A new strain of oosporus Ouderia microbes and its artificial cultivation method and application
A technology of artificial cultivation and bacterial strains, which is applied in mushroom cultivation, cultivation, plant cultivation, etc., can solve the problems of unstable quality, long growth cycle, and low yield, and achieve rich nutrients, short growth cycle, and high cultivation yield. Effect
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Embodiment 1
[0060] On May 15, 2016, Hu Huiping, Su Guanlong and others conducted large-scale fungal resource collection and investigation in Meihua Mountain, Fujian, and collected a specimen of Ordella genus on humus, such as figure 1 and figure 2 and the original strain was obtained by tissue isolation, named Oudemansiella raphanipes HMGIM-W160136, which was deposited in the Guangdong Provincial Microbial Culture Collection Center on April 26, 2020, and the address is: Xianlie Middle Road, Guangzhou 5th Floor, Building 59, No. 100 Courtyard, the preservation number is GDMCC NO: 61004.
Embodiment 2
[0062] The fruit bodies collected in the wild were sampled to obtain a pure PDA culture, and the pure culture was cultured with a serosol membrane-PDA medium to obtain fresh mycelia, dried at a low temperature (40° C.), ground with liquid nitrogen, and used. Ezup column-type fungal genome DNA extraction kit (Sangon Bioengineering (Shanghai) Co., Ltd.) was used to extract DNA genome, and the obtained DNA solution (DNA template) was refrigerated at -20°C for later use.
[0063] The ITS-PCR experiment was carried out with the universal primers ITS1 / ITS4 (ITS1: TCCGTAGGTGAACCTGCGG, ITS4: TCCTCCGCTTATTGATATGC, synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.) of the fungal ribosomal intergenic region, and the amplification was carried out on a Biometra PCR machine. The composition of the reaction solution (50 μl in total) is as follows:
[0064]
[0065] ITS-PCR reaction conditions were: 94°C for 5 min; 94°C for 1 min, 55°C for 1 min, 72°C for 1 min, 30 cycles; 72°C for...
Embodiment 3
[0068] Example 3 Artificial cultivation
[0069] The artificial domestication of Oudemansiella raphanipes HMGIM-W160136 collected in the field was carried out.
[0070] Taking Oudder oosporum HMGIM-W150719, HMGIM-MC-OR-0001, HMGIM-MC-OR-0002 and HMGIM-I170004 as controls, the growth period survey results of different strains are shown in Table 1, and the yield results of different strains are shown in Table 1. shown in Table 2.
[0071] 1. Culture medium (by weight percentage)
[0072] 1. Tissue isolation medium (comprehensive PDA):
[0073] Potato 200g / L, glucose 20g / L, agar 20g / L, potassium dihydrogen phosphate 3g / L, magnesium sulfate 1.5g / L, VB 10.01g / L.
[0074] 2. Mother seed medium (Red Bengal medium):
[0075] Peptone 5g / L, glucose 10g / L, potassium dihydrogen phosphate 1g / L, magnesium sulfate (MgSO 4 . 7H 2 O) 0.5g / L, agar 20g / L, 1 / 3000 red Bengal solution 100mL, chloramphenicol 0.1g / L.
[0076] 3. Production of parent seed medium (enriched with comprehensive PD...
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