Screening and Evaluation of in vivo and in vitro Generated Insulin Plant (Vernonia divergens) for Antimicrobial and Anticancer Activities

Vernonia divergens Benth., commonly known as “Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the leaves of the plant, boiled in water are successfully administered to a large number of diabetic patients. The present study evaluates the putative anti-diabetic ingredients, isolated from the in vivo and in vitro grown plantlets of V. divergens for their antimicrobial and anticancer activities. Sterilized explants of nodal segments were cultured on MS (Musashige and Skoog, 1962) medium in presence of different combinations of hormones. Multiple shoots along with bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA. Micro-plantlets were separated and sub-cultured on the double strength (2X) of the above combination of hormones leading to increased length of roots and shoots. These plantlets were successfully transferred to soil and survived well in nature. The ethanol extract of plantlets from both in vivo & in vitro sources were prepared in soxhlet extractor and then concentrated to dryness under reduced pressure in rotary evaporator. Thus obtainedconcentrated extracts showed significant inhibitory activity against gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa but no inhibition was found against gram positive bacteria. Further, these ethanol extracts were screened for in vitro percentage cytotoxicity at different time periods (24 h, 48 h and 72 h) of different dilutions. The in vivo plant extract inhibited the growth of EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50, 25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in vitro origin, the inhibition was found against EAC cell lines even at 48h. During spectrophotometric scanning, the extracts exhibited different maxima (ʎ) - four peaks in in vitro extracts as against single in in vivo preparation suggesting the possible change in the nature of ingredients during micropropagation through tissue culture techniques.

Authors:

• Santosh Kumar
• Anand Prakash
• Kanak Sinha
• Anita K Verma

Keywords:

• Anti-cancer
• Anti-microbial
• EAC mouse cell
• Tissue culture
• Vernonia divergens.

References:

[1] M. M. Cowman, "Plant products as antimicrobial agents," Clinical
Microbial Review, vol, 12, pp. 564-582, 1999.
[2] S. S. Gyang, D. D. Nyam and E. N. Sokomba, "Hypoglycaemic activity
of Vernonia amygdalina (chloroform extract) in normoglycaemic and
alloxan-induced hyperglycaemic rats," Journal of Pharmacy and
Bioresources, vol. 1, pp. 61-66, 2004.
[3] G. V. Gill, S. Redmond, F. Garrott, R. Paisey, "Diabetes and alternative
medicine: a case for concern," Diabetic Medicine, vol. 22, pp. 210-213,
1994.
[4] G. P. Lease and G. Williams, "Complementary and conventional
medicine in clinical diabetes," International diabetes digest, vol. 6, pp.
675-676, 1994.
[5] P. G. G. Odeigah, I. A. Taiwo, E. O. Akomolafe and D. O. Durojaiye,
"Hypoglycaemic actions of medicinal plants with tolbutamide in the
albino rat," Diabetes international, vol. 9, pp. 71-73. 1999.
[6] I. A. Taiwo, P. G. C. Odeigah, L. A. Ogunkanmi, "The glycemic effects
of Vernonia amygdalina and V. tenoreana with tolbutamide in rats and
the implications for the treatment of diabetes mellitus," Journal of
Scientific Research and Development, vol. 11, pp. 122-130, 2008.
[7] T. Murashige and F. A. Skoog, "Revised medium for rapid growth and
bioassays with tobacco tissue culture," Physiol Plantarum, vol. 15, pp.
473-497, 1962.
[8] T. Mosmann, "Rapid colorimetric as say for cellular growth and
survival, application to proliferation and cytotoxicity assays," Journal of
Immunoogical Methods, vol. 65, pp. 55-63, 1983
[9] C. Parez, M. Paul and P. Bazerque, "Antibiotic assay by agar well
diffusion method," Acta. Biol. Med. Exp, vol. 15, 113-115, 1990.
[10] P. S. Reddy, P. Rodrigues and R. Rajasekharan, "Shoot organogenesis
and mass propagation of Coleus forskohlii from leaf derived callus,"
Plant Cell Tissue Organ Culture, vol. 66: 183-188, 2001
[11] S. Shrivastana and M. Banerjee, "In vitro clonal propagation of physic
nut (Jatropha curcas L.), Influence of additives," Int Integrative Biol.
Vol. 3,73-79, 2008
[12] S. Kumar, A. Prakash, R. Vohra and A. K.. Verma, "Evaluation of
anticancer activities of ethanolic extracts of in vivo and in vitro grown
Coleus forskohlii Berq." Perspective in Cytology and Genetics, vol. 15,
pp. 19-30, 2011.
[13] V. Sharma, S. A. Mallick and A. K. Tiku, "Anticancer activity of devil
tree (Alstonia scholaris Linn.) leaves on human cancer cell lines,"
Indian Journal of Agricultural Biochemistry, vol. 23, pp. 63-65, 2010.
[14] Kandukuri and M. A. Singara, "Antimicrobial activity of five flower
extracts against three pathogenic bacteria,- Nature Environment and
Pollution Technology, vol. 9, pp. 73-76, 2010