Tremella fuciformis separation technology
Unlike other mushrooms, the production of various strains of Tremella is a two-bacterial culture, ie, regardless of the parent, seed, or planting type, in each test tube, each strainer bottle, and each planting bag (even each inoculation hole) Both of them must allow Tremella mycelia (binuclear mycelia, also known as white hairball or spore-like mycelium) and the associated fungi Aspergillus mycelia to grow in proportion (and in a coordinated manner) in the culture medium (substance), so that they can be cultivated to have practical value. , It can be used for the actual cultivation of effective bacteria. (I) Separation Techniques Treponema pallidum spores were obtained by methods of catapult ejection, tissue separation, and ear separation. 1. Spore Ejection Method This is a method that uses fresh, mature fruit bodies to automatically eject basidiospores to obtain pure strains. Fresh, white, and disease-free fresh white fungus were used as a kind of ear. Rinse the white fungus several times with sterile water, and then use sterile gauze or blotting paper to put the white fungus on the surface The moisture drains. If there is a thin layer of water on the surface of the ear, it will affect the ejection of the basidiospore of the tremella and cause contamination of the bacteria. Then cut the ear into small pieces, put it into a flask or put it in a test tube without touching the wall or the tube wall. Then use a stainless steel hook to hook the small tab and quickly hang it in the flask or test tube. Be careful not to make the ears touch the surface of the culture medium in order to prevent the contamination of the bacteria, plug the tampon, and then place it at 20-25°C. After 24 hours, the basidiospores ejected from the ears fall in a smooth culture. A misty spore print forms on the base surface. At this time, in the sterile box or sterile room, the ear hanging in the bottle is taken out, and then stuffed with a tampon, moved to the room temperature of 20-25 °C, after 2-3 days, the surface of the medium on the There will be many milky, pasty little colonies. This is the yeast conidia of Tremella. When colonies of yeast-like conidia appear on the culture medium, in order to prevent and reduce contamination, purification should be carried out as soon as possible, that is, picking up a few conidia of tremella species without bacteria, and transferring them into a new test tube for cultivation. No matter whether the trichospores or yeast conidia are not easy to germinate into hyphae, they are difficult to use directly in production, and they are rarely used at present. However, in order to obtain the offspring of sexual reproduction of Tremella, the basidiospores must be isolated and germinated into mononuclear hyphae, and then paired experiments should be performed. 2. The method of tissue separation is different from shiitake mushrooms and oyster mushrooms. Segregation of tremella fuciformis is difficult to operate and its practical significance is not great. Because once the gums have been gelatinized, it is not easy to grow bacteria. Silk, and the white fungus or wild white fungus, exposed to the outside world The growth time is very long, and various bacteria are bound on the ear piece. When the tissue is separated, it is not easy to wash away the attached bacteria, and it is not easy to use the medicine to sterilize the surface, and it easily leads to the loss of both the white fungus and the stripe. Only the hypertrophic ear base located between the bark and the xylem is relatively easy to separate, and a pure mycelium of the white fungus can be obtained. However, the white fungus pure hyphae isolated from the ear are often yellow or orange yellow, not pure white, and the growth rate of the mycelium is relatively slow, and the effect is not ideal. 3. The ear-separation method utilizes a segment of wood (ear ear) which is a long-eared fruit body of Tremella fuciformis for separation, and is currently the most important method for obtaining valuable Tremella species. The main technology of the ear-separation method is to eliminate the interference of the bacteria in the ear and prevent the contamination of the bacteria during the separation. The mycelia of the white fungus and the hyphae must be separated. For beginners, ear separation is not easy to master and must be practiced repeatedly until mastery of separation. The main steps of the ear separation method are as follows: (1) Ear wood selection When the first and second batches of white fungus are grown, the long ear should be the largest, the best quality (especially the slice width, color white), and the least amount of miscellaneous bacteria should be used as the separation material. The separated material is air-dried, and after the insecticide (especially enamel) is fumigated with a fumigant, it is transferred to a sterile or sterile room for separation. (2) Ear Sterilization Sterilize the ears with a 2-4 cm thick wooden wheel. Then remove the bark, ear base, and place it in the inoculation box (room). Use 0.1% Hg water or 70% of the ear surface. Alcohol rubbing sterilization. Skilled persons can also separate without surface sterilization. (3) Ears are planted and cut through the ears. Then, shave very small sawdust powder inside the wood just below the ear-base. (You can't use small pieces of wood. The smaller the inoculation size, the greater the chance of getting pure Tremella species. There is a greater chance of inclusion of a little white fungus mycelium in the ash mycelium, inoculated on the surface of the culture medium in the test tube. After the inoculation, the test tube was cultured at room temperature of 20-28°C, and mycelium began to grow around the inoculated block several days later. (4) Separation results The results of the separation are different due to different separation techniques. Roughly the following results can be obtained: Only the white fungus mycelium was obtained; only the mycelium of the ash was obtained; the mycelia of the white fungus and the hyphae of the ash were obtained at the same time, but the hyphae accounted for the absolute majority; the conidia and the mycelia of the fungus were obtained. If there is a mixture of bacteria, the result of the separation is more complicated. In order to obtain valuable trear Tremella species, the tremella mycelia and the incense hyphae were isolated separately in the 3rd to 7th days after isolation. Tremella fuciformis Features: White, yellowish-white, and various intermediate colors from white to yellow, aerial hyphae erect, obliquely or flatly affixed to the surface of the culture medium, and the base mycelium is produced in the medium. The diameter of the mycelium is 1. 5-3 μm. There is a diaphragm, and there is a distinct (but not every diaphragm) interlocking joint that grows at a slower rate than ordinary mushrooms. In some cases, there will be entangled mycelium clusters on the surface of the culture medium, which will gradually gelatinize and become small primordial and grow small ears. When the double-nuclear hyphae begin to gelatinize, several burden-like cells can be found on the top of the mycelium, which means that the double-nuclei hyphae have entered the solid phase. After the secondary generation (transplantation) of the double-nuclear hyphae, new mycelium grows, or rapid gelatinization occurs, and the ear grows. It can also become slimy yeast-like conidia (depending on the strain's age and development Degree, water layer on the surface of the medium, heat or cold of the inoculation needle, light or heavy gestures during inoculation). The "yeast-like conidia" broken by the double-nuclear hyphae becomes a tremella mycelia under suitable conditions. Characteristics of the incense hyphae hyphae: The hyphae are initially colorless, and when they grow in large numbers, they are white, feathery, and often have a particularly slender stem and laterally slightly "feather-like" branches. The old mycelium turns pale yellow and pale brown. The medium gradually changed from pale brown to black or blackish green. The aerial mycelium on the surface of the culture medium is grayish white, finely velvet-like, sometimes black-stained with carbon, conidial stems nearly broom-like, and rare conidia. If woodchips are used, near-elliptic conidia from yellow-green to grass green can occur after bottle breakage. Flat black sub-seats can be produced on the segment wood surface. It is known that E. coli may not be a single species and may belong to the genus Carboniferous and the genus Carbonia. There are 3-4 species, one of which was initially identified as Hypoxylon archeri. Dongguan Anmeixuan Electronic Technology Co., Ltd , https://www.hyashine.com
In order to ensure the quality of the strains used in Tremella production, researchers first put the Tremella fuciformis hyphae "white hair group", yeast-like conidia "spore", incense hyphae (also called feathery hyphae, ear bud mycelium). ) Separately cultured separately and kept separately. Only in the season of seed production can the two pure strains be paired together and mixed and cultured to produce the parental, primary, or planting species at various levels for use by the local strain farms or growers. Therefore, it can be known that the Tremella species deposited by the research unit and the strain used by the production unit are different. The Tremella species used in the research can be said to be a kind of ancestral species. It is the ancestor of all production species in the society. Breeders call it the "original species" and are pure cultures of a single species. These pure strains are used for breeding and research purposes and are generally not supplied externally. The Tremella parent species, original species and planting species used for social production are double-bacteria cultures, ie, the mycelia or spores of Tremella fuciformis and Coprinus asperata (Ascomycetes) are inoculated simultaneously in the culture medium for mixed culture. This species is generally used only once. Otherwise, it is easy to artificially separate the two bacteria in the tube, so that the mycelia of the cultured white fungus are gradually reduced, and most of the remaining hyphae are caused by the failure of the white fungus. The following describes the separation, preparation and preservation techniques of Tremella species: