Cryopreservation of shoot tips from dormant buds of Diospyros kaki by one-step vitrification (Articolo in rivista)

Type
Label
  • Cryopreservation of shoot tips from dormant buds of Diospyros kaki by one-step vitrification (Articolo in rivista) (literal)
Anno
  • 2009-01-01T00:00:00+01:00 (literal)
Alternative label
  • Carla Benelli ; E. Giordani ; A. De Carlo ; S. Pecchioli (2009)
    Cryopreservation of shoot tips from dormant buds of Diospyros kaki by one-step vitrification
    in Cryo-letters
    (literal)
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  • Carla Benelli ; E. Giordani ; A. De Carlo ; S. Pecchioli (literal)
Pagina inizio
  • 394 (literal)
Pagina fine
  • 395 (literal)
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  • 30 (literal)
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  • 2 (literal)
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  • 5 (literal)
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  • ISI Web of Science (WOS) (literal)
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  • IVALSA/Istituto per la Valorizzazione del Legno e delle Specie Arboree, National Research Council (CNR), Sesto Fiorentino (Firenze), ITALY ; Department of Horticulture, University of Florence, Sesto Fiorentino (FI), Italy ; IVALSA/Istituto per la Valorizzazione del Legno e delle Specie Arboree, National Research Council (CNR), Sesto Fiorentino (Firenze), ITALY ; Department of Horticulture, University of Florence, Sesto Fiorentino (FI), Italy (literal)
Titolo
  • Cryopreservation of shoot tips from dormant buds of Diospyros kaki by one-step vitrification (literal)
Abstract
  • The safeguard genetic resources of persimmon in the Mediterranean basin is limited. Some European institutions have contributed, through EC GENRES29 Project 'Conservation of minor fruit tree species', to the safeguarding and conservation of a number of persimmon accessions collected in clonal orchards. Cryopreservation could be an advanced technique to reduce serious implications and risks in the management of germplasm collections and preserve Diospyros kaki cultivars. To optimize cryopreservation procedure for shoot tips from dormant buds of persimmon, mainly used in the Mediterranean area, several experiments were carried out by one-step vitrification method. Vegetative dormant buds were taken, during the winter, from twigs of persimmon cultivars ('Kaki Tipo', 'Jiro', 'Triumph', 'Hiratanenashi' and 'Rojo Brillante'). Shoot tips (2-3 mm) were dissected from sterilized buds and precultured on MS medium (modified with half-strength NH4NO3 and KNO3) with 0.3 M sucrose for 48h. To induce osmodehydration, the shoot tips were loaded in Plant Vitrification Solution 2 (PVS2), for different times (20 min in the first experiment; 30, 60, 90 min in the following experiments) and two treatment temperatures (0°C and 25°C). The explants were rapidly frozen to -196°C by direct immersion in liquid nitrogen (LN). For recovery, they were quickly rewarmed in a waterbath at 40°C for 1 min, then the shoot tips were transferred on the regrowth medium (MS modified with 0.06 M sucrose, 22.2 mM BA, 2 g/l gerlite) and maintained at 24°C with 16h photoperiod. Survival of cryopreserved shoot tips was assessed after 3 weeks. By applying a treatment with PVS2 for 20 min at 25°C on shoots tips from dormant buds, the survival rate observed in cv 'Triumph' (25%), 3 weeks after cryopreservation, is lower than those reported by Matsumoto et al. (2001). In a subsequent experiment on cv 'Kaki Tipo', where we applied a longer exposure time to PVS2, a higher shoot tip survival rate was obtained, in particular with PVS2 treatment for 90 min (73%). The same protocol applied on other cultivars confirmed the effectiveness of this vitrification procedure and with 90 min PVS2 treatment, the survival percentage in three cultivars ('Jiro', 'Triumph' and 'Rojo Brillante') was recorded between 56-80%. Whereas, in cv 'Hiratanenashi', the best survival (86%) was observed with 60 min PVS2 treatment before explant immersion in LN. Molecular assays were performed with fresh material, in vitro-cultured and cryopreserved shoots tips of three persimmon cultivars ('Kaki Tipo', 'Jiro', 'Triumph') in order to validate both the micropropagation procedure and the optimized cryopreservation technique. In our experiments, no morphological deviation was observed in the development of persimmon cryopreserved plantlets. (literal)
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