The method used to separate a portion of fruiting species, known as tissue separation. Asexual reproduction or cloning techniques are, in theory, the genetic recombination strain variation obtained does not occur, it is often produced on adoption. _Select_ pollution and robust mushroom-shaped mushrooms young mushrooms (6 points mature), after picking promptly placed in a sterile bag back inoculation chamber, together with the parent species together with test tubes and other supplies into the inoculated medium tank disinfection. Aseptic inoculation chamber, first with 75% alcohol to disinfect hands and tools, and then disinfected with 0.1% mercuric chloride solution planting mushroom surface with a sterile scalpel from the middle of the cap and stipe at the longitudinal coupling open cut at the junction of the cap and the stalk growth strip pieces, were inoculated with bacteria which small pieces of meat hook, quick access to the medium in the middle slope, stoppered with cotton wool plug at 25 ℃ for 3 to 5 days you can see the white sparse slender hyphae grow tissue around 15 to 18 days the mycelium can be covered with test tubes, bacteria pollution pick healthy and vigorous mycelial tube adapter further propagation, subject to passing the test fruiting After production.
Organizational separation method is simple, easy offspring mutate, to maintain the excellent characteristics of the original strains. Organizational separation is in the best use of the strong growth of the young fruiting bodies or Gulei as a separation material, take the junction of the cap and stipe tissue separation, the best, for those inside or outside the protection of the mushroom fungus curtain class, it takes in the bacteria inoculated gill curtain to protect young life force even stronger. For some mycorrhizal fungi, the access near the base of the stalk tissue to survive.
Common separation method:
1 fruiting separation: planting mushroom to choose flowers large cap thick, short handle, Ba Jiufen mature varieties. Two mushroom cut base, in a sterile box with 0.1% mercuric chloride flooding a few minutes, then rinse and wipe dry with sterile water or wipe the cap and stipe with 75% alcohol cotton ball 2 times for surface disinfection . Inoculation, as long as the planting mushroom torn cover and Stipe in Moe or gills at the junction, a small piece of tissue picked; moved on pda receiving medium. Cultured for 3-5 days _set_ about 25 ℃ temperature, you can see a white fluffy mycelium organization, Gatling expand that to get species. Such as mushrooms, mushroom, etc. You can use this method.
2 sclerotia separation: the fruiting bodies of fungi Poria, Polyporus, Rayone so difficult to capture. The common is it stored nutrients sclerotia. With sclerotia separated species can also be obtained. Sclerotia method is to wash the surface with alcohol or after mercuric chloride, cut sclerotia, take a small piece of tissue in the middle, about the size of soybean, inoculated on pda medium slant, incubated. Should pay attention to the sclerotia are storage organs, most polysaccharide containing only a small amount of mycelium, and therefore the picked tissue bigger, if tissue is excessively small, easy separation of species.
3 streptozotocin separation: fruiting difficult to find a part, nor sclerotia can be isolated by streptozotocin. Such as Armillaria, Armillaria false. Its method of operation is to use alcohol or mercuric chloride will streptozotocin gently wipe the surface of the black skin 2-3 times, and then remove the black outer cortex (bacteria sheath), white fungus extract marrow part; bacteria with sterile scissors to cut short marrow inoculated on a medium, incubated, i.e., a bacteria species.
Streptozotocin separation should pay attention: because of streptozotocin relatively small, relatively small separation factors, the separation of bacteria easily contaminated, so strict operation.
For example:
The famous black fungus strains "Ju three", "Dongmei 1", "Xuemei 1", "Xuemei No. 3" 1, "984" is the Heilongjiang Provincial Institute Dongmei Chaihe mushroom mushroom plant By separating the organization of wild mushrooms, domesticated breeding success.
2. Mushroom tissue separation methods
0.1% of the fruiting body chosen by the first open mercury surface water or 70% alcohol after disinfection. Rinsed with sterile water and surface water wiped off with sterile gauze. When separated from the stalk of the mushroom with a sterile scalpel to cut a little, and then hand the fruiting body broke into two, at the junction of the cap and gills, 0.3 to 0.4 cm3 cut a small piece of meat bacteria, put the shift In slant culture center. As already parachute of planting mushroom, then _select_ed strains of meat at the junction of the cap and stipe. Separation mushrooms, mushroom bud with a sterile scalpel to slit a little; then gently peel the handle cap, and the cap on top of the junction of the stipe, cut into small pieces 1 to 5 mm, then the slant culture base in. If planting mushroom has been exposed to rain, water more, should be taken as gill inoculation material. After separation of the tube outside the organization cultured in incubators. Subject to the surrounding tissue sprout mycelium, and to spread the growth medium, and then picked the robust growth of the mycelium carried Gatling culture. Organizational separation method is simple, and easy to bring bacteria, easy to obtain pure strains. But for white fungus, black fungus and other gelatinous fungus; its content is extremely mycelium fruiting bodies, such as the isolation and culture with the organization, are often difficult to succeed. |
