Tissue Culture Studies of Celastrus Paniculatus and Evaluation of Secondary Metabolites

“Tissue Culture Studies of Celastrus paniculatus and Evaluation of secondary metabolites”

SYNOPSIS FOR

M.PHARM DISSERTATION

SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA.

BY

ANANTH.A

DEPARTMENT OF PHARMACOGNOSY

P E S COLLEGE OF PHARMACY,

BENGALURU-560050, KARNATAKA.

(2010-2011)

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE.

ANNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR

DISSERTATION.

1. / Name of the candidate and Address: / ANANTH.A
P.E.S COLLEGE OF PHARMACY
50FT.ROAD,
HANUMANTHNAGAR
BENGALURU-560050
PERMANENT ADDRESS
MAHADI SUBBAMMAS HOUSE,
PALEGAR STREET , ANEKAL
BENGALURU-562106
2. / Name of the Institution / P.E.S COLLEGE OF PHARMACY
50FT.ROAD,
HANUMANTHNAGAR,
BENGALURU-560050.
3. / Course of study and subject: / MASTER OF PHARMACY IN PHARMACOGNOSY.
4. / Date of admission to the course / 16TH JUNE 2009
5. / Title of the topic:
“TISSUE CULTURE STUDIES OF CELASTRUS PANICULATUS (CELASTRACEAE) AND EVALUATION OF SECONDARY METABOLITES”
6. / Brief resume of the intended work:
6.1: Need to the study:
Ø  Celastrus paniculatus (Malkangani) a large climbing unarmed shrub with long slender elongating branches which are reddish brown and covered with elongate white lenticles belonging to the family Celastraceae. Leaves are simple, alternate, ovate and glabrous. Fruit capsules, depressed globose, three lobed, bright yellow when ripe. Seeds are completely enclosed in an orange red aril.
Ø  This plant is widely used in traditional system of medicine and it is reported to possess thermogenic, digestive, laxative, emetic, emmenagouge, aphrodisiac, anti-inflammatory, cardiotonic, diuretic, abortificient, rheumatism, gout, brain tonic. This plant is having the property of stimulating the intellect and sharpening the memory and the rate of growth of the plant by conventional method of cultivation is slow, hence tissue culture studies on this plant will be helpful to preserve the plant for use in future.1
Ø  It contains sesquiterpene alkaloids such as celapagine, celapanigine, celapanine, celatrol. and some other substances such as linoleic, linolenic, palmetic, stearic acid, polyhydric alcohol-malkanguniol along with β-amyrine and β-sitosterol. Hence the present work will be important for additional research input regarding the production of this plant by tissue culture.1-2
Ø  The process of micropropagation uses various phytohormones in different concentration. Establishing a right proportion of these hormones for a particular variety will result in the production of large number of plantlets in short period of time.
Ø  Micropropagation is a unique method for production of uniform and disease free plants. This technique would facilitate to obtain large number of uniform plants irrespective of season and will serve as an alternative source of seed material. In-vitro conservation of germplasm is a safe method to protect species & reducing the risk of natural vagaries.
Ø  Generic nature of plant plays a major role in defining the quantity of active principles. Cultivation and production of secondary metabolites is constant under standard conditions. Hence production of plant by tissue culture method will be helpful in producing the plants of specific type of significant importance.
Ø  Estimation of secondary metabolite content in in-vitro regenerated plant by tissue culture and comparing it with the content of naturally propagated plant has not been yet reported. Thus the present work is an attempt to know whether the secondary metabolites have increased in in-vitro grown plant or not.
Ø  Considering the importance of above parameters, it is concluded that research work on tissue culture studies of Celastrus paniculatus plant is of significance.
6.2: Review of literature.
Ø  Martin et.al., (2006) established an efficient micropropagation system for Celastrus paniculatus willd. and observed qualitative chemical similarity of the tissue culture regenerants with the mother plant using HPTLC.3
Ø  Sharada.M et.al., (2002) regenerated plantlet of Celastrus paniculatus willd. from cotyledonary leaf derived callus using MS medium supplement with NAA and Kinetin.4
Ø  Nair L.G and Seeni.S (2001) cultured nodes, shoot tips, internodes and leaf bases of Celastrus paniculatus in MS medium containing agar, sucrose and benzylaminopurine and kinetin. The plants generated were uniform in morphology, growth and cytological characteristics.5
Ø  P.B Godkar et.al., (2003) have reported attenuation of hydrogen peroxide and glutamate induced injury in embryonic rat forebrain neuronal cells by seed oil and organic extract of Celastrus paniculatus.6
Ø  M.H.V Kumar and Y.K Gupta (2002) have reported antioxidant property of Celastrus paniculatus.7
Ø  K Nalini et.al (1995) have reported effects of Celastrus paniculatus of passive avoidance performance biogenic amine turnover in albino rats.8
Ø  Ramamoorthy Rajkumar et.al (2007) have evaluated the anxiolytic potential of Celastrus oil extracted from Celastrus paniculatus in rat models of behavior.9
Ø  Praful B Godkar et.al., (2004) have reported neuroprotective effects of Celastrus paniculatus – seed water soluble extract on glutamate induced toxicity.10
Ø  Mohamed Fawzy Ramadan et.al., (2009) has reported radical scavenging activity of Celastrus paniculatus seed oil.11
Ø  Two new β-dihydroagarofuran sesquiterpene polyesters, 1-eq,6-eq,8-eq,12-tetraacetoxy-
9-eq-benzoxy-β-dihydroagarofuran and 1-eq,8-eq,12-triacetoxy-9-ax-furancarboxy-β-
dihydroagarofuran, have been isolated from Celastrus paniculatus. Their structures have
been established by means of extensive NMR studies.12
Ø  Two new β-dihydroagarofuran sesquiterpene polyol esters were isolated from Celastrus paniculatus. Their structures were deduced on the basis of spectral analysis including 2D NMR.13
Ø  N.L. Raju and M.N.V. Prasad carried out the effect of growth hormone on adventitious
root formation in semi-hardwood cuttings of Celastrus paniculatus.14
Ø  M.S. Rao and S.D.Prohit have reported the effect of different growth regulators on induction of multiple shoot buds directly from the internode explants of Celastrus paniculatus.15
7. / 6.3: Main objectives of the study:
Ø  Plant regeneration of Celastrus paniculatus by tissue culture method.
Ø  To study the effect of hormones on tissue culture of Celastrus paniculatus.
Ø  Study and quantify the secondary metabolites from the in- vitro regenerated plants using HPTLC and GC-MS and comparing it with the constituents of naturally propagated plant.
Materials and methods:
7.1: Source of the data:
The required data will be obtained from:
Ø  Electronic data (Internet).
Ø  Review articles from journals.
Ø  Journals searched on RGUHS digital library.
Ø  Library of IISC, PES College of Pharmacy and IIHR
Place of work: Division of plant genetic resource, IIHR, Bengaluru.
PES College of pharmacy, Bengaluru.
7.2 Methods of collection of the data (including sampling procedure if any)
1.  Collection and authentication of the plant material
2.  Data, regarding the optimum requirements of hormones required for the in vitro multiplication of plants.
3.  Studies regarding the quantity of secondary metabolites in plant regenerated through micro propagation method and comparing it with naturally propagated plant.
4.  Quantification of secondary metabolites by HPTLC and GC-MS method.
7.3: Does the study require any investigations or intervention to be conducted on patients other human or animals? If so, please describe briefly:
- No –
7.4: Has ethical clearance been obtained from your institute in case of 7.3
-- Not applicable—
List of references:
1.  Prajapti, Purohit, Sharma. A Hand Book of Medicinal Plants. First ed; 2003.
2.  Joshi SG. Medicinal Plants. : Oxford and IBH; 2003.
3.  Martin G, P. GS, Raja, S. S, Raghu. An efficient micropropagation system for Celastrus paniculatus Willd. a vulnerable medicinal plant. Journal of Forest Research. 2006; 11(5):461-5
4.  Sharada.M, Ahuja.A, Kaul.M.K. Regeneration of plantlets via callus cultures in Celastrus paniculatus willd-A rare endangered medicinal plant. Journal of Plant Biochemistry and Biotechnology (India). 2003; 12(1):65-9.
5.  Nair. L.G, Seeni.S. Rapid in vitro multiplication and restoration of Celastrus paniculatus Willd. sub sp. paniculatus (Celastraceae), a medicinal woody climber. Indian Journal Exp Biol. 2001; 39(7):697-704.
6.  Godkara PB, Gordona RK, Ravindranb A, Doctora BP. Celastrus paniculatus seed oil and organic extracts attenuate hydrogen peroxide- and glutamate-induced injury in embryonic rat forebrain neuronal cells. Phytomedicine. 2003;13:29–36.
7.  Kumar MHV, Gupta YK. Antioxidant Property of Celastrus Paniculatus Willd.: A Possible Mechanism in Enhancing Cognition. Phytomedicine. 2002 9:302-11.
8.  Nalini K, Karanth KS, Rao A, Aroor AR. Effects of Celastrus paniculatus on Passive Avoidance Performance and Biogenic Amine Turnover in Albino Rats. Journal of Ethnopharmacology 1995 47(2):101-8.
9.  Rajkumar R, Kumar EP, Sudha S, Suresh B. Evaluation of Anxiolytic Potential of Celastrus Oil in Rat Models of Behaviour. Fitoterapia. 2007 78(2):120-4.
10.  B P, Godkar, K R, Gordon. Celastrus paniculatus Seed Water Soluble Extracts Protect against Glutamate Toxicity in Neuronal Cultures from Rat Journal of Ethno pharmacology 2004;93:213-9.
11.  Ramadan MF, Kinni SG, Rajanna LN, Seetharam YN. Fatty acids, bioactive lipids and radical scavenging activity of Celastrus paniculatus Willd. Seed oil. Scientia Horticulturae. 2009;123:104–9.
12.  Sang H, Wang H, Tu Y, Chen Y. Dihydroagarofuran sesquiterpenoids from Celastrus paniculatus. Phytochemistry. 1991; 30(5):1547-9.
13.  Zhang K, Wang Y, Chen Y. Sesquiterpenes from Celastrus paniculatus. Phytochemistry. 1998;48(6):1067,9
14.  Raju NL, Prasad MNV. Influence of growth hormones on adventitious root formation in semi-hardwood cuttings of Celastrus paniculatus. Journal Agroforestry Systems. 2009.
15.  M.S.Rao, Purohit SD. In vitro shoot bud differentiation and plantlet regeneration in Celastrus paniculatus. Journal Biologia Plantarum 2006; 50:501-6.
08.
/ NAME OF THE CANDIDATE
/
ANANTH.A
/
09.
/
SIGNATURE OF THE CANDIDATE
/ (ANANTH.A)
/
10.
/ REMARKS OF THE GUIDE
/
11.
/ 11.1 NAME AND DESIGNATION OF
THE GUIDE
/ Dr. K. LAKSHMAN
PROFESSOR AND HEAD
DEPT. OF PHARMACOGNOSY
P.E.S.COLLEGE OF PHARMACY,
BENGALURU-560050.
/
11.2 SIGNATURE
/
11.3 HEAD OF THE DEPARTMENT
/ Dr. K. LAKSHMAN
PROFESSOR AND HEAD
DEPT. OF PHARMACOGNOSY
P.E.S.COLLEGE OF PHARMACY,
BENGALURU-560050
/
11.4 SIGNATURE
/
11.5 CO GUIDE
/ Dr. P.E. RAJASEKHARAN
SENIOR SCIENTIST
DIVISION OF PLANT GENETIC
RESOURCES, IIHR
BENGALURU.
/
12.
/ 12.1 REMARKS OF THE PRINCIPAL
/
/ 12.2 SIGNATURE
/ PROF. Dr. S. MOHAN
PRINCIPAL,
P.E.S COLLEGE OF PHARMACY
BENGALURU-560050.
/