How to Breed a Plant In Vitro

In vitro plants develop specific characteristics that are controlled by the grower.

There are many ways to propagate plants but some do not produce the desired traits unless the embryos are handpicked for growth. In vitro reproduction enables researchers to create different breeds and increase genetic diversity. In vitro is a laboratory process that develops pre-breeding material. The process grows a tissue culture from the cells of a plant called an explant. The culture grows on a source of food called agar and becomes a plantlet. In vitro or micropropagation plant breeding also requires careful temperature and lighting control as well as sterile conditions. The harvested tissue cells from the mother plant must be healthy or the plantlet will not form normally.

Things You'll Need

• Agar
• Sugar
• Spoon
• Sauce pan
• Petri dishes with lids
• Bleach
• Distilled water
• Spray bottle
• Mother plant
• Razor blade
• Tweezers
• Sterile bowl
• Sterile gloves
• Rooting Hormone
• Paintbrush
• Pot
• Peat moss

Instructions

1. Pour agar into a saucepan. Mix in 1 to 2 tbsp. of sugar. Let the two items melt together on low heat, stirring often. The amount will depend on the size of culture you will be taking. Let the agar and sugar mix cool slightly and pour it into petri dishes.

2. Make a 10 percent solution of bleach and distilled water for sterilizing items as you continue. Sterilize a razor blade and harvest the explant. Excise a piece of healthy leaf tissue no larger in diameter than a pencil eraser. Dip it into the bleach-and-water solution repeatedly using tweezers to hold it. This will sterilize the leaf tissue.

3. Put on gloves and wipe the entire area where you are working with the bleach solution. It is now sterile and the leaf is sterile. Remove the leaf piece from the bleach-and-water solution and rinse the leaf four times with distilled water.

4. Lay the sterile leaf tissue onto the surface of the cooled agar mixture. Put the lid on the petri dish and place it in a room with fluorescent light or under plant lights. The room should be and remain at 75 degrees Fahrenheit.

5. Watch for shoot formation. Once there are three or four shoots, sterilize the tweezers and remove the plantlet. Dip the base of it into a rooting hormone powder and blow off any excess. Be precise and gentle when handling the plantlet.

6. Fill a pot with peat and plant the plantlet in the medium. Keep it misted with water lightly, in a warm room with moderate light. The rooting hormone will encourage the formation of roots.

References

• Aggie Horticulture: Biotechniques in Plant Breeding
• Iowa State University; Plant Micropropagation Using African Violet Leaves; Loren C. Stephens
• IGem -- Newcastle University: Making Agar Plates