Microclone Tissue Culture
Seed Starter Kit-
The Seed Starter kit provides seeds with the greatest possible advantages for germination. The kit should be ideal for those attempting to germinate old seeds, valuable seeds, and difficult to germinate seeds.
Tissue culture provides a microbe-free environment containing sugar, nutrients, and vitamins that provide developing plants with energy and mineral nutrition. This kit will decontaminate the surface of seeds, hydrate them with a solution containing sugar and a mild nutrient/vitamin solution, and germinate them on a sterile medium. When plants are actively growing, they are inoculated with beneficial microorganisms and planted in larger containers and raised as normal.
Seeds can be cleaned and added in batches such as when mixing varieties or doing a trial run. We recommend that with any new technique, the user should dry run to make sure all the parts are nearby when they are needed and that reaching them is fast and convenient. And of course it will be unfamiliar the first time through so maybe washing six healthy young seeds will give good practice and demonstrate how the cleaning and growing stages will go. Just make sure to leave enough of the solutions for both batches.
The materials in the kit are:
1) Seed wash vial with thermometer and drainage holes
2) Seed surface sterilizing solution powder (A)
3) Seed hydrating solution (B)
4) Sterile seed germination media-ten tubes and tray
5) Forceps 8”
6) Sealing tape
7) Two tubes of tissue culture medium for seed rescue
First, wash hands before using kit. Hydrate Seed Sterilizing solution (A) in 60ml water, about ½ full in Wash Vial. Find location that will maintain at 26 degrees C. Good warm locations are bright indirectly lighted windows, above the refrigerator, on top of a nightlight or similar. Use cardboard as an insulating spacer if needed. Clean seeds of any surface debris and place in wash vial. Swirl occasionally and roll in vial to wash seed surface and inside of vial. You may want to cover holes with small pieces of tape but covering with clean fingers is OK. Soak seeds for one hour.
The Sterilizing Solution may be saved in a separate container for using again within a few days. Drain Sterilizing Solution and add Hydrating Solution (B) and soak in warm location until seeds sink. The root radical may not yet appear. Usually about 24 hours. The ideal temperature for soaking is 26-28 degrees C. It is a good idea to test your warm location before using real seeds.
Seed planting location setup a clean handling area:
Seeds can be planted in tubes on any clean surface, however, a clean hood helps prevent contaminating organisms from getting into the sterile planting tubes when planting. It is simply a plastic storage container turned on its side to prevent air from drafting over the open containers.(see picture) Set up in a clean kitchen or bathroom, if possible. HEPA air filtering is always recommended in any growing environment and especially clean handling environment.
Clean forceps with an alcohol spray before using. 70% or 91% is fine. Wash and spray hands for added protection. Spray inside area of forceps too. A tall narrow tube of alcohol is a good place to clean tools (see setup in the picture). Forceps are used to pick up and plant seeds. Have the planting tubes within reach and work quickly.
Drain sugar solution into a small cup in the clean hood. Loosen the lid of one of the planting tubes. Pick up your first seed , using the forceps, with the root end pointed away from the forceps. The point on the seed end should be like the pointy end of a spear, the best way to handle and plant a decontaminated seed. Stab into media about ¼” deep along the edge of the tube. The progress of the germination can be observed through the plastic.
Grow the tube plants under a small fluorescent light in a cool area not exceeding 76 degrees. The closed tube is like a greenhouse and will be a few degrees warmer inside the tube. Provide ten watts per sq foot of light, same as clone lighting.
When the seedling stands up and the leaves are open to the sky, the seedling is ready to be planted. Use the forceps to remove it along with attached soil and plant in a small pot for continued growth under the fluorescent light. Add root microbes if you have them. Cover with a dome if dry air is a concern. Seedlings adapt to new soil within a few days and will grow as normal.
Tissue Culture Seed Rescue: The kit includes two tubes of tissue culture medium just in case seeds do not germinate completely. You may also use these tubes to introduce your plant into tissue culture and multiply indefinitely using a Microclone Plant Tissue Culture Kit.
I f seeds in sterile media only partially germinate, follow the sterile technique used for planting in sterile soil to clean forceps and move seedling to TC media. Seedling can be rinsed in leftover Sterilizing Solution (A) before planting in TC gel. The sugar and nutrients in the TC tube will feed the living cells. TC seedlings will either stand up as expected and can therefore be planted in soil, or must be maintained in tissue culture environments until viable shoots can be produced.
— Ships from DL: Livermore, California USA.