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Continous change of culture medium in a vessel unsterilized condition using reverse osmosis membrane

使用反渗透膜在未灭菌的容器中连续更换培养基

基本信息

批准号：
10660024
负责人：
MIZUTA Youichi
金额：
$ 2.56万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660024/
关键词：
functional membrane reverse osmosis membrane sucrose in vitro Phosphate Alminium chlorine 逆漫透謹

项目摘要

We designed a method for the changing in vitro culture medium in a non-sterile room. It is sure to be able to keep theoretically accurate with membrane culture which Reverse osmosis (R.O.) membrane is a semipermeable membrane which allows water to pass freely and ionic compounds slowly except large molecules. Phosphorus is essential element for all lives on earth. In sucrose solution, multiplication of microbs are prevented by elimination of phosphorus. The phosphorus was eliminated by refractoriness of aluminum phosphate, by mixing of aluminum hydroxide.By using this membrane and no-rotting sucrose solution, we developed system for changing in vitro culture medium. In this system, vessel floats on the no-rotting sucrose solution for aluminum hydroxide. The bottom is sealed with the R.O. membrane. Plant and phosphate-sourse (Fused magnesium phosphate) are sterilized and placed on the R.O. membrane in this vessel. Water and nutrients are supplied to the plant from the solution through t … More he R.O. membrane. By additional sterilized-aluminum hydoroxide coat, phosphate leakage ptetty nearly were prevented, so sucrose solution were prevented from rotting and plants were kept to nutrition and sucrose supplying for more than two month. But, plant growth were less than normal in vitro plants.We designed a simplified method for the shoot apex culture and subculture in a non-sterile room, too. Shoot apexes of carnation, potato, strawberry and lily were inoculated onto the medium containing Gellan Gum and nutrient salts in test tubes after autoclaving, in a non-sterile room. Then a small amount of diluted sodium hypochlorite solution was added into each test tube. In the test tubes to which 200 - 400 μl of sodium hypochlorite solution (containing 5 - 50mg/l active chlorine) was added, more than 80% of the shoot apexes of all species examined survived and grew well on the medium free of microbial contamination.Combination of this two methods may be very useful at in vitro culture simplification. Less

我们设计了一种在无菌房间中更换体外培养基的方法。用反渗透(R.O.)的膜培养方法在理论上保证了其准确性。膜是一种半透膜，除了大分子外，水可以自由通过，离子化合物可以缓慢通过。磷是地球上所有生命所必需的元素。在蔗糖溶液中，磷的去除阻止了微生物的繁殖。利用磷酸铝的耐火性和氢氧化铝的混合来除磷，并利用该膜和不腐烂的蔗糖溶液，开发了体外培养基更换系统。在这个系统中，容器漂浮在不腐烂的氢氧化铝蔗糖溶液上。底部用R.O.薄膜密封。植物和磷源(熔融磷酸镁)经过灭菌，放置在该容器的R.O.膜上。水和养分通过t…从溶液中提供给植物。更多的是他的R.O.膜。通过添加无菌氢氧化铝涂层，几乎防止了磷酸盐渗漏，防止了蔗糖溶液腐烂，使植物保持营养和蔗糖供应两个月以上。但是，试管苗的生长却低于正常植株。我们还设计了一种简化的方法，用于茎尖培养和无菌室内继代培养。香石竹、马铃薯、草莓和百合的茎尖经高压灭菌后，接种到含有结冷胶和营养盐的试管中，在无菌室内进行。然后在每个试管中加入少量稀释的次氯酸钠溶液。在含有2 0 0~4 0 0μL的次氯酸钠溶液(含5~50 mg/L有效氯)的试管中，供试植物80%以上的茎尖在无微生物污染的培养基上存活和生长良好。这两种方法的结合在试管外植体的简化中是非常有用的。较少