Mulberry phellinus igniarius strain and cultivation method thereof
A cultivation method and technology of Phellinus Phellinus mycelium are applied to the mulberry Phellinus species and the cultivation field thereof, and can solve the problems of unstable fruit body yield and quality, difficulty in realizing large-scale artificial cultivation, and difficulty in artificial domestication of Phellinus mulberry. , to achieve the effect of small fluctuation of output and quality, high medicinal value and low management cost
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Embodiment 1
[0086] Example 1 Morphological characteristics and taxonomic identification of Phellinus sp. strain QJF-2
[0087] (1) Morphological characteristics of Phellinus strain QJF-2: two or three layered fruiting bodies, horseshoe-shaped or fan-shaped, woody and hard, the back of the cap is dark yellow to brown, the ventral surface is yellow, and the texture is tight; on PDA medium Cultured for 14 days, the diameter of the colony is 90 mm, the texture is fluffy, yellow, and the reverse side is brown; the mycelium is light brown, multi-branched, septate, and the diameter is 2.5-4.5 μm. The morphological characteristics show that it is the authentic mulberry Phellinus.
[0088] (2) ITS identification of Phellinus strain QJF-2
[0089] Extract the DNA of Phellinus Phellinus bacterial strain QJF-2 by routine method, carry out PCR amplification with ITS1 and ITS4 as primer, wherein the primer sequence of said ITS1 and ITS4 is:
[0090] ITS1:5'--3'-TCCGTAGGTGAACCTGCGG
[0091] ITS4:5'-...
Embodiment 2
[0097] Example 2 Cultivation Experiment of Phellinus QJF-2
[0098] (1) Preparation of the original species: mix 81% sawdust (half each of coarse sawdust and fine sawdust), 15% wheat bran, 2% bean cake, 0.5% lime and white sugar, and 1% gypsum according to the percentage by weight, and fully stir after adding water Evenly, the water content is controlled at 62% to 65%, bagged, and the cultivation bag is a polypropylene bag with a size of 17cm×33cm×0.05cm, tied with a rubber band, sterilized by autoclaving, and kept at 125°C for 150min. After the bacteria bag was sterilized, it was taken out and cooled, and the Phellinus QJF-2 test tube seed was inserted under aseptic conditions, and cultured at 28°C for 40 days under dark conditions to obtain the original Phellinus QJF-2 seed.
[0099] (2) Preparation of cultivars: See step (1) for the composition and preparation method of cultivar compost, inoculate the original seed obtained in step (1) on the cultivar compost according to...
Embodiment 3
[0117] Example 3 Study on the anti-tumor effect of fruiting body of QJF-2
[0118] The QJF-2 fruiting body obtained in the embodiment of the present invention and the traditional poplar Phellinus fruiting body polysaccharide were extracted by water extraction and alcohol precipitation method, and the extraction rates were 5.6% and 7.4% respectively. The two seed body polysaccharides were dissolved in ultrapure water to a concentration of 1 mg / mL, filtered and sterilized, and then diluted to 300 μg / mL with culture medium for use.
[0119] Cell lines HepG2, J82 and T24 are stored in the laboratory. After the three kinds of cells are recovered and passaged, the cells are washed with PBS, and then trypsin is added for cell digestion. The digested cells (5000 cells / well) are inoculated in 96 wells plate, at 37°C, 5% CO 2 overnight in an incubator. After the cells adhered to the wall, the culture medium was discarded, and a drug group, a negative control group and a solvent con...
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