plants of East Africa, 384. East African Literature Bureau, General Printers Limited, Nairobi.

Krishnan PN, and Seeni S. 1994. Rapid micropropagation of Woodfordia fruticosa (L.) Kurz. (Lythraccae), a rare medicinal plant. Plant Cell Reports 14: 55-58.

Kukreja AK, Mathur AK, and Zaim M. 1990. Mass production of virus free patchouli plants [Pogostemon cabin (Blanco) Benth] by in vitro culture. Tropical Agriculture 67: 101-104.

Kweka EJ, Mosha FW, Lowassa A, Mahande AM, Mahande MJ, Massenga CP, Tenu F, Lyatuu EE, Mboya MA, and Temu EA. 2008. Longitudinal evaluation of Ocimum and other plants effect on the feeding behavioral response of mosquitoes (Diptera: Culicidae) in the field in Tanzania. Parasites and Vectors 1: 42.

Lakshmanan KK, and Dhanalakshmi S. 1990. Callus, organogenesis and plantlet formation in tissue cultures of Clitoria ternatea.Annals of Botany 66: 451-455.

Mauseth JD. 1979. A new method for the propagation of cacti: sterile culture of axillary buds. Cactus and Succulent Journal 51: 186-187.

Morales MR, Charles DJ, Simon JE. 1993b. New aromatic lemon basil germplasm. In: Janick J, and Simon JE [eds.], New Crops, 632-635. John Wiley and Sons, New York, U. S. A.

Murashige T, and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15: 473-497.

Murray MG, and Thompson WF. 1980. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Research 8: 4321-4325.

Obeng-Ofori D, and Reichmuth C. 1997. Biological activity of eugenol, a major component of Ocimum suave (Wild) against four species of stored product coleopteran. International Journal of Pest Management 43: 89-94.

Obeng-Ofori D, Reichmuth CH, Bekele AJ, and Hassanali A. 1998. Toxicity and protectant potential of camphor, a major component of essential oil of Ocimum kilimandscharicum, against four stored product beetles. International Journal of Pest Management 44: 203-209.

Patnaik J, and Debata BK. 1996. Micropropagation of Hemidesmus indicus (L.) R. Br. through axillary bud culture. Plant Cell Reports 15: 473-497.

Paton A. 1991. A synopsis of Ocimum (Labiatae) in Africa. Kew Bulletin 47: 403-435.

Prakash E, Sha Valli Khan PS, Sairam Reddy P, and Rao KR. 1999. Regeneration of plants from seed-derived callus of Hybanthus enneaspermus L. Muell., a rare ethnobotanical herb. Plant Cell Reports 18: 873-878.

Prasad G, Kumar A, Singh AK, Bhattacharya AK, Singh K, and Harma VD. 1986. Antimicrobial activity of essential oils of some Ocimum species and clove oil. Fotpterapia 57: 429-432.

Purohit SD, and Tak K. 1992. In vitro propagation of an adult tree Feronia limonia L. through axillay branching. Indian Journal of Experimental Biology 30: 377-399.

Purohit SD, Dave A, and Kukda G. 1994. Micropropagation of safed musli (Chlorophytum borivilianum), a rare Indian medicinal plant. Plant Cell Tissue and Organ Culture 39: 93-96.

Rani V, Parida A, and Raina SN. 1995. Random amplified polymorphic DNA (RAPD) markers for genetic analysis of micropropagated plants of Populus deltoids Marsh. Plant Cell Reports 14: 459-462.

Rout GR, DAS P, Goel S, and Raina SN. 1998. Determination of genetic stability of micropropagated ginger plants using Random Amplified Polymorphic DNA (RAPD) markers. Botanical Bulletin of Academia Sinica 39: 23-27.

Saha S, Ghosh PD and Sengupta C. 2010. An efficient method for micropropagation of Ocimum basilicum L. Indian Journal of Plant Physiology 15 (2): 168-172.

Saha S, Ghosh PD and Sengupta C. 2010. In vitro multiple shoot regeneration of Mentha piperita.Journal of Tropical Medicinal Plants 11(1): 89-92.

Sahoo Y, and Chand PK. 1998. Micropropagation of Vitex negundo L., a woody aromatic medicinal shrub, through high-frequency axillary shoot proliferation. Plant Cell Reports 18: 301-307.

Sahoo Y, Pattnaik SK, and Chand PK. 1997. In vitro clonal propagation of an aromatic medicinal herb Ocimum basilicum L. (sweet basil) by axillary shoot proliferation. In Vitro Cellular and Developmental Biology - Plant 33: 293-296.

Sambrook J, and Russed DW. 2001. Molecular Cloning. A Laboratory Manual 3rd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, YN.

Saxena S, and Bhojwani SS. 1993. In vitro clonal multiplication of 4-year-old plants of the bamboo Dendrocalamus longispathus Kurz. In Vitro Cellular and Developmental Biology - Plant 29(3): 135-142.

Sen J, and Sharma AK. 1991. Micropropagation of Withania somnifera from germinating seeds and shoot tips. Plant Cell Tissue and Organ Culture 26: 71-73.

Shetty K. 1997. Biotechnology to harness the benefits of dietary phenols: focus on Lamiaceae. Asia Pacific Journal of Clinical Nutrition 6: 162-171.

Siddique I, and Anis M. 2007a. In vitro shoot multiplication and plantlet regeneration from nodal explants of Cassia angustifolia (Vahl): a medicinal plant. Acta Physiolgiae Plantarum 29: 233-238.

Simon JE, and Reiss-Bubenheim D. 1987. Characteristics of American basil varieties. In: Simon JE, and Grant L [eds.], Proceedings of the Second National Herb Growing and Marketing Conference, 48-51. Indianapolis, Indiana, U. S. A.

Singh NK, and Sehgal CB. 1999. Micropropagation of ‘Holy Basil’ (Ocimum sanctum Linn.) from young inflorescences of mature plants. Plant Growth Regulation 29: 161-166.

Sudha GC, and Seeni S. 1994. In vitro propagation and field establishment of Adhatoda beddomei C. B. Clarke, a rare medicinal plant. Plant Cell Reports 13: 203-207.

Tiwari KN, Sharma NC, Tiwari V, and Singh BD. 2000. Micropropagation of Centella asiatica (L.), a valuable medicinal herb. Plant Cell Tissue and Organ Culture 63: 179-185.

Tiwari V, Tiwari KN, and Singh BD. 2001. Comparative studies of cytokinins on in vitro propagation of Bacopa monniera. Plant Cell Tissue and Organ Culture 66: 9-16.

Viera RE, Fahoer R, and Raina SK. 2001. Genetic Diversity of Ocimum gratissimum L. based on volatile oil constituents, flavonoids and RAPD markers. Biochemical Systematics and Ecology 29: 287-304.

Vincent KA, Mary Mathew K, and Hariharan M. 1992. Micropropagation of Kaemferia galanga L. - a medicinal plant. Plant Cell Tissue and Organ Culture 28: 229-230.

Acta Biologica Cracoviensia s. Botanica

The Journal of Polish Academy of Sciences

Journal Information

IMPACT FACTOR 2016: 0.491
5-year IMPACT FACTOR: 0.787

CiteScore 2016: 0.51

SCImago Journal Rank (SJR) 2016: 0.242
Source Normalized Impact per Paper (SNIP) 2016: 0.264

Ministry of Science and Higher Education: 20 points

Cited By


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 193 193 27
PDF Downloads 193 193 36