[SIZE=+1]Steps in Tissue Culture[/SIZE]
(images courtesy of Dr. Dan Lineberger, aggie-horticulture.tamu.edu/tisscult/microprop/microprop.html)
Explant: Cut-out Plant Tissue and Place in Tissue Culture Container
The first step is to obtain what is called and explant. This means to simply cut-out a very small piece of leaf or stem tissue, or even isolate individual cells, and place them in a tissue culture container. The tissue has to be sterilized so it will not have any contaminating bacteria or fungus. It is then placed inside the tissue culture contain on a gel called agar. In the agar is dissolved all the sugar, nutrients and hormones the plant needs.
Explants can be pieces of any part of the plant (leaves, stems, flowers, etc.),
or even individual isolated cells.
Multiplication: Tissue Grows and Produces Small Plants
The tissue will begin to grow. It may make a big blob of tissue called callus, or it may make new shoots directly from the explant tissue that was inserted in the container.
A mass of callus tissue is formed that is just starting to make new plantlets.
New plantlets (shoots with leaves) are forming.
If the conditions are right a small "forest" of plants
will develop in the tissue culture container.
Rapid Multiplication by Transfer of Cultures
Once the plantlets start developing, some can be removed and placed in new tissue culture containers. Thus, another "forest"' of plants is produced. This results in a rapid multiplication of the cultures and many thousand of plants can be produced in a few months.
Some of the small plantlets can be removed and transferred to new tissue culture
containers. These will produce more shoots and fill the container.
Transplanting
When the plantlets are large enough, they can be removed from the tissue culture container and transferred into pots with potting soil. The young plants are growth in a greenhouse just like you would any young seedling or cutting.
When the small plant clones are removed from the culture containers, they must be transplanted into some type of acclimation container or kept under a mist system until the acclimate to the ambient environment.
After acclimation, the young plants can be transplanted
and grown in pots in a greenhouse to produce new plants.