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

我们设计了一种在非无菌室中更换体外培养基的方法，保证理论上能够保持膜培养的准确性，反渗透（R.O.）膜是一种允许水自由通过的离子膜。除大分子外，磷是地球上所有生命所必需的元素，在蔗糖溶液中，通过消除磷来防止微生物的繁殖。通过使用该膜和不腐烂的蔗糖溶液，我们开发了用于更换体外培养基的系统。在该系统中，容器漂浮在不腐烂的蔗糖溶液上，用于氢氧化铝，底部被密封。植物和磷酸盐（熔融磷酸镁）被放置在该容器中的 R.O. 膜上。通过额外的灭菌氢氧化铝涂层，几乎可以防止磷酸盐泄漏，从而防止蔗糖溶液腐烂，并保持植物的营养和蔗糖供应超过两个月。我们还设计了康乃馨、马铃薯、草莓的芽尖在非无菌室中进行芽尖培养和继代培养的简化方法。将百合和结冷胶接种到高压灭菌后的试管中，然后在每个试管中加入少量稀释的次氯酸钠溶液。加入次氯酸钠溶液400μl（含活性氯5～50mg/l），覆盖80%以上的芽尖。所有受检物种均在无微生物污染的培养基上存活并生长良好。这两种方法的结合对于体外培养的简化可能非常有用。