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1.
影响枣组培苗玻璃化的几个因素及其防治(简报)   总被引:5,自引:0,他引:5  
高温、高浓度细胞分裂素6-BA和低透气性均促进枣组培苗玻璃化现象的发生,改变以上3种因子可在一定程度上防止或减轻玻璃化的发生。  相似文献   

2.
几种因素对牙鲆胚胎玻璃化冷冻保存的影响   总被引:4,自引:0,他引:4  
鱼类胚胎冷冻保存技术还远没有成熟, 为了寻找最佳的鱼类胚胎玻璃化冷冻保存条件, 我们以牙鲆(Paralichthys olivaceus) 胚胎为例, 研究了影响鱼类胚胎玻璃化冷冻保存的几个主要因子: 玻璃化液、麦管直径、胚胎阶段、平衡时间及平衡温度、洗脱浓度和洗脱时间。发现: (1) 含有多种抗冻剂的玻璃化液PMDD(2% PVP), 玻璃化稳定, 脱玻璃化率较低, 适宜进行玻璃化冷冻; (2) 尾芽期胚胎较其他时期耐受力强, 平衡40 min就足以使玻璃化液渗透完全, 时间延长, 成活率显著降低, 各个时期的胚胎对温度都比较敏感, 0°C与4°C下平衡的成活率显著高于15°C; (3) 洗脱浓度和洗脱时间对胚胎成活率影响不大; (4) 根据优化的条件, 对牙鲆两个时期的胚胎进行超低温冷冻保存实验, 共成活4次, 获得成活胚胎8粒, 其中7粒孵化出健康的鱼苗。本文为鱼类胚胎冷冻保存技术的建立提供基础资料, 并显示了牙鲆胚胎玻璃化冷冻保存是可行的。  相似文献   

3.
保存活体的肺癌组织将为肺癌发病基因筛查和靶向药物筛选等体外实验研究提供更完整的样本信息. 本文对活体肺癌组织的玻璃化保存方法进行研究,首先采用针浸法玻璃化保存单块肺癌组织,对所需低温保护剂的浓度和平衡时间进行了优化;其次采用冻存管对多块肺癌组织样本进行玻璃化保存,对低温保护剂溶液体积以及平衡时间进行了优化;最后对慢速冷冻、不加低温保护剂快速冷冻、玻璃化冷冻3种冷冻方法的冻存效果进行比较并通过低温显微分析其冰晶损伤机理.结果表明,20% EG+20% DMSO+0.5 mol/L海藻糖作为低温保护剂,在平衡溶液和玻璃化溶液分别加载3 min和1 min时,针浸法和0.25 ml冻存管内玻璃化冻存,复苏后组织活力最高,分别约为79.96%与80.44%. 免疫组化显示玻璃化保存肺癌组织经过复苏后,相比慢速冷冻和无保护剂快速冷冻,组织结构损伤较小,组织内细胞TUNEL阳性表达较少. 低温显微结果表明,玻璃化保存组织内部及周围只出现少量细小冰晶,而慢速冷冻、快速冷冻组织皆出现明显冰晶.  相似文献   

4.
不同培养条件对光叶楮试管苗玻璃化的影响   总被引:2,自引:0,他引:2  
以MS为基本培养基,研究生长调节剂、蔗糖、琼脂、光照强度和温度等因素对光叶楮试管苗玻璃化的影响。结果表明,最佳的生长调节剂配比是1.0mg/L 6-BA、0.3mg/L NAA,蔗糖浓度3.5%,琼脂浓度0.35%,光照强度2 000 lx,培养温度25℃。该优化培养条件能有效防止玻璃化苗的发生,获得较高的增殖系数和较健壮的苗木,且能降低生产成本。  相似文献   

5.
梨芽离体快繁过程中玻璃化苗的发生与氧自由基胁迫的关系   总被引:12,自引:1,他引:11  
优质梨“苹博1号”的芽离体快繁过程中发生的玻璃化苗与其它植物的玻璃化苗相似,试验对其氧自由基清除酶活性和膜脂过氧化水平进行了研究。结果表明;正常试管苗和玻璃轩的过氧化物酶,过氧化氢酶,超氧化物歧化酶的活性和丙二醛含量普遍随培养时间而呈升高趋势;但玻璃化苗的后两种酶活性低于正常苗,可溶性蛋白含量也显著低于正常值,丙二醛水平却显著高于正常苗,提示梨快繁过程中有氧自由基的胁迫,而且玻璃化苗发生了更严重的  相似文献   

6.
鲈鱼胚胎的玻璃化冷冻保存   总被引:6,自引:0,他引:6  
本文对鲈鱼(Lateolabrax japonicus)胚胎进行了玻璃化冷冻保存研究,筛选出了浓度较低、玻璃化程度较稳定的5种玻璃化液,冷冻时形成玻璃化的概率在48.1%~100%,在35~43℃的水浴中解冻时保持玻璃化的概率在44.4%~63.0%;玻璃化液VSD2在解冻时保持玻璃化的概率最高。对鲈鱼神经胚、20对肌节胚、尾芽胚、心跳胚、出膜前胚在玻璃化液VSD2中的适应能力及适合于玻璃化冷冻的胚胎时期进行了比较,结果显示:不同时期胚胎对玻璃化液的耐受能力不同,鲈鱼神经胚耐受能力最低,心跳胚耐受能力最强,出膜前期胚次之,心跳胚和出膜前胚适合于进行玻璃化冷冻。对0.5mol/L蔗糖的洗脱时间进行了选择,结果显示,洗脱10~20min效果较好。利用玻璃化程度较好的VSD2对鲈鱼不同时期胚胎进行超低温(-196℃)冷冻,获得了2.1%~27.9%的透明胚。将鲈鱼心跳胚冷冻解冻后获2粒复活胚,培养至出膜期,成活42~50h;出膜前期胚在冷冻解冻后有1粒胚复活,并且孵化出鱼苗[动物学报49(6):843~850,2003]。  相似文献   

7.
油菜试管苗玻璃化发生机理的研究初探   总被引:1,自引:1,他引:0  
研究了影响甘蓝型油菜离体下胚轴再生过程中试管苗玻璃化产生的几个因子,试验发现:调整激素6-BA与NAA的浓度、增加蔗糖的浓度提高渗透势对玻璃化无影响;降低培养基中NH4 浓度不仅没有有效地防止玻璃苗的发生反而降低了芽的诱导率;采用牛皮纸代替聚乙烯塑料膜作为封口材料的措施受到了地区的限制,没有得到有效的统计数据;选择1%的琼脂浓度可使玻璃化率显著降低55%,但诱芽率在一定程度上降低了6.9%。  相似文献   

8.
玻璃化是指植物生长过程中出现的类似水浸状的半透明化生理病变现象, 常见于植物组织培养。玻璃化现象是限制组培技术发展和组培苗商业化生产的三大主要影响因素之一, 但是玻璃化的发生机制至今尚不清楚。以组培材料为对象研究玻璃化机制受多种人为因素的干扰, 而以非组织培养材料为研究对象, 避免了人为因素的干扰, 有助于从本质上揭示玻璃化发生的分子机制。该文综述了非组培苗因软木脂含量改变、表皮蜡质合成减少、细胞膜过氧化受损、离子或水分子跨膜运输受阻等导致植物玻璃化的机制, 以期为最终揭示植物玻璃化机制提供参考。  相似文献   

9.
研究了蓝莓试管苗玻璃化的显微结构、超微结构以及生理生化特性的影响。与正常试管苗相比,蓝莓玻璃化苗的茎、叶显微结构发生了明显的改变:叶片表皮细胞松散、不连续;气孔结构难以辨认;叶片增厚;缺少栅栏组织,海绵组织细胞间隙变大,部分细胞解体;茎的维管组织发育不良;亚显微结构观察发现,玻璃化苗叶肉细胞体积增大,细胞壁变薄;部分细胞缺少细胞核及线粒体;叶绿体数目减少,类囊体解体,缺乏淀粉体。玻璃化试管苗的生理生化特性也发生了显著的改变:玻璃化苗组织含水量显著增加;叶绿素、可溶性糖及可溶性蛋白含量显著降低;O2- 产生速率、H2O2积累量、MDA含量及相对电导率显著升高;活性氧清除酶系中POD活性显著升高,SOD和CAT活性显著降低;PAL活性下降。蓝莓玻璃化苗的形态结构异常,水分及物质代谢紊乱,活性氧清除能力降低,表明玻璃化与氧化胁迫相关。  相似文献   

10.
拟南芥悬浮细胞系的玻璃化法超低温保存   总被引:6,自引:1,他引:5  
悬浮培养细胞系是植物生理生化研究的好材料之一。为了保持细胞系的遗传稳定性,需要采用超低温保存技术。玻璃化法是一种不用程序降温仪的超低温保存技术。本文报道了从模式植物拟南芥建立悬浮细胞系并对其进行玻璃化法超低温研究。细胞经过合理的预培养处理和保护剂处理,直接投入液氮贮存。复温后的细胞能恢复生长,恢复生长的细胞保持着植株再生能力。国外,拟南芥悬浮细胞系的程序降温法保存和包埋脱水法保存已经报道,玻璃化法保存尚未见报道。  相似文献   

11.
满天星组织培养中克服玻璃化现象的初探   总被引:23,自引:0,他引:23  
张燕玲  姚军  王润珍  唐高凤   《广西植物》1997,17(3):246-248
通过在培养基中加入不同浓度的糖分、琼脂、细胞分裂素(BA和KT)、聚乙烯醇(PVA)进行试验研究,发现满天星玻璃苗的数量随着糖分及琼脂浓度增加而减少,但细胞分裂素对玻璃化现象无影响,而聚乙烯醇对满天星玻璃化现象有明显的抑制作用,其最适浓度为20g/L。  相似文献   

12.
Summary Variations of composition and consistency of the culture medium and time of exposure to growth regulators were assayed to optimize normal caulogenic response ofDigitalis obscura hypocotyls cultured in vitro. The effects of the culture conditions on physiologic changes related to vitrification of the regenerated plants were also investigated. Liquid medium increased the bud-forming capacity of the explants but induced buds failed to develop into shoots and showed symptoms of vitrification. On agar-solidified media, maximum multiplication rates were achieved with 0.7% agar. Increasing agar concentration reduced vitrification but lowered the propagation rate. Changes in the strength of the macronutrients of Murashige and Skoog did not significantly affect the bud-forming capacity of the explants. In contrast, a drastic inhibitory effect on both bud formation and shoot elongation was produced when NH4NO3 was omitted. Reduction of NH4NO3 to one-half or one-fourth of the level of the original formulation not only increased the bud-forming capacity ofD. obscura hypocotyls but also resulted in less vitrification. Modifications of time and method of exposure to growth regulators neither improved the multiplication rates nor overcame vitrification. Cardenolide content was lower in vitrified than in normal cultures and coincided with an overall reduction of photosynthetic pigments, lignin, and dry matter.  相似文献   

13.
Investigation was made on a use of microcrystal cellulose as a new and inexpensive gelling agent instead of agar. Microcrystal cellulose in concentration 20 % forms a suitable structure of nutrient medium for in vitro cultivation. The higher humidity in the culture container with microcrystal cellulose causes partial vitrification of Nicotiana tabacum L. plants, cv. Zlatna arda. It is proved by reduced chlorophyll content, changes in protein synthesis and strongly reduced isoenzyme spectrum of peroxidase.  相似文献   

14.
Summary Normal nodal segments ofPetunia hybrida were grown on Murashige and Skoog salts containing varied levels of Gelrite and sucrose. Higher concentrations of Gelrite decreased vitrification while increased sucrose concentrations promoted vitrification. Leaves of vitreous plants had higher levels of reducing sugars and sucrose but lower or undetectable levels of inositol as compared to normal plants. Normal plants on medium void of inositol have the ability to synthesize inositol and maintain levels equal to that found in plants from inositol containing media.  相似文献   

15.
植物种质的玻璃化超低温保存   总被引:19,自引:0,他引:19  
植物种质的玻璃化超低温保存技术已受到广泛重视。玻璃化法主要由装载、玻璃化保护液脱水、降温、复温、洗涤这5个环节构成。目前已对百余种植物进行过玻璃化冻存研究,但主要应用于高等植物,而用该法保存藻类获得成功的报道很少。将玻璃化法用于某些藻类种质的冻存将会有广阔的应用前景。  相似文献   

16.
Summary Aeration of tissue cultured rare Australian plantsConostylis wonganensis S.D. Hopper (Haemodoraceae);Diplolaena andrewsii Ostenf.;Drummondita ericoides Harvey (Rutaceae);Eremophila resinosa F. Muell. (Myoporaceae);Eucalyptus ‘graniticola’ (Myrtaceae);Lechenaultia pulvinaris C. Gardner (goodeniaceae); andSowerbaea multicaulis E. Pritzel (Liliaceae) has been found to reduce vitrification in sensitive species as well as significantly improving shoot quality and transfer to soil in most study species. A simple 7-mm hole with a double-layer insert of filter paper in the polypropylene screw lids of the culture vessel decreased shoot vitrification over a 4-wk culture period. The method has implications for facilitating the tissue culture of other rare Australian plants and reducing the occurrence of this developmental abnormality.  相似文献   

17.
李海刚  孔祥生 《生物学杂志》2010,27(5):35-37,42
以MS为基本培养基,研究品种、琼脂、6-BA、大量元素、蔗糖、光照等因素对牡丹试管苗玻璃化的影响。结果显示:不同品种的牡丹试管苗在组织培养的过程中,出现不同程度的玻璃化,增加培养基中琼脂浓度、降低细胞分裂素6-BA的浓度、降低大量元素的浓度和增加光照强度或采用自然光照均可以有效地降低试管苗玻璃化。  相似文献   

18.
Vitrification is considered a viable method for cryopreservation of ovarian tissue and selection of methods that minimize follicular damage is important. The objective of the present study was to evaluate the effects of two vitrification methods on ovarian tissue morphology, preantral follicles survival rate during in vitro culture, and relative expression of genes associated with oocyte maturation and cumulus expansion. Ovaries from 12-day-old mice were vitrified in media containing ethylene glycol, dimethyl sulphoxide, and sucrose. Before plunging in liquid nitrogen, ovaries were first loaded into an acupuncture needle (needle immersion vitrification [NIV]) or placed on a cold steel surface for 10 to 20 seconds (solid surface vitrification [SSV]). The integrity of the ovarian tissue was well-preserved after vitrification and was similar controls. Follicle viability in the SSV group was lower (P < 0.05) than in the control group after 6 days of culture and the NIV group after 10 day of culture. Follicle viability after 12 day of culture was 92.8%, 82.1%, and 58.4% in control, NIV, and SSV groups, respectively. Bmp15, Gdf9, BmprII, Alk6, Alk5, Has2, and Ptgs2 gene expression patterns were similar among groups. However, the level of gene expression in the vitrification groups during Days 6 to 10 were higher compared with the control group. In conclusion, ovarian tissue morphologic integrity was well-preserved, regardless of the vitrification method. Vitrification using the needle immersion method resulted in greater follicular survival after 12 day of culture than the SSV method. Gene expression patterns during culture did not seem to explain the reduced survival rate observed in the solid surface group.  相似文献   

19.
The objective of this study was to compare the overall efficiency, measured by in vitro embryonic survival, and practical value of bovine in vitro embryo production, biopsy, vitrification, and direct transfer technology using 2 different manipulation methods for biopsy. Slaughterhouse-derived oocytes were matured in vitro, fertilized (Day 0) with frozen-thawed, Percoll-separated spermatozoa and cultured on a granulosa cell monolayer. In Experiment 1, one or two blastomeres were expelled from Day 4 embryos by mechanical force through a hole made by partial zona dissection. Using a darning needle hole system for individual culture of biopsied embryos from Day 4 to Day 7.5, the blastocyst per oocyte rate was 50%, and 76% of the blastocysts survived subsequent vitrification and direct in-straw rehydration. Attempts to increase the cell number of the biopsies by further in vitro culture were unsuccessful. In Experiment 2, Day 7 and Day 8 embryos were manually biopsied before or after vitrification. When biopsy was performed before vitrification, 98% of the embryos survived manipulation, and 86% of these re-expanded after vitrification and in-straw dilution. Biopsy after vitrification was less efficient, since only 69% of the embryos survived both processes. The cumulative efficiency of embryo production, Day 7.5 biopsy and vitrification--in-straw direct rehydration was lower (P < 0.001) than that of Day 4 biopsy and Day 7.5 vitrification (29 vs 38%, respectively). However, a Day 7.5 biopsy may have the more practical application since the size of the biopsy is larger and the process is not as time-consuming as the long-term individual culture of the biopsied embryos.  相似文献   

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