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Seed Surface Sterilization – Wash the dried seeds first under regular tap water for 30 minutes for seed surface sterilization. Then, surface sterilize the seeds with 40% Clorox and a few drops of tween-20. Shake the seeds at 80 rpm in an orbital container for 20 minutes. Pre-Germination Treatment – This step will help break the seed dormancy and maximize seed germination. All you have to do is follow this procedure to treat seeds: First, scarify the seeds in 30% sulphuric acid for around 15 minutes and then rinse them with distilled water for another 10 minutes to remove the traces of acid solution. Now, surface sterilize them again with 40% Clorox for 20 minutes and re-wash them with distilled water three times for one minute each. Now, culture the seeds in an MS medium with 30 grams of L-1 sucrose, and 2.78 grams of L-1 Gelrite without any plant growth regulator. Adjust the pH to 5.7, add agar and ten autoclaves at 121 degrees Celsius for 20 minutes, and incubate the culture at 25 degrees with 16 hours of photoperiod through cool white fluorescent lights.
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After a week, the cover can be gradually removed and the plants acclimated to stronger light and drier atmospheric conditions. You now have a collection of plants in your classroom that are genetically exactly the same. You could use these plants to carry out other experiments knowing that one common source of variation in the experiment has been eliminated. Some of these tests could include looking at plant responses to low light levels, to drought, or to saline soil conditions.
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Each container can be used to prepare about 30 tubes as above. The first container should have BAP added at the rate of 2.0 mg/l. The second container should have the NAA hormone added at the rate of 0.1 mg/L. To do this, it is necessary to make concentrated solutions of both BAP (2.0 mg/ml) and NAA (1.0 mg/ml). Add 1 ml of the concentrated BAP stock or 100 µl of the NAA concentrated stock to each 1 liter of medium that you prepare. If you use rooting hormone purchased from your local hardware or nursery supply store instead of NAA, then just follow the directions before adding to your medium.
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Sugar uptake in plant tissue cultures appears to be partially through passive permeation and partially through active transport. Sucrose also supports the maintenance of osmotic potential (osmoticum) and the conservation of water in cells. Hence, in anther culture a higher concentration of sucrose (6–12%) is used. It has been also proven that plant tissue cultures do not fix enough CO2 to sustain growth in the absence of sucrose, mainly due to limited CO2 inside the vessel.
Begonia produces one of the smallest types of seeds in the world. Miniature seed resemble dust. One ounce of begonia seed is enough for the production of 3 million seedlings. Seed starts to germinate 2 or 3 weeks after planting. Begonia can be propagated via seed, leaf- and stem-cuttings or via tuberous root. All species of begonia are divided in three major groups: tuberous, semperflorens, and the uncommon perennials. Tuberous begonias produce beautiful flowers, but they undergo period of dormancy during the winter when their foliage and flowers wilt and die.