FLASKING: Reflasking with Kappa Carrageenan (?) as a Substitute for Agar

Orchid Doctor Propagation

by Robert M. [Bert] Hamilton (Compiler)

Originally published in The Orchid Doctor in 1980 and 1988

Posted by Sys Admin about 4 years ago.

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Extract from red seaweeds is used in the food industry and cosmetics; experiments with it as an orchid medium for flasking culture and tested comparatively with standard agar media. A83-1174
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More The Orchid Doctor Articles under the Heading Flasking

Flasking Comments
FLASKING: Air Supply to Seedlings For cymbidium seedlings in flask, air exchange must always be provided. OA84-126 0
FLASKING: Banana Added to Prepared Agar Ripe or green banana is homogenized in a kitchen blender; add 5% of it to the medium. A74-792 0
FLASKING: Best Procedure to Avoid Reflasking Sow the seed thinly in the original flask. A75-35 0
FLASKING: Black Substance in the Culture Medium Phalaenopsis seedlings are notorious for exuding a black material from the roots; it is not a contaminant but in high concentrations impedes growth. F82-175 0
FLASKING BOX: Cheap and Adequate Made from a cardboard box, saran wrap and aluminium foil; diagrams; refer to A69-286 0
FLASKING BOX: Made of Plexiglas For details of compact, workable unit, illustration, refer to A86-820 0
FLASKING CASE: Laminar Flow Filter To remove fungus spores the filter should be four microns; for bacteria less than one micron. A74-502 0
FLASKING CASE: Made Out of a Corrugated Carton For directions and illustration, refer to OD68-77 0
FLASKING CASE: Made Out of a Foam Picnic Cooler How to make and use; refer to A77-929+ 0
FLASKING: Culture Medium for Terrestrial Cypripediums Excellent results reported with: Hyponex 3 grams; Difco-Bacto peptone or tryptone 2 grams, sucrose 35 grams, agar 15 grams, water 1000 ml. and pH adjusted to 5.0 A63-354+; A63-927 0
FLASKING: Delayed Contamination It is possible when a pathogen was introduced to the sterile flask and remained lodged near the opening; it would take severa1 weeks to reach the agar medium. F82-175 0
FLASKING: Detailed Working Instructions with Illustrations and Much Detail Refer to A60-597+; A64-759+; A70-599+; A75-109+; OD68-204+ 0
FLASKING: For the Amateur Australian style, refer to AU79-133+ 0
FLASKING: Improved Methods of Germination Sterile medium is achieved by decontaminating glassware with Physan 20 solution and adding .005% hydrogen peroxide into the medium just before pouring it into flasks; sterilize seed by soaking it 24 hours in sugar water, then in 3% solution hydrogen peroxide; other limits, refer to A85-178 0
FLASKING: Kitchen Style A solution to many problems was found in the use of a clear plastic bag 10 x 12 inches; two holes were cut to fit over the openings of the flasks at the sealed end and the work was done through the open end with success; for particulars refer to Ca86(3)-25; see also Ca82(6)-4 0
FLASKING: Methods and Materials Apparatus and growth materials required; preparation of seed; aids for simplifying the process; for green capsule and ripe seed; refer to OR81-149; Ca82(6)-4; OBII-243 0
FLASKING: Prepared Flasks for Later Use Keep them on hand for about one week only; but can be kept for about two months if stoppers are covered with tightly fitting aluminum foil. RMH 0
FLASKING: Proliferation of Tissue Reasons are not clear and no anti-proliferations have been mentioned but certain additives such as banana homogenaate may be useful; for references ref.to A85-1334 0
FLASKING: Reflasking Airtight capping is not advised as the seedlings can be inhibited by it. A77-314 0
FLASKING: Reflasking Alternative Use a 2-liter plastic pop bottle; heat the bottom stand in hot water to soften the adhesive and pull it off, cut off the top section of the pop bottle and invert it into the stand portion to form a "terrarium"; boil a pan of uomilled sphagnum moss as growing medium for the seedlings. A85-1331; OEE86Jan-15 0
FLASKING: Replate Medium Use the same germinating medium for replating; the Knudson-C formula can be used for both. A70-631 0
FLASKING: Roots Do Not Penetrate the Medium This may happen because the stopper is too tight and eliminates gas exchange between inside and outside of the flask. F82-175 0
FLASKING: Roots Grow Above Agar If exchange of air is not provided roots will reach upward instead of into the agar media. F72-136 0
FLASKING: Roots Grow Upon Agar Most roots grow on the surface; at the sides some grow to the bottom of the flask. A65-832 0
FLASKING: Rubber Stoppers Plugged with Cotton The cotton should be just firm; foil is not necessary except during the sterilization process and for a day or two after until the cotton dries. A75-192 0
FLASKING: Stopper Type Preferred Bakelite screw caps are good; for the long run required by orchids rubber with cotton in the center hole is preferred; Mason jars with screw caps are also satisfactory. A75-192 0
FLASKING: Transflasking Improvements They result from the replenishment of nutrients and of pH adjustments; probably the increase in water plays a part; more space influences all factors. A68-524 0

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