Isolation, quantification and anti‑microbial screening of flavonoids from callus of Vitex negundo Linn

Authors

  • Keerti Gautam
  • Padma Kumar

DOI:

https://doi.org/10.22377/ijgp.v7i4.340

Abstract

Context: Natural habitats for medicinal plant are disappearing fast and together with environmental instabilities, it is increasingly difficult to acquire plant‑derived compounds. This has prompted scientists to consider the possibilities of using cell cultures as an alternative supply for the production of plant natural products.

Aim: The present investigation deals with the establishment of callus cultures from leaf explants of Vitex negundo, to extract flavonoids from the raised callus and to assess their anti‑microbial potential. Materials and Methods: Callus culture of Vitex negundo has been established on Murashige and Skoog’s medium supplemented with 2, 4‑D using leaf explants. Growth index (GI) was calculated for the tissues obtained after 2, 4, 6 and 8 weeks, respectively. Calli of 2, 4, 6 and 8 weeks old were used for extraction of total flavonoids (free and bound) using standard method. Flavonoid extract of 6 weeks old callus with maximum GI and maximum flavonoid content was then tested against Staphylococcus aureus, Agrobacterium
tumefaciens (bacteria), Candida albicans (yeast) for their anti‑microbial potential. Minimum inhibitory concentration, minimum bactericidal/fungicidal concentrations and total activity were also evaluated. Results: GI was found maximum in 6 weeks old tissue (0.76). Total flavonoids from 6 week old tissue showed maximum zone of inhibition against A. tumefaciens. Kaempferol was identified from total flavonoids of 6 week old callus through, thin layer chromatography (TLC), preparative TLC, Melting point and Infra red spectral studies. Content of kaempferol in 6‑week‑old calli was also estimated using standard Spectrophotometric method, which came out to be 54.25 μg/mg d.wt (dry weight) of calli. Conclusion: The protocol of callus induction of V. negundo opens new vistas that could facilitate phytochemical production, extraction of pharmaceuticals and their screening against different human pathogens
from the callus without harvesting the plant itself.
Key words: Anti‑microbial potential, callus, flavonoids, Vitex negundo

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References

Baris O, Gulluce M, Sahin F, Ozer H, Killic H, Ozkan H, et al.

Biological activities of the essential oil and methanol extract of

Achillea Biebersteinii Afan (Asteraceae). Turk J Biol 2006;30:65‑73.

Mulabagal V, Tsay HS. Plant cell cultures an alternative and

efficient source for the production of biologically important

secondary metabolites. Int J Appl Sci Eng 2004;2:29‑48.

Di Cosmo F, Misawa M. Plant cell and tissue cultures: Alternatives

for metabolite production. Biotechnol Adv 1995;13:25‑53.

Rao RS, Ravishankar GA. Plant cell cultures: Chemical factories

for secondary metabolites. Biotechnol Adv 2002;20:101‑53.

Zheng CJ, Tang WZ, Huang BK, Han T, Zhang QY, Zhan H,

et al. Bioactivity‑guided fractionation for analgesic properties

and constituents of Vitex negundo L. seeds. Phytomedicine

;16:560‑7.

Zheng CJ, Huang T, Han QY, Zhang H, Rahman K, Qin LP.

Antinocieceptive activities of the liposoluble fraction from Vitex

negundo seeds. Pharm Biol 2010;48:651‑8.

Yunos NM, Ali RM, Kean OB, Abas R. Cytotoxicity Evaluations

on Vitex negundo Antiinflammatory extracts. Malays J Sci

;24:213‑7.

Jana GC, Baliga MS. The evaluation of nitric oxide scavenging

activity of certain Indian medicinal plants in vitro: A preliminary

study. J Med Food 2004;7:343‑8.

Dharmasiri MG, Jayakody JR, Galhena G, Liyanage SS,

Ratnasooriya WD. Anti‑inflammatory and analgesic activities

of mature fresh leaves of Vitex negundo. J Ethnopharmacol

;87:199‑206.

Tondon VR, Gupta RK. Anti‑inflammatory activity and mechanism

of action of Vitex negundo Linn. Int J Pharmacol 2006a;2:303‑8.

Das S, Praveen S, Kundra CP, Pereira BM. Reproduction in male

rats is vulnerable to treatment with the flavonoid rich seed extracts

of Vitex negundo. Phytother Res 2004;18:8‑13.

Chandramu C, Rao DM, Krupanandam DG, Reddy DV. Isolation,

characterization and bilolgical activity of betulinic acid and ursolic

acid from Vitex negundo L. Phytother Res 2003;17:129‑34.

Sahayaraj K. Antifeedant effect of some plant extracts on the Asian

armyworm, Spodoptera litura (Fabricius). Curr Sci 1998;74:523.

Tiwari OP, Tripathi YB, Antioxidant properties of different fractions

of Vitex negundo Linn. Food Chem 2007;100:1170‑6.

Vishal T, Gupta RK. Effect of Vitex negundo on oxidative stress.

Indian J Pharmacol 2005;37:38‑40.

Rooban B, Lija Y, Biju P, Sasikala V, Sahasranamam V, Abraham A.

Vitex negundo attenuates calpain activation and cataractogenesis

in selenite models. Exp Eye Res 2009;88:575‑82.

Villasenor IM, Lamadrid MR. Comparative anti hyperglycemic

potentials of medicinal plants. J Ethnopharmacol 2006;104:129‑31.

Diaz F, Chavez D, Lee D, Mi Q, Chai HB, Tan GT, et al. Cytotoxic

flavones analogues of vitexicarpin, a constituent of the leaves of

Vitex negundo. J Nat prod 2003;66:865‑7.

Yang LK, Yen K, Kiso Y, Hikino H. Antihepatotoxic actions of

Formosan plant drugs. J Ethanopharmacol 1987;19:103‑10.

Tondon VR, Khajuria V, Kapoor B, Kour D, Gupta S. Hepatoprotective

activity of Vitex negundo leaf extract against anti‑tubercular drugs

induced heptotoxicity. Fitoterapia 2008;79:533‑8.

Tasduq SA, Kaiser PJ, Gupta BD, Gupta VK, Johri RK. Negundoside,

an irridoid glycoside from leaves of Vitex negundo, protects human

liver cells against calcium‑mediated toxicity induced by carbon

tetrachloride. World J Gastroenterol 2008;14:3693‑709.

Raj PV, Chandrasekhar HR, Vijayan P, Dhanarj SA, Rao CM,

Rao JV, et al. In vitro and in vivo hepatoprotective effect of Vitex

negundo leaves. Pharmacol Online 2008;3:281‑95.

Adnaik RS, Pai PT, Mule SN, Nikawade NS, Magdum CS. Laxative

activity of Vitex negundo Linn. Leave. Asian J Exp Sci 2008;22:159‑60.

Ravishankar B, Shukla V. Indian systems of Medicine: A brief

profile. Afr J Tradit Complement Altern Med 2007;4:319‑37.

Karunamoorthi KS, Ramanujan S, Rathinasamy R. Evaluation of

leaf extract of Vitex negundo L. (family: Verbenaceae) against larvae

of Culex tritaeniorhynchus and repellent activity on adult vector

mosquitoes. Parasitol Res 2008;103:545‑50.

Hebbalkar DS, Hebbalkar GD, Sharma RN, Joshi VS, Bhat VS.

Mosquito repellant activity of oils from Vitex negundo Linn. Leaves.

Indian J Med Res 1992;95:200‑3.

Subramanian SS, Nagarjan S. Flavonoids of the seeds of Crotolaria

retusa and Crotolaria striata. Curr Sci 1969;38:65.

Naghski J, Fenske CS Jr, Couch JF. Use of paper chromatography

for the estimation of quercetin in rutin. J Am Pharm Assoc

;40;613‑6.

Kariyone T, Hashimoto Y, Kimura M. Microchemical studies on

plant components. IX. Distribution of flavonoids in plants by paper

chromatography. J Pharm Soc (Japan) 1953;73:253‑6.

Mabry TJ, Marjhan KR, Thomas MB, editors. In: The Systematic

Identification of Flavonoids. Berlin: Springer‑Verlag; 1970; p. 119.

Andrews JM. BSAC standardized disc susceptibility testing

method. J Antimicrob Chemother 2001;4:43‑57.

Gould JC, Bowie JH. The determination of bacterial sensitivity of

antibiotics. Edinb Med J 1952;59:178‑99.

Jain N, Sharma M. Broad spectrum antimycotic drug for the

treatment of ringworm infections in human beings. Curr Sci

;85:30‑4.

Basri DF, Fan SH. The potential of aqueous and acetone extracts

of galls of Quercus infectoria as antibacterial agents. Indian J

Pharmacol 2005;37:26‑9.

Eloff JN. Quantifying the bioactivity of the plant extracts during

screening and bioassay‑guided fractionation. Phytomedicine

;11:370‑1.

Usha PK, Sailas B, Mohanan KV, Raghu AV. An Efficient micro

propagation system for Vitex negundo L., an important woody

aromatic medicinal plant, through shoot tip culture. Res J Bot

;2:102‑7.

Jawahar M, Ravipaul S, Jeyaseelan M. In vitro regeneration of Vitex

negundo L.: A multipurpose woody aromatic medicinal shrub.

Plant Tissue Cult Biotech 2008;37‑42.

Chowdhury FB, Azam SF, Hassan M, Jahan FI, Chowdhury AR,

Seraj S, et al. Studies with callus induction of Vitex negundo:

An aromatic medicinal plant. Am Eurasian J Sustain Agric

;5:6‑14.

Gautam K, Kumar P. Extraction and pharmacological evaluation

of some extracts of Vitex negundo Linn. Int J Pharm Pharm Sci

;4:132‑7.

Rose MC, Cathrine L. Preliminary phytochemical screening

and antibacterial activity on Vitex negundo. Int J Curr Pharm Res

;3:99‑101.

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