Summary
A rapid micropropagation system was established forHolostemma annulare (Roxb.) K. Schum., (H. ada-kodien R. Br. ex Schult; Asclepiadaceae), a rare medicinal plant. Shoot tips (0.5–0.8 cm) and terminal and basal nodes (1.0–1.5 cm) harvested from actively growing shoots of conventionally raised plants were cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzyladenine (BA) and α-naphthaleneacetic acid (NAA). Multiple shoot formation (3.8) was observed in 68% of basal nodes cultured on medium with optimum concentration of 4.43 μM BA and 0.54 μM NAA after 8 wk. Terminal nodes were not suitable for inducing multiple shoots. Irrespective of the orientation (vertical/horizontal), all shoot tip explants responded with a single shoot in all the combinations of plant growth regulators tried. Effects of other cytokinins (kinetin and 2-isopentenyladenine) and auxins [indole-3-acetic acid and indole-3-butyric acid (IBA)] to enhance the regeneration potential of basal nodes were analyzed. Shoots were multiplied by subculture of basal nodes and stumps (the original explant tissue free of shoots, but with remnant axillary, meristem and two or three protruding buds) in a reduced concentration of BA (2.21 μM) and NAA (0.27 μM). Liquid medium for multiplication was found to be ineffective due to a high degree of hyperhydricity. To make the multiplication process cost effective, culture bottles with polypropylene, caps were used for multiplication. The best root induction (75%) and survival (80%) was achieved on 0.5 strength MS medium supplemented with 1.48 μM IBA. Field-established plants had uniform growth habit traits in terms of height of plants and number, length, and weight of the tuberous roots.
Similar content being viewed by others
References
Anonymous. The Wealth of India: raw materials. Vol. V. Council of Scientific and Industrial Research New Delhi; 1959:111.
Bajaj, Y. P. S.; Furmanowa, M.; Olszowskao, O. Biotechnology of the micropropagation of medicinal and aromatic plants. In: Bajaj, Y. P. S., ed. Biotechnology in agriculture and forestry. Vol. 4. New York: Springer-Verlag; 1988:60–103.
Bateson, J. M.; Grout, B. W. W.; Lane, S. The influence of container dimensions on the multiplication rate of regenerating plant cell cultures. Symp. Florizel, Plant Micropropagation in Horticultural Industries, Belgium; 1987.
CAMP-I, The first red list of medicinal plants of South India. Foundation for Revitalization of Local Health Tradition (FRLHT), Anandnagar, Bangalore, India; 1995.
Dan, M.; Shanavaskhan, A. E. A glance to some rare medicinal plants of Western Ghats. In: Karunakaran, K. C., ed. Proceedings of the Symposium on Rare, Endangered, and Endemic Plants of the Western Ghats. Kerala Forest Department, Trivandrum; 1991:221–226.
Debergh, P. C. Effect of agar brand and concentration on the tissue culture medium. Physiol. Plant. 59:270–276; 1983.
Ebert, A.; Taylor, F.; Blake, J. Changes of 6-benzylaminopurine and 2,4-dichlorophenoxyacetic acid concentrations in plant tissue culture media in the presence of activated charcoal. Plant Cell Tissue Organ Cult. 33:157–162; 1993.
Gulati, A.; Jaiswal, P. K. In vitro induction of multiple shoots and plant regeneration from shoot tips of mung bean (Vigna radiata (L.) Wilczek). Plant Cell Tissue Organ Cult. 29:199–205; 1992.
Hossain, M.; Karim, M. R.; Islam, R., et al. Plant regeneration from nucellar tissues ofAegle marmelos through organogenesis. Plant Cell Tissue Organ Cult. 34:199–203; 1993.
Huber, H. Asclepiadaceae. In: Dassamayate, M. D.; Fosbery, F. R., ed. A resived handbook to the flora of Ceylon. 4:81; 1983.
Kolammal, M., ed. Pharmacognosy of Ayurvedic drugs. Kerala Ser. 10, Dept. Pharmacognosy, University of Kerala Trivandrum; 1979:21.
Krishnan, P. N.; Sudha, C. G.; Seeni, S. Rapid propagation through shoot tip culture ofTrichopus zeylanicus Gaertn., a rare ethnomedicinal plant. Plant Cell Rep. 14:708–711; 1995.
Mao, A. A.; Wetten, A.; Caligari, P. D. S. In vitro propagation ofClerodendrum colebrookianum Walp., a potential natural antihypertensive medicinal plant. Plant Cell Rep. 14:493–496; 1995.
Matthew, K. M. The flora of the Tamil Nadu-Carnatic Part II. The Rapinat Herbarium, Thiruchirapally, India. 1983:944–50.
McCown, B. H.; Sellmer, J. C. General media and vessels suitable for woody plant culture. In: Bonga, J. M.; Durzan, D. J., ed. Cell and tissue culture in forestry. Vol. 1. The Netherlands: Martinus Nijhoff, Publisher; 1987:4–16.
Murashige, T.; Skoog, F. A revised medium, for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–497; 1962.
Nair, K. V.; Nair, A. R.; Nair, C. P. R. Technoeconomic data on cultivation, preservation of some south Indian medicinal plants. Aryavaidyan 5:238–240; 1992.
Norton, M. E.; Norton, C. R. Explant origin as a determinant ofin vitro shoot proliferation inPrunus andSpiraea. J. Hort. Sci. 61:43–48; 1986.
Okubo, H.; Wada, K.; Uemoto, S.In vitro morphogenic response and distribution of endogenous plant hormones in hypocotyl segments of snapdragon (Antirrhinum majus L.). Plant Cell Rep. 10:501–504; 1991.
Pereira, A. M. S.; Moro, J. R.; Cerdeira, R. M. M., et al. Effect of phytoregulators and physiological characteristics of the explants on micropropagation ofMaytenus ilicifolia. Plant Cell Tissue Organ Cult. 42:295–297; 1995.
Phan, C.T. Vitreous state inin vitro culture: ethylene versus cytokinin. Plant Cell Rep. 9:517–519; 1991.
Pramanik, T. K.; Datta, S. K. Plant regeneration and ploidy variation in culture derived plants ofAsclepias currassavica L. Plant Cell Rep. 3:219–222; 1986.
Purohit, S. D.; Dava, A.; Kukda, G. Micropropagation of safed musli (Chlorophytum borivilianum), a rare Indian medicinal herb. Plant Cell Tissue Organ Cult. 39:93–96; 1994.
Quraishi, A.; Koche, V.; Mishra, S. K. In vitro micropropagation from nodal segments ofCleistanthus collinus. Plant Cell Tissue Organ Cult. 45:87–91; 1996.
Raghavaswamy, B. V.; Himabindu, K.; Lakshmi, S. G.,In vitro micropropagation of elite rosewood (Dalbergia latifolia Roxb). Plant Cell Rep. 11:126–131; 1992.
Ripley, K. P.; Preece, J. E. Micropropagation ofEuphorbia lathyris L. Plant Cell Tissue Organ Cult. 5:213–218; 1986.
Roy, A. T.; De, D. N. Tissue culture and plant regeneration from immature embryo explants ofCalotropis gigantea (Linn.) R. Br. Plant Cell Tissue Organ Cult. 20:229–233; 1990.
Sallanon, H.; Maziere, Y. Influence of growth room and vessel humidity on thein vitro development of rose plants. Plant Cell Tissue Organ Cult. 30:121–125; 1992.
Sharma, N.; Chandal, K. P. S. Effect of ascorbic acid on axillary shoot induction inTylophora indica (Burm. f.) Merrill. Plant Cell Tissue Organ Cult. 29:109–113; 1992a.
Sharma, N.; Chandal, K. P. S. Low temperature storage ofRauvolfia serpentina Benth. ex Kurz., an, endangered endemic medicinal plant Plant Cell Rep. 11:200–203; 1992b.
Sudha, C. C.; Seeni, S. In vitro propagation ofRauwolfia micrantha, a rare medicinal plant. Plant Cell Tissue Organ Cult. 44:243–248; 1996.
Weatherhead, M. A.; Burdon, J.; Henshaw, G. G. Effect of activated charcoal as an additive to plant tissue culture media. Part 2. Z. Planzenphysiol 94:399–406; 1979.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sudha, C.G., Krishnan, P.N. & Pushpangadan, P. In vitro propagation ofHolostemma annulare (Roxb.) K. Schum., a rare medicinal plant. In Vitro Cell.Dev.Biol.-Plant 34, 57–63 (1998). https://doi.org/10.1007/BF02823124
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02823124