Articles
PEPPERMINT (MENTHA PIPERITA) SANITATION AND MASS MICROPROPAGATION IN VITRO
Article number
853_8
Pages
77 – 82
Language
English
Abstract
Five plants from each of the 3 peppermint (Mentha piperita) cultivars were kept for one month in a glasshouse under 75% shade.
Subsequently plants were taken to the laboratory and 10 apical growing shoots 5 cm long were removed from each plant and sterilised by soaking in 20% commercial bleach solution with a few drops of Tween 20 for 10 min and washed 3 times in distil sterile water.
In a sterile air laminar flow cabinet, meristems tips 1 to 2 mm long were isolated and cultured in vitro on a modified MS medium and micro-plants produced.
The medium contained MS basic salts and vitamins, supplemented with 3% sucrose, 4.5 mg/L BAP, 0.009 mg/L IBA and 55.7 mg/L ascorbic acid and solidified with 2.5 g/L phytagel.
Cultures incubated in a growth room for two weeks in the dark and constant temperature 21±2°C followed by transfer of cultures to a growth room 16 hours light μmol m-2 s-1 provided by a white and red light fluorescent lambs (50−50%) and 8 hours dark at constant temperature 21±2°C. Proliferation was on an average 4-fold per 2 weeks and rooted in vitro, in the same medium, micro plants produce.
In vitro plant material tested with the molecular method ds-RNA and RT-PCR for viruses and viroids and by ELISA for viruses and other pathogens.
All infected cultures were discarded.
Micro plants were undergoing two faces of hardening the first in vitro by increasing the light intensity to 40 μmol m-2 s-1 for two weeks and the second ex vitro 81 μmol m-2 s-1 provided by AGRO-T-PLUS and in a micro fog were relative humidity (RH) was progressively reduced from 95% to 55%. Plantlets were also hardened in growth chambers with light intensity 40 μmol m-2 s-1 16 hours a day/8 hours dark and constant temperature 21±2°C. This method is use for production of pathogen free and genetically uniform peppermint micro plants to be use as propagation stocks and it can be used if cost permits for the mass micropropagation of new cultivars for rapid release to growers.
Subsequently plants were taken to the laboratory and 10 apical growing shoots 5 cm long were removed from each plant and sterilised by soaking in 20% commercial bleach solution with a few drops of Tween 20 for 10 min and washed 3 times in distil sterile water.
In a sterile air laminar flow cabinet, meristems tips 1 to 2 mm long were isolated and cultured in vitro on a modified MS medium and micro-plants produced.
The medium contained MS basic salts and vitamins, supplemented with 3% sucrose, 4.5 mg/L BAP, 0.009 mg/L IBA and 55.7 mg/L ascorbic acid and solidified with 2.5 g/L phytagel.
Cultures incubated in a growth room for two weeks in the dark and constant temperature 21±2°C followed by transfer of cultures to a growth room 16 hours light μmol m-2 s-1 provided by a white and red light fluorescent lambs (50−50%) and 8 hours dark at constant temperature 21±2°C. Proliferation was on an average 4-fold per 2 weeks and rooted in vitro, in the same medium, micro plants produce.
In vitro plant material tested with the molecular method ds-RNA and RT-PCR for viruses and viroids and by ELISA for viruses and other pathogens.
All infected cultures were discarded.
Micro plants were undergoing two faces of hardening the first in vitro by increasing the light intensity to 40 μmol m-2 s-1 for two weeks and the second ex vitro 81 μmol m-2 s-1 provided by AGRO-T-PLUS and in a micro fog were relative humidity (RH) was progressively reduced from 95% to 55%. Plantlets were also hardened in growth chambers with light intensity 40 μmol m-2 s-1 16 hours a day/8 hours dark and constant temperature 21±2°C. This method is use for production of pathogen free and genetically uniform peppermint micro plants to be use as propagation stocks and it can be used if cost permits for the mass micropropagation of new cultivars for rapid release to growers.
Authors
G.J. Minas
Keywords
propagation, meristem tip culture, peppermint, MS, disease-free
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