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

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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