Invitro culture of mentha piperita
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INVITRO CULTURE OF MENTHA PIPERITA L. THROUGH SHOOT TIP AND SINGLE NODE CULTURE
Roshani Rajbanshi
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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
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INTRODUCTION OF NEPAL
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INTRODUCTION
Arid Tibetan highland in the north (towards China)
Gangetic plains in the south (towards India)
Elevation 75m-8,848m Climate Subtropical to
Arctic
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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)
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MEDICINAL PLANTS OF NEPAL
700 species are recorded as medicinal plants (Chopra et.al. 1885).
In 1999, Tiwari reported 1463 species of medicinal plants in Nepal.
50 items of crude herbs and aromatic plants are sold to India and China (HMG, 2000).
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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.
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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,
dental preparation, mouthwashes, cough drops, soaps, chewing gums and candies.
Also used in “Chatney” in South Asia.
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OBJECTIVES
To establish in-vitro culture in different hormone concentration.
To observe the response in different hormone concentration.
To find the variation in single node culture and shoot tip culture.
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METHODOLOGY Murashinge and Skoog (1962) medium was prepared
with 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, Nicotinic
acid, Myoinositol, Glycin Carbon source Sucrose Hormones Naphthalene acetic acid(NAA) and Benzyl
amino purine (BAP) 0.8 % Difco-facto agar solidification
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MATERIAL—MENTHA PIPERITA L.
Sterilized in running water for 1 hour, 1% sodium hypochloride for 5 minutes, 70% ethanol for 1 minute, rinsed in distilled water for 3 times
Single nodes and shoot tip of 4-5 mm pieces were cut
Explants were inoculated in culture tubes inside laminar air flow, sealed and transferred to incubation room
Temperature +/- 25oC Observation 4-8 weeks Subculture of plantlets were done in different
hormones concentration.
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RESULT
Invitro Culture of Single Node
MS = Multiple Shoot RMS = Rooted Multiple Shoot
NAA (ppm)\BAP (ppm) 0 1 2
04/4-RMS
2/2-Single Shoot 2/2-Single Shoot
11/4-plantlet 3/4-RMS 2/2-MS
1/2 -shoot 1/2 -poor growth
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RESULT
Invitro culture of Shoot tip
MS = Multiple Shoot RMS = Rooted Multiple Shoot
NAA (ppm)\BAP (ppm)
0 1 2
0 2/2-Plantlet 1/2 – MS 1/2 - MS1
1/2-plantlet1/2-Callus1/2 –Plantlet 2/2- MS
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RESULT
Subculture
RMS= Rooted Multiple Shoot MS= Multiple Shoot
NAA (ppm)\BAP (ppm)
O ppmNode
0 ppmShoot Tip
1ppmNode
1ppmShoot Tip
2 ppmNode
2 ppmShoot Tip
0 2/2-RMS
2/2-Plantlet
2/2-Plantlet
2/2 RMS
2/2-MS 1/2-RMS1/2-MS
1 1/2-Plantlet
1/2-Plantlet
1/2 – RMS
1/2 –MS
2/2-MS 1/2-Callus1/2-Plantlet
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CALLUS FORMATION
Callus obtained after 4 weeks from Shoot tip culture
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8 WEEKS OLD PLANTLET FROM SHOOT TIP CULTURE
Plantlet
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4 WEEKS OLD MULTIPLE SHOOT FROM NODE CULTURE
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VIGOROUS GROWTH OF MULTIPLE SHOOT AFTER 8 WEEKS OF SHOOT TIP IN MS+BAP+NAA
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CONCLUSION
Both single node and shoot tip can be cultured artificially.
Basal media is the best media to obtain plantlet and rooted multiple shoot.
In few cases, addition of NAA suppressed the proliferation of multiple shoot.
The basal media supplemented with BAP (2ppm) and NAA (1ppm) gave high number of multiple shoot.
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ACKNOWLEDGEMENT
Mukti Ram Aryal Sanjeev Kumar Rai Shankar Pahari
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REFERENCES
Bentham, G. & Hooker, J.D. (1973). “Genera Plantarum” Vol II. Reeve & Co. London.
Anonymus. (2000). National Register of Medicinal Plants. HMG Nepal/ IUCN
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THANK YOU