Beule T, Ilbert P, Adeoti K, Durand-Gasselin T, Dumet D, Engelmann F, Morcillo F
CIRAD, Universite Montpellier, UMR DIADE, F-34398 Montpellier, France.
Universite d'Abomey-Calavi (UAC), Faculte des Sciences et Techniques (FAST), Departement de Biologie Vegetale, Laboratoire de Microbiologie et des Technologies Alimentaires (LAMITA), Cotonou, Benin.
Cryo Letters. 2018 Jan/Feb;39(1):60-66.
BACKGROUND: A cryopreservation protocol has been established for oil palm somatic embryos (SEs), the efficiency of which must be evaluated, both in terms of regeneration and of long-term storage capacity, before its large-scale routine use.
To test the survival and recovery of 29 clones of oil palm somatic embryos cryostored for 20 years.
Clumps of SEs were pregrown for 7 days on medium containing 0.75 M sucrose, dehydrated in air-tight containers containing silica gel to moisture contents between 19-35% fresh weight, and then immersed directly in liquid nitrogen and stored in cryotanks for 20 years.
Survival of SEs cryopreserved and rewarmed immediately displayed an average value of 19.1% for the 29 clones tested while survival of SEs rewarmed after 20 years of cryostorage was significantly higher, with an average of 33.2% for the 28 surviving clones. Out of these 28 surviving clones, three were lost due to contamination or regrowth decline, six produced only shoots and the rest proliferated.
It is possible to cryostore oil palm SEs for extended periods and to regenerate proliferating cultures and plantlets from the cryopreserved material. The cryopreservation protocol established can thus be efficiently used to store oil palm germplasm and to manage large-scale production in industrial laboratories.
背景:已建立油棕体细胞胚(SEs)的冷冻保存方案,在其大规模常规使用之前,必须从再生和长期储存能力方面评估其效率。
测试冷冻保存20年的29个油棕体细胞胚克隆的存活率和恢复情况。
将体细胞胚团块在含有0.75M蔗糖的培养基上预培养7天,在装有硅胶的气密容器中脱水至水分含量为鲜重的19 - 35%,然后直接浸入液氮并在低温罐中储存20年。
对于测试的29个克隆,立即冷冻保存并复温的体细胞胚平均存活率为19.1%,而冷冻保存20年后复温的体细胞胚存活率显著更高,28个存活克隆的平均存活率为33.2%。在这28个存活克隆中,3个因污染或生长衰退而丢失,6个仅产生芽,其余的进行了增殖。
油棕体细胞胚可以长期冷冻保存,并能从冷冻保存的材料中再生增殖培养物和植株。因此,所建立的冷冻保存方案可有效地用于储存油棕种质并在工业实验室中管理大规模生产。