f1
1. Bean juice and bean sprouts mixed with agar medium: A mixture (batter) of this medium is sterilized for 1 hour. This media is ready to be used as a pure culture (tissue culture) after smeared or planted incision (network) mature mushroom fruit body.2. Grated onion and sweet potato: Grated raw material mix flour noodles in palm (palm) and put in a solution to the composition: potato 100 grams; onions 50 grams; flour 150 grams sugar palm, and to 150 grams.3. Potato Dextrose Yeast Extract Agar (PDY): The composition of the mushroom growing medium noodles have been successfully used in breeding F1 mycelium in the Central Seed Parent Ngipiksari, Yogyakarta. The composition of this medium consists of potato dextrose (glucose), and flour in order.Preparation of growth media PDY starting from leaching and boiling potatoes. A total of 200 grams of cleaned fresh potatoes (not peeled) and washed with clean water and then sliced (chopped) and then washed again repeatedly until the used wash water was clear. Once clean, thinly-sliced potatoes rinsed again with distilled water (distilled water). The trick, potato slices soaked in a panel for 10 minutes, then boiled in 700-1000 ml of water (distilled water) for I hour so that the water is shrinking lived 500-600 mi. Then, the water decoction (extract) is filtered with flannel or other cloth or filter in the eyes small and filtered water is collected in a bottle.Add a few milliliters of water in the extract (water boiled potatoes) that have been filtered so that its volume reached 1,000 ml. Tam also even 9-15 grams of flour in order and 10-20 grams of glucose (dextrose) and then stirred and boiled in an autoclave for 15 minutes at a pressure of 15 lbs.Done done directly boiling cooling. Growth media that has been cold can be immediately included in the test tube culture. Each 1 (one) liter of artificial growth media can be used as a medium for growing pure cultures (tissue culture) oyster mushrooms as much as 150-200 tube culture. Preferably, this artificial growth media used immediately so that ga 卢 not contaminated by pollutants (pollutan), bacteria, or micro-organisms (microorganisms) other destructive and malign artificial growth media.If the amount of artificial growth media prepared exceeds the capacity of a test tube culture, then the rest of the growing media should be stored in a cold and sterile room temperature. The remaining artificial media can be used for breeding the next period.The next step is to enter the growing medium in a test tube as much as I spoon, then plugged with cotton. Blockages cotton outside the test tube wrapped with paper baking sheet and tied with a rope Keret (see Figure 5). Repeat a similar job for breeding F, others.Furthermore, the reaction tubes and the contents included in an autoclave or sterilizer automated to be done at a temperature sterilization
125'C for 1 hour. To save at the same time streamline the sterilizer, the positions of reaction tubes arranged in a row it tilted to one side or oblique cross (see Figure 6).Completion of sterilization, the test tubes are left for several hours until the temperature is cold. Then, test tubes containing sterile growth media is inserted into the sterile room anyway. Remove the baking paper cotton cover and store the tube in special racks (storage racks) in a tilted position. Tujuannnya is that the media spread the fungus grows on the walls of the reaction tube dalarn at once so spread the growth of mushroom mycelium in the test tube so as to facilitate the implementation of the decision to the next stage of breeding. Test tube is left for 24 hours so that sterile growing medium to cool at room temperature.The next step is to prepare the implementation of tissue culture, the inoculation (planting seedlings) in the form of incision (the network) mushroom fruiting bodies containing mature basidiospora. The incision is taken from the ear mushrooms adult (age 3-4 weeks since the formation of candidates for fungal or pin head) that has a large fruiting bodies, thick, and healthy.Mushroom fruit body tissue to be taken first cleaned and washed or immersed in alcohol 70 `% during 1-5 minutes. The chemicals are commonly used for washing mushrooms among other seeds alcohol 70%, 5% formalin, mercurochloride 0.001%, silver nitrate 0.1%, 0.1% mercuric cyanide, sodium or calcium hipochloride hipochloride 0.35%, carbonic acid 1%, 2% potassium permanganate, and hydrogen peroxide 3%.Mushroom fruit body is clean and sterile is placed on a board or other sterile container, then placed on the table culture. Breeding table is activated, the lights turned on, and the suction machine (filter) air is turned on by pressing the button (knob) pengontak.Half an hour since breeding diaktitkan table, then all the test tubes containing sterile growing medium which has cold storage and shelf removed and placed on a table of breeding. Later. cotton penyumbatnya opened.Mushroom fruiting body parts are the thickest lies in the "armpit" it (see Figure 7). In this section there are sources
branching hyphae or mycelium or basidiospora bag. This section is cut as wide as 0.1 cm, 0.1 cm thick, and the length is about 1 cm. To facilitate penyayatan, we can use a spatula (taper stemmed knife) or a sharp scalpel and sterile.Subsequently, an incision (network) fruiting bodies inserted into a test tube and the surface of the tube plugged with cotton back. Planting fruit body incision should be carried out on the table culture. Then, test tubes filled incision closed fruiting bodies are placed in storage racks in a sterile chamber (chamber cultures) and similar work is repeated for the manufacture of seed F, on the other tube which has been prepared.Any fungal fruiting bodies can be taken as many as 10-15 spores contained incision (basioiiaspora)Mushroom spores stored in a sterile room rather dark for 20 days until the threads of mycelium grown white meet growing medium. Furthermore, miselum culture was used as seed in the second stage of breeding. Test tube culture that failed and did not grow mycelium immediately discarded so as not to pollute (contaminate) a tube that grows good breeding. Mycelium is damaged can be identified from the media stinks and blackish brown. Test tube is cleaned for use in breeding (inoculation) period (days) later.
