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.
[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
[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
@article{"International Journal of Biological, Life and Agricultural Sciences:57407", author = "Santosh Kumar and Anand Prakash and Kanak Sinha and Anita K Verma", title = "Screening and Evaluation of in vivo and in vitro Generated Insulin Plant (Vernonia divergens) for Antimicrobial and Anticancer Activities", abstract = "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.", keywords = "Anti-cancer, Anti-microbial, EAC mouse cell,
Tissue culture, Vernonia divergens.", volume = "6", number = "10", pages = "922-5", }