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In vitro elicitation, isolation, and characterization of conessine biomolecule from Holarrhena antidysenterica (L.) Wall. callus and its larvicidal activity against malaria vector, Anopheles stephensi Liston

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Abstract

In vitro elicitation of an important compound conessine has been done in the bark-derived callus culture of Holarrhena antidysenterica (L.) Wall. employing different elicitors. For induction of callus, green bark explants excised from field-grown plants were cultured on MS medium augmented with different concentrations (0, 1, 2.5, 5, and 10 μM) of various growth regulators such as BA, IBA, NAA, and 2,4-D either alone or in combinations. The maximum amount of conessine (458.18 ± 0.89d μg/g dry wt.) was achieved in callus developed on MS medium supplemented with 5 μM BA and 5 μM 2,4-D through HPLC analysis. Elicitation in conessine content in the above callus was achieved employing a variety of organic (phenylalanine, tyrosine, chitosan, tryptophan, casein hydrolysate, proline, sucrose, and yeast extract) as well as inorganic elicitors (Pb(NO3)2, As2O3, CuSO4, NaCl, and CdCl2) in different concentrations. The optimum enhancement in conessine content (3518.58 ± 0.28g μg/g dry wt.) was seen at the highest concentration (200 mg/L) of phenylalanine. The enhancement was elicitor specific and dose dependent. The overall increment of the conessine content was seen in the order of phenylalanine > tryptophan > Pb(NO3)2 > sucrose > NaCl > As2O3 > casein hydrolysate > CdCl2 > chitosan > proline > yeast extract > CuSO4 > tyrosine. The isolation and purification of conessine was done using methanol as a solvent system through column chromatography (CC) and TLC. The isolated compound was characterized by FT-IR, 1H-NMR, and HRMS which confirmed with the structure of conessine. The bioassays conducted with the isolated compound revealed a strong larvicidal activity against Anopheles stephensi Liston with LC50 and LC90 values being 1.93 and 5.67 ppm, respectively, without harming the nontarget organism, Mesocyclops thermocyclopoides Harada, after 48 h of treatment. This is our first report for the isolation and elicitation of conessine in the callus culture of H. antidysenterica.

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Funding

The authors are grateful to the University of Delhi for providing financial assistance in the form of R&D and DST PURSE grants. Dinesh Kumar is indebted to University Grants Commission, New Delhi for the award of JRF.

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Authors and Affiliations

  1. Department of Botany, University of Delhi, Delhi, 110007, India

    Dinesh Kumar & Veena Agrawal

  2. National Institute of Malaria Research, Dwarka, New Delhi, 110077, India

    Gaurav Kumar & Ram Das

  3. Department of Botany, Hindu College, New Delhi, 110007, India

    Ravindra Kumar

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  3. Ram Das
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  4. Ravindra Kumar
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  5. Veena Agrawal
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Correspondence to Veena Agrawal.

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Responsible editor: Philippe Garrigues

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Kumar, D., Kumar, G., Das, R. et al. In vitro elicitation, isolation, and characterization of conessine biomolecule from Holarrhena antidysenterica (L.) Wall. callus and its larvicidal activity against malaria vector, Anopheles stephensi Liston. Environ Sci Pollut Res 25, 6783–6796 (2018). https://doi.org/10.1007/s11356-017-1038-3

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  • DOI: https://doi.org/10.1007/s11356-017-1038-3

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