Invitro culture of mentha piperitaPresentation Transcript
INVITRO CULTURE OF MENTHA PIPERITA L.THROUGH SHOOT TIP AND SINGLE NODECULTURERoshani Rajbanshi
OUTLINE Geographical location of Nepal Vegetation of Nepal Introduction of Mentha piperita L. Uses of Mentha piperita L. Objectives Methods Material Result Conclusion Acknowledgement References
INTRODUCTION OF NEPAL
INTRODUCTIONArid Tibetan highland inthe north (towards China)Gangetic plains in the south(towards India)Elevation 75m-8,848mClimate Subtropical to Arctic
VEGETATION OF NEPAL 6500 species flowering plants 4% of flowering plants Endemic to country 4000 species non-flowering plants 450 spp. Pteridophytes 853 spp. Bryophytes 460 spp. Lichens 687 spp. Algae 1600 spp. Fungi(Source: Bentham & Hooker (1973)
MEDICINAL PLANTS OF NEPAL 700 species are recorded as medicinal plants(Chopra et.al. 1885). In 1999, Tiwari reported 1463 species of medicinalplants in Nepal. 50 items of crude herbs and aromatic plants aresold to India and China (HMG, 2000).
INTRODUCTION OF MENTHA L. Nepali name is ―Pudina‖ Found in northern hemisphere Flowering plant Family Labiatae Herbaceous, aromatic and medicinal plant Mentha L. 40 species. (HMG, 2000) M. piperita hybrid of M. spicata and M. aquatica.
USES Peppermint Aromatic, stimulant Used to treat stomachache, allaying nausea,flatulence and vomiting. A good source of peppermint oil Peppermint oil is used in pharmaceuticals, dentalpreparation, mouthwashes, cough drops, soaps,chewing gums and candies. Also used in ―Chatney‖ in South Asia.
OBJECTIVES To establish in-vitro culture in different hormoneconcentration. To observe the response in different hormoneconcentration. To find the variation in single node culture andshoot tip culture.
METHODOLOGY Murashinge and Skoog (1962) medium was preparedwith the following nutrients Macro nutrients MgSO4.7H2O, KH2PO4, KNO3,NH4NO3, CaCl2.2H2O Micro nutrients H3BO3, MnSO4.4H2O,ZnSO4.7H2O, NaMoO4.2H2O, CuSO4.5H2O,COCl2.6H2O, KI Iron Source FeSO4.7H2O, NaEDTA.2H2O Vitamins Thiamin HCl, Pyridoxin HCl, Nicotinicacid, Myoinositol, Glycin Carbon source Sucrose Hormones Naphthalene acetic acid(NAA) andBenzyl amino purine (BAP) 0.8 % Difco-facto agar solidification
MATERIAL—MENTHA PIPERITA L. Sterilized in running water for 1 hour, 1% sodiumhypochloride for 5 minutes, 70% ethanol for 1minute, rinsed in distilled water for 3 times Single nodes and shoot tip of 4-5 mm pieces werecut Explants were inoculated in culture tubes insidelaminar air flow, sealed and transferred toincubation room Temperature +/- 25oC Observation 4-8 weeks Subculture of plantlets were done in differenthormones concentration.
RESULTInvitro Culture of Single Node MS = Multiple Shoot RMS = Rooted Multiple ShootNAA (ppm)BAP (ppm) 0 1 204/4-RMS2/2-SingleShoot 2/2-Single Shoot1 1/4-plantlet3/4-RMS 2/2-MS1/2 -shoot1/2 -poor growth
RESULTInvitro culture of Shoot tip MS = Multiple Shoot RMS = Rooted Multiple ShootNAA(ppm)BAP(ppm)0 1 20 2/2-Plantlet 1/2 – MS 1/2 - MS11/2-plantlet1/2-Callus1/2 –Plantlet 2/2- MS
CALLUS FORMATIONCallus obtained after 4 weeks from Shoot tip culture
8 WEEKS OLD PLANTLET FROM SHOOT TIPCULTURE Plantlet
4 WEEKS OLD MULTIPLE SHOOT FROM NODECULTURE
VIGOROUS GROWTH OF MULTIPLE SHOOT AFTER 8WEEKS OF SHOOT TIP IN MS+BAP+NAA
CONCLUSION Both single node and shoot tip can be culturedartificially. Basal media is the best media to obtain plantlet androoted multiple shoot. In few cases, addition of NAA suppressed theproliferation of multiple shoot. The basal media supplemented with BAP (2ppm)and NAA (1ppm) gave high number of multipleshoot.