Development of PVS-Based Vitrification and Encapsulation–Vitrification Protocols
Development of PVS-Based Vitrification and Encapsulation–Vitrification Protocols
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DOI：
10.1007/978-0-387-72276-4_3
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发表时间：
2008
期刊：
影响因子：
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通讯作者：
A. Sakai;D. Hirai;T. Niino
中科院分区：
文献类型：
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作者：
A. Sakai;D. Hirai;T. Niino
超低温保存是为后代长期储存植物遗传资源的一个非常重要的工具，只需要最小的空间和维护。随着人们对植物基因工程的兴趣日益增加，具有独特属性的培养细胞和体细胞胚的保存变得更加重要。最近，据报道，低温保存提供了真实的希望，以提高濒危和稀有植物的保护（Toullit 1995; Toullit和狄克逊1996）。本章介绍了使用植物玻璃化溶液2（PVS2）成功玻璃化的协议，并强调了一些因素有助于高水平的LN后恢复。发展一种简单可靠的低温保存方法将使培养细胞、分生组织和体细胞胚的广泛储存成为可能。20世纪90年代开发了玻璃化法（包括玻璃化溶液（Langis et al. 1990; Sakai et al. 1990; Yamada et al. 1991））和冷冻-脱水技术（法布尔和Dereuddre 1990），此后冷冻保存物种的数量显著增加（Sakai 1995，1997; Engelmann和Takagi 2000）。Steponkus及其同事提出了一种使用乙二醇基玻璃化溶液和法国吸管的玻璃化程序。他们报道了石竹和菊花（Langis et al. 1990; Schnabel-Preikstas et al. 1992）和马铃薯（Lu and Steponkus 1994）的成功低温保存。本章将概述由Sakai及其同事开发的PVS2的开发和使用。
Cryopreservation is a very important tool for the long-term storage of plant genetic resources for future generations, requiring only a minimum of space and maintenance. With increasing interest in the genetic engineering of plants, the preservation of cultured cells and somatic embryos with unique attributes is assuming greater importance. Recently, cryopreservation was reported to offer real hope for enhancing the preservation of endangered and rare plants (Touchell 1995; Touchell and Dixon 1996). This chapter describes protocols for successful vitrification using plant vitrification solution 2 (PVS2), and highlights some of the factors contributing to high levels of post-LN recovery. The development of a simple and reliable method for cryopreservation would allow the widespread storage of cultured cells, meristems, and somatic embryos. Vitrification involving vitrification solutions (Langis et al. 1990; Sakai et al. 1990; Yamada et al. 1991) and encapsulation-dehydration techniques (Fabre and Dereuddre 1990) were developed in the 1990s, and the number of cryopreserved species has increased markedly since then (Sakai 1995, 1997; Engelmann and Takagi 2000). A vitrification procedure using an ethylene glycol-based vitrification solution and French straws was presented by Steponkus and colleagues. They reported successful cryopreservation ofDianthusandChrysanthemum(Langis et al. 1990; Schnabel-Preikstas et al. 1992), and potato (Lu and Steponkus 1994). This chapter will outline the development and uses of PVS2 developed by Sakai and associates.