Screening and quantification of an antiseptic alkylamide, spilanthol from in vitro cell and tissue cultures of Spilanthes acmella Murr.

Department of Biotechnology, Indian Institute of Technology - Guwahati, Guwahati 781039, Assam, India
Industrial Crops and Products (Impact Factor: 3.21). 03/2012; 36(1):321–328. DOI: 10.1016/j.indcrop.2011.10.029

ABSTRACT The study revealed, for the first time, accumulation of spilanthol, an antiseptic alkylamide, in in vitro cultures of Spilanthes acmella Murr., a medicinal plant of immense commercial value. To achieve this, in vitro shoots were regenerated via direct organogenesis from leaf-disc explants of Spilanthes. Shoots were induced in the presence of N6-benzylaminopurine (BAP) alone or in combination with either α-naphthalene acetic acid (NAA) or Indole-3-acetic acid (IAA) in Murashige and Skoog medium. The best treatment for shoot regeneration was MS + BAP (5.0 μM) + IAA (5.0 μM), which promoted adventitious shoot proliferation in >82% cultures with an average of 5.3 shoots per explant. Regenerated shoots rooted spontaneously with a frequency of 100% on half strength MS medium (major salts reduced to half strength) containing 50 g l−1 sucrose. The plantlets were acclimatized successfully with 90% survival rate. Additionally, ploidy stability of the regenerated plants was assessed by flow cytometry which showed that all investigated plants had the similar ploidy as that of the mother plant. For spilanthol identification, peaks eluted from HPLC were analyzed by mass spectrometry with its characteristic fragmentation pattern. For quantification studies, calibration curve was generated, which revealed a higher amount of spilanthol content (3294.36 ± 12.4 μg/g DW) in the leaves of in vitro plants compare to those of in vivo plants (2703.66 ± 9.6 μg/g DW of spilanthol). An efficient multiplication frequency, ploidy stability and enhanced spilanthol accumulation ensure the efficacy of the protocol developed for this industrially important medicinal plant.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Acmella oleraceae (L.) R.K. Jansen (Asteraceae) is a therapeutically important medicinal plant used in traditional systems of medicine. Present study was undertaken to compare the total phenolic contents (TPC), total flavonoid contents (TFC), total antioxidant capacity (TAC) and Thin Layer Chromatography (TLC) profiles of areal parts and callus extracts of A. oleraceae grown under different growing systems (field grown, hydroponically grown and callus culture. Callus established in Murasige and Skoog (MS) medium, areal parts of field grown and hydroponically grown plants were extracted in 80% methanol. TPC, TFC and TAC were carried out by using colorimetric Folin-Ciocalteu method, aluminum nitrate method and Ferric Reducing Antioxidant Power (FRAP) assay respectively. TLC profiles were developed using established protocol. The best callus growth and the highest mean callus weight were observed in leaf explants established in MS medium supplemented with 2 mg/L BA and 1 mg/L IBA. Comparatively higher TPC (11.45 ± 0.17), TFC (12.33 ± 0.92) and TAC (10.27 ± 0.28) were observed in hydroponically grown plants. Order of TPC, TFC and TAC were increased as callus<field grown plants< hydroponically grown plants. The presence of higher TPC, TFC and TAC in hydroponically grown plants opens a new window for use of hydroponic system for growing of A. oleraceae for better secondary metabolites content.
    07/2014; 2(2):163-167.
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Plants have been extensively used for medicinal purposes and human disease management since early ages, due to presence of valuable chemical identities. Present study was carried out to identify chemical identities and the cytotoxicity of Spilanthes paniculata Wall. ex DC. which belongs to family Asteraceae. This plant is commonly known as 'Tooth-ache plant' due to the presence of anesthetic properties. A mother stock has been maintained for experiments. Leaves obtained from green-house grown plants and in vitro callus cultures of 3-weeks-old were used in this study. In vitro callus induction was optimized with leaf disc explants grown on MS medium supplemented with 0.15 mg/L of NAA and 1.50 mg/L of BA. GC-MS analysis of ethyl acetate extracts of leaf and callus samples confirmed the existence of the key chemical compound spilanthol in both samples, with significantly higher amount in fresh leaves. Cytotoxicity tests confirmed the toxicity of hexane extracts of leaf and callus samples against late third/ early fourth instar larvae of Aedes aegypti and nuaplii of Artemia salina (100% mortality with 800 µg/mL). Leaf extracts were found to be highly toxic (70% mortality with 100 ppm for A. aegypti and 100% mortality with 800 µg/mL for nuaplii of A. salina) to both test organisms than callus extract.
    International Journal of Herbal Medicine. 08/2013; 1(3):135-141.
  • Source
    British Biotechnology Journal. 01/2013; 3(3):405-415.


Available from
May 16, 2014