Cryopreservation of cold-acclimated mint (Mentha spp.) shoot tips using a simple vitrification protocol.
Accessions of Mentha x piperita, M. x villosa, and M. spicata were evaluated for regrowth after cooling in liquid nitrogen using shoot tips from in-vitro grown plantlets and a simple vitrification protocol with aluminium foil as a carrier. The influences of plant preculture, loading solution and loading time and of the effects of the cryoprotectant PVS 2 on plant re-growth after re-warming were investigated. Nodal segments were cultivated at constant temperatures of 20 or 25 degree C or in alternating temperature regimes (25/15C or 25/-1C). The illumination was always 16 h per day. The re-growth levels after re-warming were significantly higher in plants pre-cultured at 25/-1C regime than in plants cultivated at 20C or 25C or at 25/15C regime for all nine tested accessions. The mean re-growth levels increased from 36 percent at 20C to 69percent at alternating temperatures, respectively. The maximum of plant re-growth after re-warming was 89 percent. A pre-culture at alternating temperatures of 25/15C did not increase the recovery of plants. Loading in sucrose solutions with different dehydration capacities did not alter the plant re-growth. Differences in the loading time between 20 min and 2 h were not important for re-growth either. No significant differences were found between freezing without and with PVS 2 droplets on the aluminium foil. Re-grown shoots rooted easily on the re-growth medium and plantlets were successfully transferred to soil.
Senula
Angelika
A
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany.
Joachim Keller
E R
ER
Sanduijav
Tekshbayar
T
Yohannes
Tamene
T
D000064:Acclimatization; D000535:Aluminum; D018070:Biological Specimen Banks; D003080:Cold Temperature; D015925:Cryopreservation / Q000379:methods*; D046508:Culture Techniques; D036142:Mentha piperita / Q000254:growth & development*; D018520:Plant Shoots / Q000254:growth & development; D012987:Soil; D012996:Solutions; D014867:Water
D012987:Soil; D012996:Solutions; D014867:Water; D000535:Aluminum