Lambardi Maurizio, De Carlo Anna, Capuana Maurizio
Istituto per la Valorizzazione del Legno e delle Specie Arboree, CNR/Consiglio Nazionale delle Ricerche, Polo Scientifico, via Madonna del Piano, 50019 Sesto Fiorentino (Firenze), Italy.
Cryo Letters. 2005 May-Jun;26(3):185-92.
An effective procedure for the cryopreservation of horse chestnut (Aesculus hippocastanum L.) embryogenic callus by vitrification/one-step freezing is described here. In particular, the study focused on the possibility of recovering the full proliferation potential of the embryogenic lines after storage in liquid nitrogen. The developmental stage of the embryogenic lines was shown to play an important role. Ninety-min incubation in PVS2 and preservation at -196 degrees C of callus samples, containing a prevalence of embryogenic masses at an advanced stage of somatic embryo maturation (i.e., the torpedo stage), gave optimum regrowth of healthy and proliferating embryogenic callus. Moreover, raising the thawing temperature to 45 degrees C yielded the maximum survival (94%) of torpedo-stage embryogenic samples, recovery of proliferation and, in more than 70% of cases, maturation to the cotyledonary stage. This study opens the way to the possibility of safe, long-term storage in liquid nitrogen of valuable embryogenic lines of horse chestnut, avoiding repeated subculturing.
本文描述了一种通过玻璃化/一步冷冻法对七叶树（Aesculus hippocastanum L.）胚性愈伤组织进行有效冷冻保存的方法。特别地，该研究聚焦于液氮保存后胚性系恢复其全部增殖潜力的可能性。结果表明胚性系的发育阶段起着重要作用。将处于体细胞胚成熟后期（即鱼雷期）且胚性团块占优势的愈伤组织样品在PVS2中孵育90分钟，并在-196℃保存，可使健康且增殖的胚性愈伤组织实现最佳再生长。此外，将解冻温度提高到45℃可使鱼雷期胚性样品的存活率达到最高（94%），恢复增殖能力，并且在超过70%的情况下可成熟至子叶期。这项研究为七叶树珍贵胚性系在液氮中进行安全、长期保存开辟了道路，避免了反复继代培养。
