Fermentation filtrate of fir endophytic fungus and its extract, preparation method and application
A technology of endophytic fungi and fir, applied in biochemical equipment and methods, microbial-based methods, botanical equipment and methods, etc., can solve the problems of lack of research, reduce pathogenicity, reduce appressoria formation rate, The effect of long-lasting control effect
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Embodiment 1
[0026] Embodiment 1: Chinese fir endophyte Epicoccum dendrobii SMEL isolation and identification
[0027] In this experiment, healthy Chinese fir branches were collected in Nanjing Forestry University in April 2017 (collector: Huang Lin;
[0028] Contact number: 025-85427301), wash with sterile water, and then sterilize the surface with 75% absolute ethanol, remove the epidermis, transfer the internal tissue to a PDA plate, culture at 28°C, and use sterile picks after 7 days. The edge part of the colony was picked with a needle and inoculated on a new PDA plate to purify the strain to obtain the endophytic fungus strain SMEL of Chinese fir.
[0029] endophytic fungi Epicoccum dendrobii The biological characteristics of the strain SMEL are as follows: The edges of the colony of the SMEL strain on the PDA medium are neat, and the aerial mycelium is felt to flocculent, flat, white to light brown; the back of the colony is light red, and the center is light brown to brown. T...
Embodiment 2
[0031] Embodiment 2: SMEL fermentation filtrate is to the inhibitory action of Chinese fir anthracnose spore germination and appressorium formation
[0032] Preparation of mycelial blocks: Inoculate C. fir anthracnose and SMEL on PDA plates respectively, culture them at 25°C for 5 days, and use a sterile puncher to prepare bacterial blocks with a diameter of 5 mm (ie, mycelial blocks).
[0033] In order to prepare the spores of the fungus Anthracnose fir (provided by the Forest Pathology Laboratory of Nanjing Forestry University, the strain name is SMCG1#C, by 2017. Observation of the nuclear behavior of the genetic transformation of the fungus anthracnose and appressorium development. Nanjing Forestry University Nanjing Forestry University Journal (Natural Science Edition), 41(6):68-72. Open) mycelium block was inoculated into 100ml CMC liquid medium, 25°C, 200rpm shaking culture, spores were collected after 2 days and the concentration was adjusted to 1×10 5 spores / ml, and ...
Embodiment 3
[0035] Embodiment 3: the inhibitory effect of the ethyl acetate extract of SMEL fermentation filtrate to the growth of Chinese fir anthracnose
[0036] Inoculate 10 pieces of SMEL mycelium with a diameter of 5mm into a 500ml Erlenmeyer flask containing 200ml of sterile PD medium, place in a darkroom shaker at 25°C, shake at 130rpm for 40d, and collect the fermentation filtrate by filtering through a funnel; The fermentation filtrate is filtered through a 0.22-μm filter to obtain the aseptic fermentation filtrate of SMEL (after filtration, the fermentation filtrate does not contain SMEL bacteria, and the components metabolized into the fermentation liquid during the SMEL cultivation process play an antibacterial role); to the sterile fermentation filtrate Add 3 times the volume of ethyl acetate and mix thoroughly (that is, the volume of ethyl acetate is 3 times the volume of the fermentation filtrate), and then let it stand for 24 hours; use a separatory funnel to collect the up...
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