Helmut79
Well-Known Member
I would like to have your opinion about this technique.
Does it work well? Is it worth the hassle?
http://cals.arizona.edu/pubs/garden/mg/propagation/grafting.html#Top
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Plant Tissue Culture for the Home
Although technical procedures for aseptic culture of plant cells, tissues, and organs are as diverse as the plant material on which they are practiced, a simplified general procedure can be followed in the home. All that is needed are a few basic supplies which can easily be obtained. The procedures outlined in this section can be used in the home to propagate various species of plants, both easy (African violets, coleus, chrysanthemums) and difficult (orchids, ferns, weeping figs) to propagate.
Medium Preparation
For 2 pints of tissue culture medium, mix the following ingredients in a 1-quart home canning jar:
Fill the jar with distilled or deionized water. If purified water is not available, water that has been boiled for several minutes can be substituted. Shake the mixture and make sure all materials have dissolved.
Baby food jars with lids, or other heat-resistant glass receptacles with lids can be used as individual culture jars. They should be half filled with cotton or paper to support the plant material. The medium should be poured into each culture bottle to the point where the support material is just above the solution.
When all bottles contain the medium and have the lids loosely screwed on, they are ready to be sterilized. This can be done by placing them in a pressure cooker and sterilizing them under pressure for 30 minutes or placing them in an oven at 320oF for 4 hours. After removing them from the sterilizer, place them in a clean area and allow the medium to cool. If the bottles will not be used for several days, wrap groups of culture bottles in foil before sterilizing and then sterilize the whole package. Then the bottles can be removed and cooled without removing the foil cover. Sterilized water, tweezers, and razor blades, which will be needed later, can be prepared in the same manner.
Plant Disinfection and Culture
Once the growing medium is sterilized and cooled, plant material can be prepared for culture. Because plants usually harbor bacterial and fungal spores, they must be cleaned (disinfected) before placement on the sterile medium. Otherwise, bacteria and fungi may grow faster than the plants and dominate the culture.
Various plant parts can be cultured, but small, actively growing portions usually result in the most vigorous plantlets. For example, ferns are most readily propagated by using only 1/2 inch of the tip of a rhizome. For other species, 1/2 to 1 inch of the shoot tip is sufficient. Remove leaves attached to the tip and discard. Place the plant part into a solution of 1 part commercial bleach to 9 parts water for 8 to 10 minutes. Submerge all plant tissue in the bleach solution. After this time period, rinse off excess bleach by dropping the plant part into sterile water. Remember, once the plant material has been in the bleach, it has been disinfected and should only be touched with sterile tweezers.
After the plant material has been rinsed, remove any bleach-damaged tissue with a sterile razor blade. Then remove the cap of a culture bottle containing sterile medium, place the plant part onto the support material in the bottle making sure that it is not completely submerged in the medium, and recap quickly.
Transferring should be done as quickly as possible in a clean environment. Therefore, scrub hands and counter tops with soap and water just before beginning to disinfest plant material. Rubbing alcohol or a dilute bleach solution can be used to wipe down the working surface.
After all plants have been cultured, place them in a warm, well-lit (no direct sunlight) environment to encourage growth. If contamination of the medium has occurred, it should be obvious within 3 to 4 days. Remove and wash contaminated culture bottles as quickly as possible to prevent the spread to uncontaminated cultures.
When plantlets have grown to sufficient size, transplant them into soil. Handle as gently as possible because the plants are leaving a warm, humid environment for a cool, dry one. After transplanting, water the plants thoroughly and place them in a clear plastic bag for several days. Gradually remove the bag to acclimate the plants to their new environment; start with one hour per day and gradually increase time out of the bag over a two-week period until the plants are strong enough to dispense with the bag altogether.
Does it work well? Is it worth the hassle?
http://cals.arizona.edu/pubs/garden/mg/propagation/grafting.html#Top
'''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
Plant Tissue Culture for the Home
Although technical procedures for aseptic culture of plant cells, tissues, and organs are as diverse as the plant material on which they are practiced, a simplified general procedure can be followed in the home. All that is needed are a few basic supplies which can easily be obtained. The procedures outlined in this section can be used in the home to propagate various species of plants, both easy (African violets, coleus, chrysanthemums) and difficult (orchids, ferns, weeping figs) to propagate.
Medium Preparation
For 2 pints of tissue culture medium, mix the following ingredients in a 1-quart home canning jar:
- 1/8 cup sugar
1 teaspoon all-purpose, soluble fertilizer mixture. Check the label to make sure it has all of the major and minor elements, especially ammonium nitrate. If the latter is lacking, add 1/3 tsp. of a 35-0-0 soluble fertilizer
- 1 tablet (100 mg) of inositol (myo-inositol) which can be obtained at most health food stores 1/4 of a pulverized vitamin tablet which has 1 to 2 mg of thiamine
- 4 Tablespoons coconut milk (cytokinin source) drained from a fresh coconut. The remainder can be frozen and used later.
- 3 to 4 grains (1/400 teaspoon) of a commercial rooting compound which has 0.1 active ingredient IBA
Fill the jar with distilled or deionized water. If purified water is not available, water that has been boiled for several minutes can be substituted. Shake the mixture and make sure all materials have dissolved.
Baby food jars with lids, or other heat-resistant glass receptacles with lids can be used as individual culture jars. They should be half filled with cotton or paper to support the plant material. The medium should be poured into each culture bottle to the point where the support material is just above the solution.
When all bottles contain the medium and have the lids loosely screwed on, they are ready to be sterilized. This can be done by placing them in a pressure cooker and sterilizing them under pressure for 30 minutes or placing them in an oven at 320oF for 4 hours. After removing them from the sterilizer, place them in a clean area and allow the medium to cool. If the bottles will not be used for several days, wrap groups of culture bottles in foil before sterilizing and then sterilize the whole package. Then the bottles can be removed and cooled without removing the foil cover. Sterilized water, tweezers, and razor blades, which will be needed later, can be prepared in the same manner.
Plant Disinfection and Culture
Once the growing medium is sterilized and cooled, plant material can be prepared for culture. Because plants usually harbor bacterial and fungal spores, they must be cleaned (disinfected) before placement on the sterile medium. Otherwise, bacteria and fungi may grow faster than the plants and dominate the culture.
Various plant parts can be cultured, but small, actively growing portions usually result in the most vigorous plantlets. For example, ferns are most readily propagated by using only 1/2 inch of the tip of a rhizome. For other species, 1/2 to 1 inch of the shoot tip is sufficient. Remove leaves attached to the tip and discard. Place the plant part into a solution of 1 part commercial bleach to 9 parts water for 8 to 10 minutes. Submerge all plant tissue in the bleach solution. After this time period, rinse off excess bleach by dropping the plant part into sterile water. Remember, once the plant material has been in the bleach, it has been disinfected and should only be touched with sterile tweezers.
After the plant material has been rinsed, remove any bleach-damaged tissue with a sterile razor blade. Then remove the cap of a culture bottle containing sterile medium, place the plant part onto the support material in the bottle making sure that it is not completely submerged in the medium, and recap quickly.
Transferring should be done as quickly as possible in a clean environment. Therefore, scrub hands and counter tops with soap and water just before beginning to disinfest plant material. Rubbing alcohol or a dilute bleach solution can be used to wipe down the working surface.
After all plants have been cultured, place them in a warm, well-lit (no direct sunlight) environment to encourage growth. If contamination of the medium has occurred, it should be obvious within 3 to 4 days. Remove and wash contaminated culture bottles as quickly as possible to prevent the spread to uncontaminated cultures.
When plantlets have grown to sufficient size, transplant them into soil. Handle as gently as possible because the plants are leaving a warm, humid environment for a cool, dry one. After transplanting, water the plants thoroughly and place them in a clear plastic bag for several days. Gradually remove the bag to acclimate the plants to their new environment; start with one hour per day and gradually increase time out of the bag over a two-week period until the plants are strong enough to dispense with the bag altogether.
