The parent species of edible fungi are mainly obtained by artificial selection, mutation breeding, cross breeding and protoplasm fusion. As a general seed production professional household, the mother seed can be isolated and cultivated manually. The specific steps and operation methods are as follows.
1. Collection of provenances From the wild-cultivated cultivated population, select representative elite mushrooms are used as provenances. The standard of the mushroom is: Eight mature, positive flowering, fleshy, no pests and diseases. Collect 1-2 seedlings that meet the above criteria and number them as a material for the separation of mothers. Collected from the cultivation room should be marked with the original strain code.
2, medium preparation Agar medium is also called PDA medium, commonly used formula: 200-250 grams of potato, agar 15-20 grams, 20-25 grams of glucose, 1000 ml of water. Magnesium sulfate can also be added 1-1.5 grams, traces of vitamin B1, potassium dihydrogen phosphate 2-3 grams. The potatoes were washed and peeled, cut into thin slices, placed in an aluminum pan and boiled for 30 minutes. After being picked up, they were filtered with 4 layers of gauze and the juice was taken. The agar was then added to the juice and stirred while heating to allow the agar to fully dissolve. The glucose was then added. After a few minutes of boiling, the same four-layer gauze was used to filter the juice. The juice was taken and the juice was poured hot into a glass test tube. One-fifth of the length of the tube is installed and the mouth of the tube is tightly packed with cotton, or the juice is placed in a glass flask and the volume is 20 ml. Then placed in a pressure cooker and sterilized at a pressure of 98-108 kPa/cm for about 30 minutes. After being sterilized, they are immediately removed and the tube is slanted on the table while it is hot. After cooling, it becomes a solid slant medium.
3. There are three methods for the separation of parent species: spore ejection, tissue separation and intra-basic separation. 1 spore ejection method. The mushroom was sterilized on the surface and the water was sucked dry. The mushroom body was hung in a flask containing agar medium, so that the gown of the mushroom body was naturally scattered on the medium to germinate the mycelium. You can also cut a small mushroom body, attached to the surface of the test tube slant medium, so that spores scattered in the medium on the germination of mycelium, that can obtain the mother species; 2 tissue separation method. The sterilized mushroom is unwound by hand from the handle of the mushroom in the inoculation box, or cut with a blade to make the mushroom body form a folio. At the junction of the cap and stipe or at the pleats, a small piece of mushroom was cut with an inoculum knife, then cut into small pieces of 510 mm, and a thin piece was picked up with an inoculation needle to access the center of the slant medium. Tubes were inoculated with a small piece of thin strips. After the germination of mycelium, the mother species could be obtained. Select the wood section of the long mushroom, cut the bark and the xylem of the surface, sterilize with 70% alcohol, saw into 1cm thick slices, put 0.1% mercury in mercury disinfection for 1-2 minutes, and then sterilize Wash off the residue. Then, the small slices are cut into 0.5-1 cm wide strips, which are inserted into the center of the slant medium. After the hyphae are grown, the mother seed is obtained. It is also possible to sterilize the mycelium in the bag with long-lasting mushrooms and then use the inoculation needle to take the mycelia with pure color and growing vigorously in the bag and inoculate in the center of the slant medium of the test tube. After the mycelium is germinated, the same bacteria are obtained. Species.
4. Appropriate temperature culture After the strains are isolated and inoculated into the test tube by the above different methods, the test tubes shall be promptly transferred into a sterilized incubator or culture chamber, and the temperature shall be controlled at about 25° C. so that the isolated spores or mycelium are suitable. Developed under temperature. The spores germinated into colonies after 3-4 days of general spore ejection, and the mycelium overgrown after 10 days; the mycelium germinated and spread on the culture medium 2 to 3 days after tissue isolation and inoculation; The mycelium resumed growth.
5. Breeding and purification The mycelium obtained by the above methods may not necessarily be of high quality, but also require breeding and purification. Therefore, after the mycelium germination, we must carefully observe and select the hyphae with pure color, robustness, normal growth, and without interruption, hook the hyphae together with the culture medium in the inoculation box, and insert the additional test tube culture medium. At 23-25 ​​°C under constant temperature conditions, culturing 7-10 days, until the mycelium overgrowth tube, and then observed, selected from the best, that is, "mother" mother.
6, transfer tube to expand the parent mother can be transferred to expand the "daughter" mother. Use the same slant medium, each can be expanded 30-50 offspring mother. Most of the production supplies are offspring mothers. It can be changed again. Generally, each branch can be expanded to 20 to 25 offsprings, but the number of tubes can not exceed 5 times.
7. The mother of the separation and breeding of the fruiting experiment must also undergo the mushrooming test. The method is: the mother seed is inoculated in a bottle or bagged wood chips culture medium, according to the temperature requirements of various species of ear species temperature, the appropriate temperature culture, until the mushroom is confirmed to be used for production.
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