Separation and purification method for Russula depalleus (Pers.) Fr. spawns

A technology for separation and purification of mycelium, applied in the field of wild mushroom mycelium separation and purification, can solve the problems of low success rate of mycelium separation, single, unfavorable mycelium growth, etc., and achieve the effect of easy operation, simple process and convenient operation

Inactive Publication Date: 2019-02-19
ZHEJIANG OCEAN UNIV
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  • Abstract
  • Description
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Problems solved by technology

[0004] The prior art, such as the Chinese invention patent with the authorized announcement number CN 105219652B, discloses a method for separating and purifying the hyphae of wild Bachu mushrooms. First, the tissues or ejected spores of wild Bachu mushrooms are cultivated on PDA medium for 6-8 days. The heterozygous hyphae were obtained in 1 day; then the heterozygous mycelium was cultured on a special medium for 7-10 days to obtain a single homozygous filamentous mycelium; the homozygous mycelium was picked and transferred to a PDA medium for culture 6-8 days; however, the general PDA medium is used in the early stage of the invention, which is not conducive to the growth of mycelium, and the success rate of mycelium separation is low

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  • Separation and purification method for Russula depalleus (Pers.) Fr. spawns

Examples

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Embodiment 1

[0036] A method for separating and purifying purple-black russula mycelia, specifically comprising the following steps:

[0037] (1) Disinfection of wild purple and black russula strains

[0038]Disinfect the tissues or spores of wild Russula russula; sterilize the tissues of wild Russula with 45% alcohol solution ultrasonically, the ultrasonic temperature is 21°C, the power is 32W, and the action time is 6min; then under sterile conditions, Cut longitudinally and tear apart the stipe or the middle part of the cap with a surgical blade that has been burnt by flame, and pick a tissue block with a length of 0.4 cm and a width of 0.2 cm at the junction of the cap and the stipe; carry out ultraviolet disinfection on the spores : Ultraviolet rays with a wavelength of 280nm are used to sterilize for 12s at a temperature of 21°C and an ambient relative humidity of 60%; wild purple and black red mushroom tissues are selected from the joints of the stipe and the cap. It is easy to dis...

Embodiment 2

[0053] A method for separating and purifying purple-black russula mycelia, specifically comprising the following steps:

[0054] (1) Disinfection of wild purple and black russula strains

[0055] Disinfect the tissues or spores of wild Russula russula; sterilize the tissues of wild Russula with 50% alcohol solution ultrasonically, the ultrasonic temperature is 22°C, the power is 35W, and the action time is 7min; then under sterile conditions, Use a flame-burned scalpel to cut longitudinally and tear apart the middle part of the stipe or cap, and pick a tissue block with a length of 0.5 cm and a width of 0.3 cm at the junction of the cap and the stipe; carry out ultraviolet disinfection on the spores : Ultraviolet rays with a wavelength of 300nm are used to sterilize for 15s at a temperature of 22°C and a relative humidity of 65%;

[0056] Wipe the surface of the separated tissue block with 70% alcohol cotton ball, and wrap the tissue with alcohol cotton for 2 minutes; make th...

Embodiment 3

[0070] The optimal conditions for the mycelium growth of purple-black russula mushroom were determined by orthogonal experiment method, which was determined by measuring the growth rate and growth amount of the mycelia;

[0071] Wherein, the method for measuring the growth rate of mycelia is: place the culture medium plate in step 6 on the microscope stage, check the eyepiece micrometer of the microscope used at the same time, and calculate the length of each grid; select the top of a single mycelium at the edge of the colony Focus under a low magnification lens; then put the eyepiece micrometer parallel to the hyphae, and select the part where the hyphae begins to appear side branches coincides with a line on the eyepiece micrometer, this point is the "reference point"; Measure the length of mycelium growth once in a certain period of time. After several observations, the growth rate of mycelium can be calculated according to the time and the length of the top extension;

[0...

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Abstract

The invention provides a separation and purification method for Russula depalleus (Pers.) Fr. spawns, and belongs to the field of separation and purification of wild mushroom spawns. The separation and purification method comprises the steps of disinfection of Russula depalleus (Pers.) Fr. strains, spawn separation and spawn purification, wherein wide Russula depalleus (Pers.) Fr. tissue is disinfected with an alcoholic solution under the help of ultrasonic waves, and spores are subjected to ultraviolet disinfection; during spawn separation, an improved PDA culture medium is adopted, and the PDA culture medium comprises an undergrowth extract from the wild Russula depalleus (Pers.) Fr. source area and Chinese mugwort powder. A preparation method for the undergrowth extract comprises the steps that humus soil from the wild the Russula depalleus (Pers.) Fr. source area is collected, fermented and subjected to shaking culture by using a constant-temperature shaker, the fermented culture solution is subjected to low-speed centrifugation and high-speed centrifugation in sequence, supernatant liquid is taken to be filtered and sterilized, and then storage is conducted for standby. The wild Russula depalleus (Pers.) Fr. spawns are separated and purified for the first time, operation is easy, the spawn separation success rate is high, the spawn purification rate is high, and purification is precise.

Description

technical field [0001] The invention belongs to the field of separation and purification of mycelia of wild mushrooms, and in particular relates to a method for separation and purification of mycelia of purple-black russula mushrooms. Background technique [0002] Purple and black Russula (Latin scientific name: Russula depalleus (Pers.) Fr.) Agaricaceae, Russulaceae, Russula are economic fungi. It grows solitary, scattered or in groups in coniferous forests or mixed forests in summer and autumn. It is edible and tastes good. This fungus is an ectomycorrhizal fungus of trees. [0003] Purple-black Russula, a wild fungus, has not been successfully domesticated so far. Although it is widely distributed, its quantity is rare. With people's understanding of the nutritional and health effects of edible fungi, the demand is increasing day by day. In particular, the market price of wild edible fungi has increased significantly, and has decreased year by year due to the extinct c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N13/00C12R1/645
CPCC12N1/14C12N13/00
Inventor 崔大练马玉心
Owner ZHEJIANG OCEAN UNIV
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