Invitro regeneration of acorus Calamus – an important medicinal plant

Journal of Current Pharmaceutical Research 2010; 2(1): 36-39Table 1. Effect of different concentration of IAA and IBA on shoot proliferation of Acorus calamus using 1/4 thMS MediaS.N.Concentration(µmol)IAA IBANumber ofexplantsinoculatedNumber ofexplantsproliferated% of shootformationNumber ofshoots/explant1 3.0 2.1 10 2 20 3.52 3.5 2.2 10 3 30 33 4.0 2.4 10 5 50 2.84 4.5 2.4 10 1 10 25 5.0 2.6 10 2 20 26 5.7 2.4 10 9 90 5.6For rooting, well developed shoots were transferred on the MSmedium with IBA alone. The shoot proliferation was done on 1/4 thMS medium containing 5.7µmol IAA and 2.4µmol IBA. Data onpercentage of explants showing shoot proliferation, number ofshoots per culture was recorded and results obtained are shown in aTable 1. Rooted plantlets were removed from the culture tube andagar was washed thoroughly with distilled water. The plantletswere then transferred into small plastic bags containingsand:soil:compost, plantlets were watered every two days for twoweeks, in growth chamber(glass house) where they were exposedto high humidity and temperature for two weeks. The plantletswere finally established in the field for hardening.3. RESULT & DICUSSIONNaturally grown, fresh rhizomes were cut into smallpieces of about 1-3cm, were used as explants for shootregeneration and multiplication.Regeneration of shoot from explant was observed atdifferent hormone concentrations. The highest percentage of shootproliferation was recorded at 1/4thMS+5.7µmol IAA+2.4µmolIBA. Combination of IAA and IBA showed better results. Bestrooting was observed with 1mg/L IBA.The root induced plantletswere successfully transferred to hardening after 30 days andPlantlets produced using rootlets were acclimatized andtransplanted. About 90% of the in vitro Micropropagated plantswere healthy and useful for propagation.4. CONCLUSIONThe sweet flag Acorus calamus rarely produces the seedsand mainly propagated by vegetative means, it has been reported inthe traditional system of medicine for the treatment of dyspepsiaflatulence, cough, fever, piles and asthma. From the present studyit was found that combinations of IAA and IBA responded bettershoot formation and rooting was found best on MS mediumTable 2. Effect of growth regulators on in vitro root regeneration on Acorus calamus shootsS.N. Growth regulators Observation after20 daysobservation after30 daysobservation after40 days1 Half strength MS +0.5mg/L NAA2 Half strength MS +0.5 mg/L IBANo growth No growth No growthNo growth No response No response3 Quarter strength MS +5.7µm IAA + 2.4µmIBANo growthbuff colored rootwas formedManyadventitious rootswere formed4 MS + 1mg/L IBA Root formed Increase in theno.of roots seenWell developedadventious rootsformed38

Journal of Current Pharmaceutical Research 2010; 2(1): 36-39containing 1mg/L IBA alone. So suitable tissue culture techniqueswas established for in vitro shoot proliferation of Acorus calamuswhich can be used for the mass production of diseases free plants,irrespective of season and will serve as an alternative source ofseed materials. In vitro preservation of germplasm is also safemethod to protect species & reducing the risk of natural vagaries.Further estimation of secondary constituents from natural and invitro plants and antimicrobial activity of both natural and invitroplants will be carried out in future.REFERENCESAhmed M.B. et.al, 2007 Standardization of a Suitable Protocol for inVitro Clonal Propagation of Acorus calamus L. an Important Medicinal Plantin Bangladesh. Am-Euras . J. Sci. Res., 2 (2): 136-140.Kokate CK, Purohit AP and Gokhale SB,2003 In:Pharmacognosy,24 thed.Nirali prakashan,pune,pp.379-380.Hettiarachchi A, Fernando KKS and Jayasuriya AHM 1997 Invitropropagation of Wadakha (Acorus calamus L). J.Natn.sci.coun.Sri Lanka25(3):151-157.Raina VK, Srivastava SK, Syamasunder KV. J Flav and frag. 2002;18(1): 18-20.Modhumita, Ghosh. Antifungal properties of haem peroxidase fromAcorus calamus. Ann Bot. 2006; 98(6): 1145-5339