RAPID IN VITRO CALLOGENESIS AND PHYTOCHEMICAL SCREENING OF LEAF, STEM AND LEAF CALLUS OF MUSSAENDA FRONDOSA LINN. - A MEDICINAL PLANT.


Manasa Dj, Chandrashekar Kr, Bhagya N

Abstract


Objective: To standardise the protocol for rapid callogenesis in Mussaenda frondosa L. using leaf explants. Qualitative and quantitative phytochemical analysis of leaf, stem and callus cultures.

Methods: The leaf explants were inoculated onto MS medium supplemented with varying concentrations of growth regulators such as 2, 4 - D, NAA, BAP, Kn for the induction of callus. Qualitative and quantitative analysis of total phenol, flavonoids and alkaloids contents of leaf, stem and callus were tested by standard methods.  The antioxidant activities were investigated using DPPH radical scavenging method and reducing power assay. The anti - inflammatory activity was evaluated by membrane stabilizing activity.

Results: Pale green, healthy, friable and fast growing callus was obtained on the medium enriched with NAA (2mg/l) + Kn (4mg/l). Quantitative determination showed the highest concentration of total phenolics in the methanolic extract of in vitro grown callus (10 ± 1.1 mg of GA/g of extract), flavonoids in methanolic stem extract (137±1.6 mg of Quercitin/g of extract) and alkaloids in methanolic extract of leaf (118.3±1.5 mg/10g of extract). The methanolic leaf extract exhibited highest free radical scavenging activity with IC50 value of 40.6±10.06 μg/ml. The highest membrane stabilizing activity was shown by chloroform extract of the leaf (66.02%).

Conclusion: The present preliminary phytochemical and pharmacological analysis may form the basis for drug development in future using callus cultures of M. frondosa.

 

 

 


Keywords


Mussaenda frondosa; antioxidant; anti – inflammatory; in vitro culture; phenolics, flavonoids, alkaloids.

| PDF |

References


Cragg GM, Newman DJ. Natural product drug discovery in the next millennium. Pharm Biol 2001;39 Suppl 1:8-17.

Sirigiri CK, Kokkanti M. In vitro analysis of cytotoxicity and 5- lipoxygenase inhibition activity by using callus extract of Biophytum sensitivum (Linn) DC. Int J Pharm Sci Rev Res 2014;34:215-8.

Havkin FD, Leustek T. Plant tissue culture for production of secondary metabolites. Food Technol 1997;51(11):56-61.

Sirigiri CK, Kokkanti M, Challa SR. In vitro pharmacological investigations of using leaf callus extracts of Canthium parviflorum Lam. Br Biotechnol J 2014;4(3):325-38.

Alfermann AW, Petersen M. Natural products formation by plant cell biotechnology: Results and perspectives. Plant Cell Tissue Organ Culture 1995;43:199-205.

Verpoorte R, Contin A, Memelink J. Biotechnology for the production of plant secondary metabolites. Phytochem Rev 2002;1(1):13-25.

Maneechai S, De-Eknamkul W, Umehara K, Noguchi H, Likhitwitayawuid K. Flavonoid and stilbenoid production in callus cultures of Artocarpus lakoocha. Phytochemistry 2012;81:42-9.

Loredo-Carrillo SE, Santos-Diaz M, Layva E. Establishment of callus from Pyrostegia venusta (Ker Gawl) Miers and effect of abiotic stress on flavonoids and sterols accumulation. J Plant Biochem Biotechnol 2013;22(3):312-8.

Sahin G, Verma SK, Gurel E. Calcium and magnesium elimination enhances accumulation of cardenolides in callus cultures of endemic Digitalis species of Turkey. Plant Physiol Biochem 2013;73:139-43.

Szopa A, Ekiert H. Production of biologically active phenolic acids in Aronia melanocarpa (Michx.) Elliott in vitro cultures cultivated on different variants of the Murashige and Skoog medium. Plant Growth Regul 2014;72(1):51-8.

Duangporn P, Siripong P. Effect of auxin and cytokinin on phyllanthusol a production by callus cultures of Phyllanthus acidus Skeels. Am Eurasian J Agric Environ Sci 2009;5(2):258-63.

Huxley AM, Levy M. The New Royal Horticultural Society Dictionary of Gardening. Vol. 3. New York: Groves Dictionaries, Inc.; 1999. p. 271-2.

Kirtikar KR, Basu BD. Indian Medicinal Plants. 2nd ed. Dehradun: International Book Distributors; 1987.

Jain SK. Dictionary of Indian Folk Medicine and Ethno Botany. New Delhi: Deep Publication; 1991.

Roshishaliya AP, Kumar U, Upwar N, Patel N. Diuretic study of ethanolic and aqueous extract of Mussaenda frondosa Linn. in normal rats. Available from: http://www.scientificipca.org.

Sambrekar SN, Patil PA, Kangralkar VA, Protective activity of Mussaenda frondosa Linn leaf extract against paracetamol induced hepatic damage in wistar rats. J Pharm Res 2010;3(4):711.

Wesley JJ, Jeyaananath J, Mohammed AA, Ravikumar K. Hypolipidemic effect of methanolic extract of Mussaenda frondosa Linn. leaves in high fat diet fed rats. J Pharm Res 2009;2(4):579-81.

Koul S, Chaudhary A. Radical scavenging and antistress activity of Mussaenda frondosa Linn roots (Rubiaceae). Pharmacologyonline 2011;1:1091-7.

Siju EN, Rajalakshmi GR, Hariraj N, Sreejith KR, Sudhakaran S, Muneer EK, et al. In-vitro anthelmintic activity of Mussaenda frondosa. Res J Pharm Technol 2010;3(1):151-3.

Anonymous. Dictionary of Chinese Traditional Medicine. Shanghai: Jiangsu New Medical College, Shanghai Science and Technology Press; 1986. p. 176.

Ranarivelo Y, Skaltsounis AL, Andriantsiferana M, Tillequin F. Glycosides from Mussaenda arcuata Lam. ex Poiret leaves. Ann Pharm Fr 1990;48(5):273-7.

Biswanath D, Sudhan D, Santanu M, Shiho A, Nariko S, Yoshihiro H. Chemical constituents of Mussaenda incana. Indian J Chem 2005;44B:2362-6.

Michael D. Medicinal Plants in the South Pacific. Western Pacific Series No. 19. Geneva: WHO Regional Publications; 1998. p. 125.

Das P. Mass cloning of rose and Mussaenda, popular garden plants, via somatic embryogenesis. Hortic Sci Prague Czech Republ 2010;37:70-8.

Bhat KG. Flora of Udupi. Karnataka, India: Indian Naturalist (Regd.); 2003.

Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 1962;15:467-97.

Harborne JB. Phytochemical methods. A Guide to Modern Techniques of Plant Analysis. 2nd ed. London: Chapman and Hall; 1973.

Sofowora A. Medicinal Plants and Traditional Medicine in Africa. Ibadan, Nigeria: Spectrum Books Ltd.; 1993. p. 191-289.

Ikram EH, Eng KH, Jalil AM, Ismail A, Idris S, Azlan A, et al. Antioxidant capacity and total phenolic content of Malaysian underutilized fruits. J Food Compos Anal 2009;22(5):388-93.

Sreevidya N, Mehrotra S. Spectrophotometric method for estimation of alkaloids precipitable with Dragendorff’s reagent in plant materials. J AOAC Int 2003;86(6):1124-7.

Zishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging affects on superoxide radicals. Food Chem 1999;64:555-9.

Oyaizu M. Studies on products of browning reactions: Antioxidative activities of products of browning reaction prepared from glucosamine. Jpn J Nutr 1986;44:307-15.

Shinde UA, Phadke AS, Nair AM, Mungantiwar AA, Dikshit VJ, Saraf MN. Studies on the anti-inflammatory and analgesic activity of Cedrus deodara (Roxb.) Loud. wood oil. J Ethnopharmacol 1999;65(1):21-7.

Lubaina AS, Murugan K. Effect of growth regulators in callus induction, plumbagin content and indirect organogenesis of Plumbago zeylanica. Int J Pharm Pharm Sci 2012;4(1):334-6.

Bhagya N, Chandrashekar KR. Evaluation of plant and callus extracts of Justicia gendarussa burm. F. for phytochemicals and antioxidant activity. Int J Pharm Pharm Sci 2013;5(2):82-5.

Namde H, Minal W. Callus induction studies and active components and antioxidant activity investigation from leaves and callus of Tecoma stans L. Juss. ex Kunth. Res J Pharm Biol Chem Sci 2014;5(2):604-10.

Thalwal S, Pannu J, Gupta A. Comparison of antimicrobial activity and phytochemical constituents of in vivo and in vitro grown Amaranthus spinosus plants. Int J Pharm Pharm Sci 2013;5(3):703-7.

Kubmarawa D, Ajoku GA, Enworem NM, Okorie DA. Preliminary phytochemical and antimicrobial screening of 50 medicinal plants from Nigeria. Afr J Biotechnol 2007;6(14):1690-6.

Mensah JK, Okoli RI, Ohaju-Obodo JO, Eifediyi K. Phytochemical, nutritional and medical properties of some leafy vegetables consumed by Edo people of Nigeria. Afr J Biotechnol 2008;7(14):2304-9.

Tanwer BS, Vijayvergia R. Phytochemical evaluation and quantification of primary metabolites of Alangium salviifolium. Int J Pharm Bio Sci 2010;1(3):1-6.

Bhagya N, Chandrashekar KR, Muralidharan K, Amarnath CH. Phytochemical analysis and antioxidant activity of in vitro cultured stem callus of Cyclea peltata. J Trop Med Plants 2012;13(2):117-23.

Jeong GT, Woo JC, Park DH. Effect of plant growth regulators on growth and biosynthesis of phenolic compounds in genetically transformed hairy roots of Panax ginseng C. A. Meyer. Biotechnol Bioprocess Eng 2007;12:86-91.

Taj Al-Deen AM. Application tissue culture technique on some medicinal and aromatic plants used in Yemen. Ph-D Thesis Faculty of Science University of Sana’a; 2010.

Wani M, Sharma A, Deshpande J, Mathew S, Khetmalas MBQualitative phytochemical screening of three indigenous medicinal plants. Res J Pharm Biol Chem Sci 2013;4(1):287.

Umesh TG. In vitro callus induction and antioxidant potential of Decalepis hamiltonii (Wight and Arn). Int J Pharm Pharm Sci 2014;6(6):452-6.

Rami E, Patel I. Quantitative analysis of total phenols and flavonoids in in vivo and in vitro samples of Oroxylum indicum (L.) Vent. Asian J Pharm Clin Res 2015;8(2):202-6.

Kahkonen MP, Hopia AI, Vuorela HJ, Rauha JP, Pihlaja K, Kujala TS, et al. Antioxidant activity of plant extracts containing phenolic compounds. J Agric Food Chem 1999;47:3954-62.

Yang J, Gong ZC, Tan X. Induction of callus and extraction of alkaloid from Yi Mu Cao (Leonurus heterophylus Sw.) culture. Afr J Biotechnol 2008;7(8):1157-62.

Indrani T, Vijay KJ, Sanjeev KA, Maheshwar PT, Birendra P, Upendra KS. Quantitative spectrophotometric estimation of total alkaloids in Withania somnifera L. in vivo and in vitro. Int J Appl Biol Pharm Technol 2016;7(2):254-7.

Babeet ST, Neha S, Soniya C, Rekha V. In vitro preliminary phytochemical and antioxidant activity of Alangium salviifolium linn. Int J Curr Microbiol Appl Sci 2014;3(10):864-72.

Duh PD, Tu YY, Yen GC. Antioxidant activity of water extract of Harng Jyur (Chrysanthemum moifolium Ramat). Lebensm Wiss Technol 1999;32(5):269-77.

Gordon MH. Food antioxidants. In: Hudson BJ, editor. The Mechanism of the Antioxidant Action in Vitro. London: Elsevier; 1990. p. 1-18.

Gulcin I, Oktay M, Kirecci E, Kufrevioglu OI. Screening of antioxidant and antimicrobial activities of anise (Pimpinella anisum L.) seed extracts. Food Chem 2003;83:371-82.

Deepika A, Vidya P. Comparative analysis of in vitro anti-inflammatory and in vivo and in vitro anti-arthritic activity in methanolic extract of Pluchea lanceolata Oliver and Hiern. Int J Biol Pharm Res 2013;4(9):676-80.

Deepika A, Mukesh M, Neha G, Vidya P. In vitro anti-arthritic activity in methanolic extract of Cocculus hirsutus (L.) Diels. in vivo and in vitro. Int J Pharm Sci Res 2014;5(5):1957-62.

Taylor J, Van Staden J. Anti-inflammatory activity in extracts prepared from callus cultures of Eucomis autumnalis (Mill.) Chitt. J Plant Growth Regul 2001;34(3):331-7.




About this article

Title

RAPID IN VITRO CALLOGENESIS AND PHYTOCHEMICAL SCREENING OF LEAF, STEM AND LEAF CALLUS OF MUSSAENDA FRONDOSA LINN. - A MEDICINAL PLANT.

Keywords

Mussaenda frondosa; antioxidant; anti – inflammatory; in vitro culture; phenolics, flavonoids, alkaloids.

DOI

10.22159/ajpcr.2017.v10i6.17527

Date

01-06-2017

Additional Links

Manuscript Submission

Journal

Asian Journal of Pharmaceutical and Clinical Research
Vol 10 Issue 6 June 2017 Page: 81-86

Print ISSN

0974-2441

Online ISSN

2455-3891

Statistics

103 Views | 97 Downloads

Authors & Affiliations

Manasa Dj
Department of Applied Botany, Mangalore University, Mangalagangotri 574199, India
India

Chandrashekar Kr
Department of Applied Botany, Mangalore University, Mangalagangothri, Mangalore - 574 199, Karnataka, India.
India

Bhagya N
Department of Applied Botany, Mangalore University, Mangalagangothri, Mangalore - 574 199, Karnataka, India
India


Article Tools


Email this article (Login required)
Email the author (Login required)

Refbacks

  • There are currently no refbacks.