Kaya E, Alves A, Rodrigues L, Jenderek M, Hernandez-Ellis M, Ozudogru A, Ellis D
USDA-ARS National Center for Genetic Resources Preservation, Fort Collins, Colorado, USA. Gebze Institute of Technology, Molecular Biology and Genetics Department, Kocaeli, Turkey.
USDA-ARS National Center for Genetic Resources Preservation; mbrapa Labex-USA, USDA-ARS, Fort Collins, Colorado, USA.
Cryo Letters. 2013 Nov-Dec;34(6):608-18.
The long-term preservation of forest genetic resources is a vital part of preserving our forest crops for future generations. Unfortunately, there are few genebanks dedicated to forest trees and very few methods for long-term preservation of forest genetic resources collections aside from field plantings of a limited number of seed-derived or elite clonal individuals. The use of cryopreservation for the long-term storage of elite germplasm is increasingly being used for the long-term preservation of clonal agronomic crops but for forest trees, such as Eucalyptus, the methodology for cryopreservation of diverse genetic resources collections has not been established. We report the successful cryopreservation of a germplasm collection of in vitro shoot cultures of thirteen Eucalyptus spp. lines consisting of two E. grandis x E. camaldulensis lines, seven E. urophylla x E. grandis lines, one E. grandis line, two E. grandis x E. urophylla lines, and one E. camaldulensis line. In a comparison of two cryopreservation methods, sucrose sensitivity limited the application of encapsulation-dehydration. However, with droplet-vitrification, all thirteen lines had good survival after cryopreservation in liquid nitrogen. A 30 min exposure to Plant Vitrification Solution 2 (PVS2) yielded post-liquid nitrogen survival between 38% and 85% depending on the line. One hundred shoot tips from all thirteen lines are currently in long-term storage as a germplasm collection.
森林遗传资源的长期保存是为子孙后代保护我们的森林作物的重要组成部分。不幸的是，专门用于林木的基因库很少，除了对有限数量的种子衍生或优良无性系个体进行田间种植外，几乎没有长期保存森林遗传资源收集品的方法。冷冻保存用于长期储存优良种质越来越多地被用于无性系农艺作物的长期保存，但对于像桉树这样的林木，尚未建立多样化遗传资源收集品的冷冻保存方法。我们报告了13个桉树品种离体茎尖培养种质收集品的成功冷冻保存，这些品种包括2个巨桉×赤桉品系、7个尾叶桉×巨桉品系、1个巨桉品系、2个巨桉×尾叶桉品系和1个赤桉品系。在两种冷冻保存方法的比较中，蔗糖敏感性限制了包埋脱水法的应用。然而，采用玻璃化法，所有13个品系在液氮冷冻保存后都有良好的存活率。根据品系不同，在植物玻璃化溶液2（PVS2）中处理30分钟后，液氮处理后的存活率在38%至85%之间。目前，所有13个品系的100个茎尖作为种质收集品进行长期保存。
