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How to Grow a Plant in a Tissue Culture

Scientists who focus on studying plants are called botanists or plant biologists. As a plant biologist, it is important to have controlled conditions for your plants to grow. In such conditions, it is easily possible to find plants with mutations that can be used in further studies. For example, a mutated plant that requires additional sunlight will not grow as quickly as wild plant types. For plant biologists to study these mutations, plants are grown in a tissue culture, where their growing environment is exactly the same as the plant next to it. A tissue culture plate is a small plastic sheet, about 3 by 5 inches and up to 1 inch thick. Within the plastic sheet are small cylindrical wells that the seeds are placed in to grow. Plants grown in these plates can then be easily and accurately compared to each other to find mutations.

Latex or nitrile gloves
Laminar flow hood
95 percent ethanol
Spray bottle
Paper towel
Sterile tissue culture plate
Sterile Murashige and Skoog agar medium
Microwave
Water bath at 50 degrees centigrade
Tweezers
Sterile seeds

Instructions

- 1
  
  Sterile technique is used to prevent contamination.
  
  Create a sterile working environment by using your spray bottle to spray a fine mist of 95 percent ethanol over your countertop or laminar flow hood bench surface. While wearing your latex or nitrile gloves, use the paper towel to wipe down the surfaces you will be working on. It is important to work in a sterile area to avoid contamination of the culture tissue plate.
- 2
  Unwrap your sterile culture tissue plate in the laminar flow hood. Be sure not to touch the inside of the wells with your hands. If you do not have a laminar flow hood to work in, do not remove the lid from the culture tissue plate.
- 3
  Adjust the temperature of your Murashige and Skoog agar medium until it is 50 degrees centigrade. This can be done by microwaving the medium at 50 percent power until it is completely liquid and then placing in a 50 degree centigrade water bath.
- 4
  Remove the culture plate lid and pour the liquid medium into the well. Depending on the type of culture tissue plate you have, there may be between 12 and 96 wells. Fill the well until it is roughly 3/4 full with Murashige and Skoog agar medium.
- 5
  Wait for the Murashige and Skoog agar medium to cool in the culture tissue plate wells. Full cooling may take up to an hour. If you are working in a laminar flow hood, keep the lid off while drying. If you are working on a countertop, replace the lid after pouring the Murashige and Skoog agar medium to prevent contamination.
- 6
  Sterilize your tweezers by dipping them in the 95 percent ethanol. Allow the tweezers to air-dry completely.
- 7
  Grasp a single seed with your sterile tweezers. Place the seed in the center of the culture tissue plate well without touching the Murashige and Skoog agar medium with the tweezers.
- 8
  Replace the lid on the culture tissue plate. It is important to prevent contamination of the Murashige and Skoog agar medium by limiting contact with air.
- 9
  Place the culture tissue plate in a cool, dark room for two days.
- 10
  Move the culture tissue plate to a controlled growth chamber or an area that receives adequate daylight for plant growth.

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