短期培养对玻璃化冷冻小鼠2-细胞胚胎低渗抵抗力的修复作用

为研究短期培养对玻璃化冷冻小鼠2-细胞胚胎低渗抵抗力的修复作用,对玻璃化冷冻小鼠2-细胞胚胎解冻后进行短期(2～4 h)培养,然后使用20%的低渗PBS液于25℃条件下处理20 min,观察48 h发育率、囊胚率和移植后妊娠率、产仔率.获得93%的48 h发育率和48%的囊胚率,显著高于低渗处理前未培养组(47%和9%)(P＜0.05).证明解冻后的短期培养中可以有效修复玻璃化冷冻对小鼠2-细胞胚胎低渗抵抗力的损伤.     

香蕉茎尖的玻璃化法超低温保存及其植株再生

 以香蕉(Musa spp. ) 为试材, 对其离体培养茎尖玻璃化法超低温保存影响因素进行研究。结果表明, 不定芽在MS + 3.0～5.0 mg/L 6-BA + 0.1 mg/L NAA的培养基上分化较好。香蕉茎尖超低温保存较佳体系是: 2.0～3.0 cm的茎尖在含0.4 mol/L蔗糖培养基上预培养2 d, 剥取带1～2个叶原基的茎尖(长1.0～1.5 mm) , 室温(25℃) 下装载液(MS + 2 mol/L甘油+ 0.4 mol/L蔗糖) 装载20～30 min, 然后用玻璃化溶液( PVS2 ) 于0℃下处理40 min, 换1次PVS2后迅速投入液氮。保存至少1 h后, 在40℃水浴中化冻90 s, 用1.2 mol/L蔗糖培养液洗涤2次, 每次10 min, 然后转入含0.3 mol /L蔗糖的MS培养基上,暗培养10～15 h后转移到含0.5 mg/L 62BA的MS培养基中, 暗培养1周后转移到正常光下, 3个香蕉品种(巴西蕉、广东香蕉2 号、广东粉蕉1 号) 的成活率分别为75.9%、40.0%和69.6% , 再生率分别为63.4%、35.0%和63.4%。再生植株生长和分化正常, 生根后可移栽成活。     

Cryopreservation of shoot apices of hawthorn in vitro cultures originating from East Asia

The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species.     

植物组织培养中玻璃化现象的研究进展

从植物组织培养中玻璃化的概念及研究意义出发，在比较玻璃化苗在形态解剖上和生理生化的异常变化的基础上分析了试管苗玻璃化发生的因素和必要的预防措施，综述近年来国内外在以上问题上的研究进展，并对此问题提出一些看法。     

Ultrastructural Morphology and Nuclear Maturation Rates of Immature Equine Oocytes Vitrified with Different Solutions and Exposure Times

Ultrastructural morphological injuries and maturation rates were investigated in equine oocytes exposed to vitrification solutions (VS) containing synthetic ice blockers (SIBs) during different exposure times. In experiment 1, compact cumulus-oocyte complexes (COCs; n = 30) were randomly allocated to treatments: (1) fresh fixed (control); (2) VS-1 (1.4 M dimethyl sulfoxide [DMSO] + 1.8 M ethylene glycol [EG] + 1% SIB) for 3 minutes of equilibrium time and VS-2 (2.8 M DMSO + 3.6 M EG + 0.6 M sucrose + 1% SIB) for 1 minute (Eq-long); and (3) VS-1 for 1.5 minutes and VS-2 for 30 seconds (Eq-short). In experiment 2, compact (n = 248) and expanded (n = 264) COCs were evenly distributed to the following treatments: (1) immediate maturation in vitro (control); (2) vitrification using the Eq-short protocol as in experiment 1; and (3) vitrification using a stock solution containing 2.8 M formamide, 2.8 M DMSO, 2.7 M EG, 7% polyvinylpyrrolidone, and 1% SIB (Eq-short-mod). More (P < .02) oocytes with normal ultrastructural morphology were seen in fresh control and Eq-short groups than in Eq-long group. Metaphase-II (MII) rates were higher (P < .05) for oocytes with expanded cumulus than compact cumulus in the control group, and higher (P < .05) for oocytes with expanded cumulus than compact cumulus in Eq-short and Eq-short-mod groups. No difference in MII rates was detected among groups within each type of COC. In conclusion, reduction of exposure time to VS better preserved oocyte ultrastructural features, and MII rates were higher for vitrified oocytes with expanded cumulus. This study advances our knowledge on potential alternatives for vitrification of immature equine oocytes.     

草莓组织培养的研究概述

组织培养技术是解决植株快速繁殖的常用技术之一,目前我国草莓组织培养技术主要有茎段、叶片、叶柄、花药组织培养等方法,本文重点对草莓的组织培养方法进行了叙述,并对褐化、污染和玻璃化现象的原因及解决方法进行了分析。     

马哈利樱桃离体茎尖超低温保存的研究

用玻璃化法超低温保存马哈利樱桃；切取经５℃低温驯化３０d的离体茎尖置于固体培养基 （ＭＳ＋０．７mol/Ｌ蔗糖）预培养２４h，在含１mＬ５０％甘油＋５０％蔗糖的保护液中处理８０min后浸入液氮。２５℃水浴快速化冻２min，然后接种到标准培养基上。２１d后调查，茎尖存活率达８４．６％。     

苹果离体培养中试管苗玻璃化现象发生机理的探讨

试管苗玻璃化现象是植物组织培养中普遍发生的一种生理病变。其形态结构变异失常,生化成分及酶学特性也出现明显变化。导致这种现象的原因,初步认为是培养环境中的某种胁迫条件诱导试管产生应激乙烯,乙烯的直接或间接作用引起玻璃化现象的发生。     

牛胚胎玻璃化冷冻技术的发展和应用概况

胚胎冷冻保存是动物胚胎工程的重要组成部分，它是胚胎移植技术在生产中推广应用的重要基础。和现在通用的常规冷冻法相比，玻璃化冷冻具有用时短，所需设备少和操作简单等优点，成为目前国内外胚胎冷冻保存领域发展新方向。本文就牛胚胎玻璃化冷冻的概念、方法和国内外发展状况做一简要综述。