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

我们设计了一种在非无菌室中改变体外培养基的方法。反渗透膜是一种半透膜，除大分子外，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，磷的消除阻止了微生物的繁殖。通过与氢氧化铝混合，利用磷酸铝的耐火性去除磷。利用该膜和不腐烂的蔗糖溶液，研制了体外培养基更换系统。在该系统中，容器浮在不腐烂的氢氧化铝蔗糖溶液上。底部用R.O.膜密封。植物和磷酸盐源（熔融磷酸镁）被消毒并放置在该容器的R.O.膜上。水和营养物质从溶液中通过R.O.膜提供给植物。通过加装灭菌氢氧化铝涂层，几乎可以防止磷酸盐的泄漏，防止蔗糖溶液的腐烂，使植株保持营养和蔗糖供应两个月以上。但是，离体植株的生长却低于正常植株。我们还设计了一种在非无菌室进行茎尖培养和继代培养的简化方法。将康乃馨、马铃薯、草莓和百合的茎尖接种到含有结冷胶和营养盐的培养基上，在非无菌室中进行高压灭菌。然后在每个试管中加入少量稀释的次氯酸钠溶液。在次氯酸钠溶液（含活性氯5 ~ 50mg/l）浓度为200 ~ 400 μl的试管中，所有被检物种的茎尖在无微生物污染的培养基上成活率均在80%以上，生长良好。这两种方法的结合可能对体外培养简化非常有用。少

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