Articles
IN VITRO PROPAGATION OF AKAMA TULIP VIA ADVENTITIOUS ORGANOGENESIS FROM BULB SLICES
Article number
755_40
Pages
313 – 316
Language
English
Abstract
Five bulbs of Akama tulip (Tulipa cipria akamantis) were sliced 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, slices were placed on the scratched upper surface of a modified MS (Murashige and Skoog, 1962) medium and micro-embryonic shoots were produced.
The medium contained MS basic salts and vitamins, supplemented with 3% sucrose, 4.5 mg L-1 BAP, 0.009 mg L-1 IBA, and 55.7 mg L-1 ascorbic acid and solidified with 2.5 g L-1 phytagel.
Cultures incubated in a growth room for two weeks in the dark and constant temperature of 15±2°C followed by transfer of cultures to a growth room with 16 hours light of 600 lux provided by white and red light fluorescent lambs (50-50%) and 8 hours dark at constant temperature of 15±2°C. Proliferation was on average 4-fold per 2 weeks and rooted in vitro, in the same medium, micro-plants were produced.
In vitro plant material was 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 1500 lux for two weeks and the second ex vitro with light of 1500 lux provided by AGRO-T-PLUS and in a micro-fog where relative humidity (RH) was progressively reduced from 95 to 55%. Plantlets were also hardened in growth chambers with 16 hours a day light intensity of 1500 lux/8 hours dark and constant temperature of 15±2°C. This method may be used for production of pathogen-free and genetically uniform tulip micro-bulbs and micro-plants of Akama tulip bulbs.
In a sterile air laminar flow cabinet, slices were placed on the scratched upper surface of a modified MS (Murashige and Skoog, 1962) medium and micro-embryonic shoots were produced.
The medium contained MS basic salts and vitamins, supplemented with 3% sucrose, 4.5 mg L-1 BAP, 0.009 mg L-1 IBA, and 55.7 mg L-1 ascorbic acid and solidified with 2.5 g L-1 phytagel.
Cultures incubated in a growth room for two weeks in the dark and constant temperature of 15±2°C followed by transfer of cultures to a growth room with 16 hours light of 600 lux provided by white and red light fluorescent lambs (50-50%) and 8 hours dark at constant temperature of 15±2°C. Proliferation was on average 4-fold per 2 weeks and rooted in vitro, in the same medium, micro-plants were produced.
In vitro plant material was 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 1500 lux for two weeks and the second ex vitro with light of 1500 lux provided by AGRO-T-PLUS and in a micro-fog where relative humidity (RH) was progressively reduced from 95 to 55%. Plantlets were also hardened in growth chambers with 16 hours a day light intensity of 1500 lux/8 hours dark and constant temperature of 15±2°C. This method may be used for production of pathogen-free and genetically uniform tulip micro-bulbs and micro-plants of Akama tulip bulbs.
Authors
G.J. Minas
Keywords
Tulipa cipria, in vitro, tulip bulbils production
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