Invitro culture of mentha piperita

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  • Stored in Brown bottle at 4oC and pH 5.8
  • Invitro culture of mentha piperita

    1. 1. INVITRO CULTURE OF MENTHA PIPERITA L.THROUGH SHOOT TIP AND SINGLE NODECULTURERoshani Rajbanshi
    2. 2. 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
    3. 3. INTRODUCTION OF NEPAL
    4. 4. INTRODUCTIONArid Tibetan highland inthe north (towards China)Gangetic plains in the south(towards India)Elevation 75m-8,848mClimate Subtropical to Arctic
    5. 5. 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)
    6. 6. 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).
    7. 7. 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.
    8. 8. 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.
    9. 9. 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.
    10. 10. 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
    11. 11. 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.
    12. 12. 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
    13. 13. 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
    14. 14. RESULT Subculture RMS= Rooted Multiple Shoot MS= Multiple ShootNAA(ppm)BAP(ppm)O ppmNode0 ppmShootTip1ppmNode1ppmShootTip2 ppmNode2 ppmShoot Tip0 2/2-RMS2/2-Plantlet2/2-Plantlet2/2RMS2/2-MS 1/2-RMS1/2-MS1 1/2-Plantlet1/2-Plantlet1/2 –RMS1/2 –MS 2/2-MS 1/2-Callus1/2-Plantlet
    15. 15. CALLUS FORMATIONCallus obtained after 4 weeks from Shoot tip culture
    16. 16. 8 WEEKS OLD PLANTLET FROM SHOOT TIPCULTURE Plantlet
    17. 17. 4 WEEKS OLD MULTIPLE SHOOT FROM NODECULTURE
    18. 18. VIGOROUS GROWTH OF MULTIPLE SHOOT AFTER 8WEEKS OF SHOOT TIP IN MS+BAP+NAA
    19. 19. 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.
    20. 20. ACKNOWLEDGEMENT Mukti Ram Aryal Sanjeev Kumar Rai Shankar Pahari
    21. 21. REFERENCES Bentham, G. & Hooker, J.D. (1973). ―GeneraPlantarum” Vol II. Reeve & Co. London. Anonymus. (2000). National Register of MedicinalPlants. HMG Nepal/ IUCN
    22. 22. THANK YOU

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