A cryopreservation protocol for immature zygotic embryos of species of Ilex (Aquifoliaceae)
Biocell
;
32(1): 33-39, Apr. 2008. ilus, tab
Article
Dans Anglais
| LILACS
| ID: lil-541120
ABSTRACT
Tropical Ilex species have recalcitrant seeds. This work describes experiments demonstrating the feasibility of long-term conservation of Ilex brasiliensis, I. brevicuspis, I. dumosa, I. intergerrima, I. paraguariensis, I. pseudoboxus, I. taubertiana, and I. theezans through cryopreservation of zygotic rudimentary embryos at the heart developmental stage. The embryos were aseptically removed from the seeds and precultured (7 days) in the dark, at 27 +/- 2 degrees C on solidified (0.8% agar) 1/4MS medium, [consisting of quarter-strength salts and vitamins of Murashige and Skoog (1962) medium] with 3% sucrose and 0.1 mg/l Zeatin.The embryos were then encapsulated in 3% calcium alginate beads and pretreated at 24 h intervals in liquid medium supplemented with progressively increasing sucrose concentrations (0.5, 0.7 5 and 1 M). Beads were dehydrated for 5 h with silicagel to 25% water content (fresh weight basis) and then placed in sterile 5 ml cryovials. Then the beads were either plunged rapidly in liquid nitrogen were they were kept for 1 h (rapid cooling) or cooled at 1 degrees C min(-1) to -30 degrees C. Then the beads were immersed in liquid nitrogen for 1 h (slow cooling). The beads were rewarmed by immersion of the cryovials for 1 min in a water bath thermostated at 30 degrees C. Finally, beads were transferred onto culture medium (1/4MS, 3% sucrose, 0.1 mg/l zeatin, solidified with 0.8% agar) and incubated in a growth room at 27 +/- 2 degrees C under a 14 h light (116 micromol. m(-2) x s(-1))/ 10 h dark photoperiod. Maximum recovery percentages between 15 and 83% (depending on de the species and the treatment) were obtained with the cryopreserved embryos.
Texte intégral:
Disponible
Indice:
LILAS (Amériques)
Sujet Principal:
Graines
/
Cryoconservation
/
Survie cellulaire
/
Ilex
Type d'étude:
Études d'évaluation
langue:
Anglais
Texte intégral:
Biocell
Thème du journal:
Clulas
Année:
2008
Type:
Article
Pays d'affiliation:
Argentine
Institution/Pays d'affiliation:
Uiversidad Nacional del NordEste/AR
/
Universidad Nacional del NordEste/AR
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