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Abstract
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| Cryopreservation is a reliable option for the long-term storage of clonal species. Many mint (Mentha spp.) genotypes are clonally propagated. Cryopreservation protocols that achieve high recovery are available for several Mentha genotypes using several techniques: controlled cooling, vitrification, encapsulation-vitrification, encapsulation-dehydration, and droplet-vitrification. Not all mint genotypes respond similarly to different cryopreservation techniques. Besides, small differences in the procedures carried out, in what they could seem similar protocols, could account for different survival rates. The droplet-vitrification procedure used by Senula et al. (2007) varies to other droplet methods in the fact that the aluminium foil strip (containing the apices immersed in PVS2 droplets) is included in a cryovial before its immersion in LN, while in other procedures the strips are directly immersed in the LN. We compared in three mint genotypes these two procedures combined with desiccation in PVS2 for 20 min at room temperature or for 30 min at 0ºC. In the first case (cryovial immersed in LN) warming of cryovials was performed by immersing them in a 40ºC water bath for 3-5 seconds (followed by immersion in washing solution, 1.2 M sucrose, for 20 min at room temperature); and in the second case, by direct immersion of the foil strips in the washing solution at room temperature. Similar results (recovery) among treatments were obtained in all genotypes. The influence of different preculture procedures on survival after recovery from liquid nitrogen was evaluated with two genotypes, using the standard droplet technique (direct immersion in LN). Different combinations of nodal segment preculture (3 days at 25ºC, 3 weeks at 10ºC and 2 weeks at -1º/25ºC) and shoot apex preculture (5º or 25ºC on MS or MS+ 0.3 M sucrose) were studied. Apices were previously desiccated with PVS2 for 30 min at 0ºC. Best results were obtained with -1º/25C¿ 25ºC MS+ 0.3 M sucrose in one genotype, and 25ºC¿25ºC MS+ 0.3 M sucrose, in the other one. | |
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International
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Si |
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Congress
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47th annual meeting of Society for Cryobiology |
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960 |
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Place
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Bristol (Reino Unido) |
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Reviewers
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Si |
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ISBN/ISSN
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0011-2240 |
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Start Date
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17/07/2010 |
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End Date
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20/07/2010 |
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From page
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1 |
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To page
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1 |
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Cryopreservation of mint in vitro apices: effect of pre-culture and cooling procedures |