Journal of Plant Pathology

, Volume 101, Issue 4, pp 991–995 | Cite as

Microbial contamination in tissue culture of Chlorophytum borivilianum, a rare medicinal herb: identification and prevention

  • Sunil Kumar
  • Amresh Kumar YadavEmail author
  • Chander Prabha
Original Article


When inflorescence axis was used as the explant for micropropagation of Chlorophytum borivilianum, 50% of tubes became contaminated at Stage-I of in vitro culture. Bacteria (Bacillus licheniformis, Micrococcus sp. and Panibacillus sp.) and fungi (Alternaria sp., Aspergillus sp., Cladosporium sphaerospermum and Fusarium sp.) were the most common contaminants.The effectiveness of synthetic antimicrobial compounds in inhibiting the growth of contaminants was compared. Five antibiotics, penicillin (PE), norfloxacin (NF), tobramycin (TB), gatifloxacin (GT) and ofloxacin (OF), at 1%–15%, and four fungicides, Bavastin (BV), Captan (CA), Fluconazole (FL) and Trichoderma biofungicide (TV), at 0.5–2%, were tested on the bacterial and fungal isolates respectively. GT (5%) and OF (8%) inhibited 100% growth of bacteria, but the former was more efficient. BV and CA at 1–2% appeared to be the most effective fungicides. MIC of antibiotics was higher (8–10%) than MIC of fungicides (1–2%). The two most effective antibiotics (GT, OF) and fungicides (BV, CA) were also evaluated in vitro for phytotoxicity. The observed MPCs for GT, OF, BV and CA were 4%, 9%, 1% and 1.5% respectively. They were then tested in combination at their respective MICs in amended MS media and culture survival was recorded at Stage-I. Ninety per cent of cultures survived with GT (4%) + BV(1%) combination, while the combination of GT and CA was highly toxic. Enhancement in culture survival by 40% resulted from synergistic or additive effects of GT and BV.


Micropropagation Antimicrobial compounds MIC MPC Phytotoxicity 



We are grateful to the Head, Department of Botany, Patna University, Patna, Bihar, India for providing laboratory facilities (Plant Tissue Culture and Micro Biology) and others technical supports.

Author’s contribution

All authors contributed equally to research work.

Compliance with ethical standards

Conflict of interest

We have no conflict of interest for publication of this content.


  1. Altan F, Burun B, Sahin N (2010) Fungal contaminants observed during micropropagation of Lilium candidum L. and the effect of chemotherapeutic substances applied after sterilization. Afr J Biotechnol 9:991–995CrossRefGoogle Scholar
  2. Andrew JM (2001) Determination of minimum inhibitory concentration. J Antimicrob Chemother 48:5–16CrossRefGoogle Scholar
  3. Barnet HL, Hunter BB (1998) Genera of Imperfect. Illustrated Fungi, 4th edn. American Phytological Society, St. PaulGoogle Scholar
  4. Borum DF, Sinclair JB (1965) Evidence for systemic fungicide protection against Rhizoctonia solani with Vitavax in cotton seedlings. Phytopathology 58:979–980Google Scholar
  5. Chai KL, Dayang AWA, Lau CY, Sim SL (2010) Control of in vitro contamination of explants from field-grown dabai (Canarium odontophyllum Miq.) trees. Asia Pac J Mol Biol Biotechnol 18:115–118Google Scholar
  6. Claus GW (1995) Understanding microbes, 4th edn. WH Freeman and Company, New York, p 547Google Scholar
  7. Desai S, Desai P, Mankad M, Patel A, Patil G, Narayanan S (2018) In Vitro response of nine different genotype of “Safed Musli” (Chlorophytum borivilianum) using grown shoot bud as an explant. Int J Pure Appl Biosci 6:1414–1420CrossRefGoogle Scholar
  8. Garcia PC, Rivero RM, Ruiz JM, Romero L (2003) The role of fungicides in the physiology of higher plants: implications for defense responses. Bot Rev 69:162–172CrossRefGoogle Scholar
  9. Gilman CJ (1957) A manual of soil fungi. Iowa State College Press, Iowa, p 450Google Scholar
  10. Gradelski E, Kolek B, Bonner D, Tomc JF (2002) Bactericidal mechanism of gatifloxacin compared with other quinolones. J Antimicrob Chemother 49:185–188CrossRefGoogle Scholar
  11. Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST (1994) Bergey’s manual of determinative bacteriology, 9th edn. Lippincot Williams & Wilkins, PhiladelphiaGoogle Scholar
  12. Jena RC, Samal KC (2011) Endogenous microbial contamination during in vitro culture of sweet potato [ Ipomea batatas (L.) Lam]: identification and prevention. J Agric Technol 7:1725–1733Google Scholar
  13. Khan AM, Qureshi RA, Ullah F, Gilani SA (2011) Phytotoxic effects of selected medicinal plants collected from Margalla Hills, IslamabadPakistan. J Med Plants Res 5:4671–4675Google Scholar
  14. Kopila A, Anuradha KN, Prabhu Suchitra N (2018) Pharmacognostic evaluation and HPTLC finger printing of rhizome of Chlorophytum borivilianum Sant. and F. from Nepal. Pharm J 10:963–968Google Scholar
  15. Kotra LP, Haddad J, Mobashery (2000) Aminoglycosides: prospective on mechanism of action and resistance and strategies to counter resistance. Antimicrob Agents Chemother 44:3249–3256CrossRefGoogle Scholar
  16. Leifert C, Cassells AC (2001) Microbial hazards in plant tissue culture and cell cultures. In-vitro cell Dev Bio Plant 37:133–138CrossRefGoogle Scholar
  17. Mathew A, Malathy RM (2008) The evidence of mycorrhizal fungi and dark septate endophytes in roots of Chlorophytum borivillianum. Acta Botanica Croatica 67:91–96Google Scholar
  18. Msogoya T, Kanyagha H, Mutigitu J, Kulebelwa M, Mamiro D (2012) Identification and management of microbial contaminants of banana in-vitro cultures. J Appl Biosci 55:3987–3994Google Scholar
  19. Nagy JK, Sule S, Sampaio JP (2005) Apple tissue culture contamination by Rhodotorula spp.: identification and prevention. In Vitro Cell Dev Biol-Plant 41:520–524CrossRefGoogle Scholar
  20. Narasimham KRSL, Ravuru BK (2003) A note on some endangered medicinal plants as NTFPs of eastern ghats, Andhra Pradesh. EPTRI-ENVIS Newslett 9:11–12Google Scholar
  21. Nelson PE, Toussoun TA, Marasas WFO (1982) Fusarium spp. An Illustrated Manual of Identification. Pennsylvania UniversityPress, Philadelphia, p 216Google Scholar
  22. Panathula CS, Mahadev MDN, Naidu CV (2014) The stimulatory effect of the antimicrobial agents Bavistin, cefotaxime and kanamycin on in-vitro plant regeneration of Centella asiatica (L.) - an important anti-jaundice medicinal plant. Am J Plant Sci 5:279–285CrossRefGoogle Scholar
  23. Panchal H, Ingle S (2011) Isolation and characterization of endophytes from the root of medicinal plant Chlorophytum borivilianum (Safed musli). J Adv Devl Res 2:205–209Google Scholar
  24. Shailja VV, Himabindu V, Anuradha K, Ananad T, Lakshmi V (2004) In vitro activity of gatifloxacin against gram negative clinical isolates in a TertiaryCareHospital. Indian J Med Microbiol 22:222–225Google Scholar
  25. Sharma PC, Sanjeja A, Jain S (2008) Norfloxacin: a therapeutic review. Int J Chem Sci 6:1702–1713Google Scholar
  26. Sharma R, Thakur G, Sanodiya BS, Pandey M, Bisen PS (2012) Saponin: a wonder drug from Chlorophytum species. Glob J Res Med Plants Indigenous Med 1:503–515Google Scholar
  27. Sundaram S, Dwivedi P, Purwar S (2011) Antibacterial activities of crude extracts of Chlorophytum borivilianumto bacterial pathogens. J Med Plants Res 5:343–347CrossRefGoogle Scholar
  28. Tripathi RK, Ram S (1982) Induction of growth and differentiation of carrot callus cultures by Carbendazim and Benzimidazole. Indian J Exp Biol 20:674–677Google Scholar
  29. Vincent JM (1947) Distortion of fungal hyphae in presence of certain inhibitors. Nature 45:398–404Google Scholar
  30. World Health Organization (2013) Strategies for global surveillance of antimicrobial resistance: Report of a technical consultation (WHO/HSE/PED/2013.10358), Geneva. 2012/en/index.html. Accessed 6 Jan 2014
  31. Yadav AK, Ambasta SK, Prasad SK, Trvedi MP (2018) In-vitro evaluation of antibacterial property of Catharanthus roseus (Linn.) G. Don. Var. “rosea” and “alba”. Int J Pharm Pharm Sci 10:55–58CrossRefGoogle Scholar

Copyright information

© Società Italiana di Patologia Vegetale (S.I.Pa.V.) 2019

Authors and Affiliations

  1. 1.Department of BotanyPatna UniversityPatnaIndia

Personalised recommendations