Flammulina velutiper suitable for industrial cultivation and molecular identification method of Flammulina velutiper
The technology of Flammulina velutipes and Wanjin is applied in the field of Flammulina velutipes Nongwanjin No. 8 and its molecular identification.
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Embodiment 1
[0015] Embodiment 1: Select Flammulina velutipes 'F0321' and 'F0323' that are mainly planted as hybrid parents, select 20 single-spore strains of parent 'F0321', and 20 single-spore strains of 'F0323' grow faster, and the aerial hyphae are more developed The monospore strains were paired according to the full combination method, and a total of 60 combinations were prepared, and the number of hybrids containing lock joints was 47 after microscopic examination. After the initial fruiting test, 20 hybrids capable of fruiting were selected and entered. Double screen. After re-screening, 14 hybrids produced fruit normally, and 13 hybrids had yields higher than the control strain 'Nongjin No. 3', but only 3 strains had yields higher than 400g / bottle, of which Flv0012 The comprehensive traits are good, and the expected breeding goal is achieved. The hybrid strain is named "Nongwanjin No. 8" and will be used for subsequent batch tests.
[0016] The differences between the 'Nongwanjin...
Embodiment 2
[0022] Example 2: Using SV detection technology to compare the DNA differences between this strain and its parent.
[0023] Test materials: 5 strains of Flammulina velutipes, among which the strain to be tested is 'Nong Wan Jin No. 8', the control strains F0321, F0323, F0303 (Nong Jin No. 3), and Nong Jin No. 6.
[0024]Inspection method: According to the genome sequencing data, Kmer subtractive assembly technology is used to obtain differential fragments (SV), and several SV sequences are selected accordingly to design specific primers to amplify the samples to be tested. (For references, see Alkan, C., Coe, B. P. & Eichler, E. E. Genome structural variation discovery and genotyping. Nature reviews. Genetics, 2011 (12): 363–376; - Modernization of Traditional Chinese Medicine, 21(4): 764-774.). SV test primer sequences were synthesized by Qingke Biotechnology Co., Ltd.
[0025] The mycelium activation, cultivation, genomic DNA extraction and electrophoresis detection of the...
Embodiment 3
[0070] Example 3: 'Nongwanjin No. 8' has carried out multiple yield stability tests. The results show that 'Nongwanjin No. 8' meets the requirements of industrial cultivation, and its yield is significantly higher than that of the parent F0303 (Nongjin No. 3).
[0071] Stable production test strain: 'Nongjin 3' (Flv0303).
[0072] Test location: Fujian Wanchen Biotechnology Co., Ltd.
[0073] Test arrangement: uniform production of strains, factory cultivation at the test site according to local production management methods, bottle cultivation, and multi-zone mobile bed frame cultivation (plastic bottle, 0.43kg (dry material) / bottle).
[0074] Test results: During the experimental seed production period, the mycelium of 'Nongwanjin No. 8' was thicker in PDA, and the optimum growth temperature was 20 ℃. The cultivation results of 'Nongwanjin No.8' are as follows: figure 1 shown.
[0075] Cultivation findings: (1) The time from inoculation to harvest, 'Nongwanjin 8' and the...
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