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Invitro culture of mentha piperita
1. INVITRO CULTURE OF MENTHA PIPERITA L.
THROUGH SHOOT TIP AND SINGLE NODE
CULTURE
Roshani Rajbanshi
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
4. INTRODUCTION
Arid Tibetan highland in
the north (towards China)
Gangetic plains in the south
(towards India)
Elevation 75m-8,848m
Climate Subtropical to Arctic
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. 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).
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. 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.
9. 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.
10. 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
11. 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.
12. RESULT
Invitro Culture of Single Node
MS = Multiple Shoot
RMS = Rooted Multiple Shoot
NAA (ppm)BAP (ppm) 0 1 2
0
4/4-RMS
2/2-Single
Shoot 2/2-Single Shoot
1 1/4-plantlet
3/4-RMS 2/2-MS
1/2 -shoot
1/2 -poor growth
13. 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 - MS
1
1/2-plantlet
1/2-Callus
1/2 –Plantlet 2/2- MS
18. VIGOROUS GROWTH OF MULTIPLE SHOOT AFTER 8
WEEKS OF SHOOT TIP IN MS+BAP+NAA
19. 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.
