Optimized conditions for callus induction, plant regeneration and alkaloids accumulation in stem and shoot tip explants of Phyla nodiflora

  • A.B.A. Ahmed Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia Department of Plant Science. Bharathidasan University. Tiruchirappalli-620024, Tamil Nadu, India
  • R. Pallela Marine Bioprocess Research Center, Pukyong National University, Busan-608737, South Korea
  • A.S. Rao Department of Biotechnology. Bharathidasan University. Tiruchirappalli-620024, Tamil Nadu, India
  • M.V. Rao Department of Plant Science. Bharathidasan University. Tiruchirappalli-620024, Tamil Nadu, India
  • R. Mat Taha Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia
Keywords: callus biomass, gravimetric method, growth curve, HPTLC, rooting, TLC

Abstract

The present study describes callus induction and the subsequent plant regeneration with alkaloids accumulation instem and shoot tip explants of Phyla nodiflora. Both explants were cultured on different media (MS, B5, SH and WPM) for callus induction. Stem explants showed better callus biomass (dry weight) than shoot tip explants with green compact callus when cultured on MS medium containing 1.5 mg L–1 α-naphthalene acetic acid. Shoots were regenerated from the callus on MS medium with 1.5 mg L–1 α-naphthalene acetic acid and 1.0 mg L–1 benzyl adenine. The rooting of all regenerated shoots was successfully performed on half-strength MS medium with 1.0 mg L–1 indole-3-butyric acid. The plantlets were acclimatized and established in soil (90%) and exhibited morphological characteristics similar to those of the mother plant. In addition, the alkaloids content was higher in regenerated callus than intact stem and shoot tip explants, which were analyzed by a gravimetric method, TLC (thin layer chromatography) and HPTLC (high performance thin layer chromatography). The proposed method could effectively be applied for the conservation and clonal propagation to meet the pharmaceutical demands of this medicinally important species

Downloads

Download data is not yet available.

References

Abbasi A.M., Khan M.A., Ahmad M., Zafar M., Jahan S., Sultana S., 2010. Ethnopharmacological application of medicinal plants to cure skin diseases and in folk cosmetics among the tribal communities of north-west frontier province. Pakistan J Ethnopharmacol 128, 322-335. http://dx.doi.org/10.1016/j.jep.2010.01.052

Agarwal V.S., 1997. Drug plants of India. Vol. II. Kalyani Publ., New Delhi. pp. 82-83.

Ahmed A.B.A., Gouthaman T., Rao A.S., Rao M.V., 2005. Micropropagation of Phyla nodiflora (L.) Greene: an important medicinal plant. Iranian J Biotech 3, 186-190.

Ahmed A.B.A., Rao A.S., Rao M.V., 2010. In vitro callus and in vivo leaf of Gymnema sylvestre stimulate β-cells regeneration and anti-diabetic activity in Wistar rats. Phytomedicine 17, 1033-1039. http://dx.doi.org/10.1016/j.phymed.2010.03.019

Ahmed A.B.A., Rao A.S., Rao M.V., Taha R.M., 2011. Effect of picloram, additives and plant growth regulators on somatic embryogenesis of Phyla nodiflora (L.) Greene. Braz Arch Biol Tech 54, 7-13. http://dx.doi.org/10.1590/S1516-89132011000100002

Balaraju K., Agastian P., Preetamraj J.P., Arokiyaraj S., Ignacimuthu S., 2008. Micropropagation of Vitex agnus-castus (Verbenaceae) – a valuable medicinal plant. In Vitro Cell Dev Biol–Plant 44, 436-441. http://dx.doi.org/10.1007/s11627-008-9155-9

Forestieri A.M., Monforte M.T., Ragusa, S., Trovato, A., Jauk, L., 1996. Anti-inflammatory, analgesic and antipyretic activity in Rodents of plant extracts used in African medicine. Phytother Res 10, 100-106. 3.0.CO;2-I" target="_blank">http://dx.doi.org/10.1002/(SICI)1099-1573(199603)10:2<100::AID-PTR724>3.0.CO;2-I

GAMBORG O.L., MILLER R.A., OJIMA K., 1968. Nutrients requirements of suspension culture of soybean root cells. Exp Cell Res 50, 151-158.

Georges D., Chenieux J.C., Ochatt S.J., 1993. Plant Regeneration from Aged-Callus of the Woody Ornamental Species Lonicera japonica cv. Hall's Prolific. Plant Cell Rep 13, 91-94. http://dx.doi.org/10.1007/BF00235297

Gomez K.A., Gomez A.A., 1976. Statistical procedures for agricultural research with emphasis on rice. International Rice Research Institute, Los Bans, Philippines: IRRI Publ., p 264.

Gupta S.K., Khanuja S.P.S., Kumar S., 2001. In vitro Propagation of Lippia alba. Curr Sci 81, 206-210.

Harborne J.B., 1973. Phytochemical Methods, Chapman and Hall, London, pp.11-21.

Hussain I.A., Rashid M.H., Qurashi A., 2001. In vitro multiplication of Gladiolus. Plant Tiss Cult 5, 27-33.

Joshi C.S., Priya E.S., Venkataraman S., 2007. Acute and subacute toxicity studies on the polyherbal antidiabetic formulation diakyur in experimental animal models. J Health Sci 53, 245-249. http://dx.doi.org/10.1248/jhs.53.245

Juliani H.R., Koroch A.R., Juliani H.R., Trippi V.S., 1999. Micropropagation of Lippia junelliana (Mold.) Tronc. Plant Cell Tiss Org Cult 59, 175-179. http://dx.doi.org/10.1023/A:1006396531647

Kirtikar K.R., Basu B.D., 1975. Description and medicinal uses of Phyla nodiflora. In: Indian medicinal plants (Singh B., and Singh M.P., ed.) Vol.3, 2nd Ed Jayeed Press, New Delhi, India. pp. 1916-1917.

Leticia C.A., Paiva P.D.O., Paiva R., Garciano H.P., 2010. Growth curve and biochemical analyses of callus of IPE-BRANCO (Tabebuia roseo alba (Ridl.) Sand. Naturalia 33, 45-56.

Lloyd G., McCown B., 1980. Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot tip culture. Intl Plant Prop Soc Proc 30, 421-427.

Murashige T., Skoog F.A., 1962. Revised media for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15, 473-497. http://dx.doi.org/10.1111/j.1399-3054.1962.tb08052.x

Nadkarni A.K., Nadkami K.M., 1954. Indian Materia Medica, Revised and Enlarged. Vol. I, Popular Book Depot. Bombay.

Patel R.M., Shah R.R., 2009. Regeneration of Stevia plant through callus culture. Indian J Pharmaceut Sci 71, 46-50. http://dx.doi.org/10.4103/0250-474X.51954

Ravikanth V., Ramesh P., Diwan P.V., Venkateswarlu Y., 2000. Halleridone and Hallerone from Phyla nodiflora (L.) Greene. Biochem Syst Ecol 28, 905-906. http://dx.doi.org/10.1016/S0305-1978(00)00007-7

Rehman R.U., Israr M., Srivastava P.S., Bansal K.C., Abdin M.Z., 2003. In vitro regeneration of witloof chicory (Cichorium intybus L) from leaf explants and accumulation of esculin. In Vitro Cell Dev Biol - Plant 39, 142-146. http://dx.doi.org/10.1079/IVP2002381

Schenk R.V., Hildebrandt A.C., 1972. Medium and techniques for induction of growth of monocotyledonous and dicotyledonous plant cell cultures. Can J Bot 50, 166-204. http://dx.doi.org/10.1139/b72-026

Shin G., Chil M., Lee D., 2003. Comparative study of the effects of various culture conditions on cell growth and gagaminine synthesis in suspension culture of Cynanchum wilfordii. Biol Pharm Bull 26, 1321-1325. http://dx.doi.org/10.1248/bpb.26.1321

Shukla S., Saluja A.K., Pandya S.S., 2009. In vitro antioxidant activity of aerial parts of Lippia nodiflora Rich. Pharmacol Online 2, 450-459.

Terblanche F.C., Kornelius G., 1996. Essential oil constituents of the Genus Lippia (Verbinaceae) - A literature review. J Ess Oil Res 8, 471-485. http://dx.doi.org/10.1080/10412905.1996.9700673

Yeoman M.M., Yeoman C.L., 1996. Manipulating secondary metabolism in cultured plant cells. New Phytol 134, 553-569 http://dx.doi.org/10.1111/j.1469-8137.1996.tb04921.x

How to Cite
Ahmed, A., Pallela, R., Rao, A., Rao, M., & Mat Taha, R. (1). Optimized conditions for callus induction, plant regeneration and alkaloids accumulation in stem and shoot tip explants of Phyla nodiflora. Spanish Journal of Agricultural Research, 9(4), 1262-1270. https://doi.org/10.5424/sjar/20110904-453-10
Section
Plant physiology