阅读说明：本技术 一种野生毛柄钉灰包菌株的专用培养基及培养方法 (Special culture medium and culture method for wild petiole nail ash wrapped strain ) 是由 罗影 贾培松 努尔孜亚·亚力买买提 贾文婕 魏鹏 郝敬喆 宋博 高海峰 丁丽丽 关 于 2020-06-29 设计创作，主要内容包括：本发明公开了一种野生毛柄钉灰包的专用培养基及其培养方法。通过菌株分离、基础培养基、母种培养基和培养条件的筛选试验,得到野生毛柄钉灰包菌株培养的专用培养基及其培养方法。本发明所述的培养基按照质量体积比计包括：胰蛋白胨5-15、酵母粉4-6、NaCl 8-12、木屑粉20-30、燕麦粉20-30、琼脂粉15-30和蒸馏水800-1500。利用该培养基培养的毛柄钉灰包菌丝体生长速度可达0.52-0.64mm/d,菌丝长势强,本发明提供的技术方案解决了现有技术未见报道有关野生毛柄钉灰包菌株适合的专用培养基的技术问题,本发明提供的专用培养基及野生毛柄钉灰包菌株的培养方法,能够满足开展生物学研究的要求,为其人工驯化栽培方面具有现实的实用价值和应用推广价值。(The invention discloses a special culture medium for wild petiole nail ash bags and a culture method thereof. The special culture medium for culturing the wild petiole glochidion strain and the culture method thereof are obtained through strain separation, a basic culture medium, a mother culture medium and a screening test of culture conditions. The culture medium comprises the following components in percentage by mass and volume: 5-15 parts of tryptone, 4-6 parts of yeast powder, 8-12 parts of NaCl, 20-30 parts of sawdust powder, 20-30 parts of oat powder, 15-30 parts of agar powder and 800-1500 parts of distilled water. The growth speed of the ash-coated mycelium of the petiole nails cultured by using the culture medium can reach 0.52-0.64mm/d, the growth vigor of the mycelium is strong, the technical scheme provided by the invention solves the technical problem that the prior art does not report the suitable special culture medium for the wild petiole nail ash-coated strain, and the special culture medium and the culture method for the wild petiole nail ash-coated strain provided by the invention can meet the requirement of developing biological research and have practical value and application and popularization value in the aspect of artificial domestication and cultivation of the strain.)

1. The special culture medium for the wild petiole nailing ash bag is characterized by comprising the following components in percentage by mass and volume: 5-15 parts of tryptone, 4-6 parts of yeast powder, 8-12 parts of NaCl, 20-30 parts of sawdust powder, 20-30 parts of oat powder, 15-30 parts of agar powder and 1500 parts of distilled water.

2. The special culture medium for the wild petiole nailing ash bag according to claim 1, which is characterized by comprising the following components in percentage by mass and volume: tryptone 10, yeast powder 5, NaCl10, wood dust 25, oat flour 25, agar powder 20 and distilled water 1000.

3. The special culture medium for the wild peduncle tack ash bag according to claim 1, wherein the oat flour is dried in the sun and crushed before use and is sieved by a 20-mesh sieve.

4. The special culture medium for the wild stalk nail ash bag according to claim 1, wherein the wood dust powder is dried and crushed before use and sieved by a 20-mesh sieve.

5. A method for culturing a wild petiole nailing ash bag strain by adopting a special culture medium for the wild petiole nailing ash bag is characterized by comprising the following steps of:

(1) preparation of a culture medium: adding tryptone, yeast powder and NaCl into deionized water, stirring until all solutes are dissolved, adjusting the pH value to 7.0 by using NaOH solution with the molar concentration of 1.0mol/L, then adding wood dust powder, oat powder and agar powder, finally fixing the volume to 1000mL by using the deionized water, subpackaging the prepared culture medium into test tubes while hot, plugging test tube openings by using silicon rubber plugs, bundling every 10 test tubes, and using newspaper or agar bundlesPacking kraft paper tightly with one end of silica gel plug, sterilizing in autoclave under 1.0-1.5kg/cm²At the temperature of 115 ℃ and 121 ℃ for 15-30min, taking out the test tube and placing the test tube on an inclined plane, and cooling and solidifying the test tube to obtain a mother culture medium;

(2) separating entity tissues of the wild petiole nailing ash steamed stuffed bun: collecting young fruit bodies in a wild carpopodium nail ash bag distribution area, wrapping the fruit bodies with newspaper or a breathable bag, bringing the fruit bodies back to a laboratory for strain separation, cleaning impurities on the surfaces of the fruit bodies with a brush, carefully wiping the surfaces of the fruit bodies with 75% alcohol for disinfection, longitudinally breaking the fruit bodies under the aseptic condition, taking out the fungus and meat tissues of the fruit bodies with the sizes of soybean grains on the sections of the broken fresh fruit bodies with a pair of burning and sterilizing forceps or a scalpel, and inoculating the tissues to the inclined plane of the special mother culture medium obtained in the step (1);

(3) placing the inoculated strains in a constant-temperature incubator at 22-28 ℃ for dark culture, taking hypha blocks which grow fast and are free from mixed bacteria pollution to transfer to the surface of another slant culture medium again for later use after the hypha grows well, culturing at 22-28 ℃ until the hypha grows over the slant and is free from mixed bacteria, namely the wild oudemansiella pedunculata mother strain, and preparing the culture method for obtaining the wild oudemansiella pedunculata gray bag.

Technical Field

The invention relates to the technical field of culture of wild edible fungus strains, in particular to a special culture medium and a culture method for wild petiole nailing ash-coated strains.

Background

The edible fungi are generally rich in protein, polysaccharide and vitamins, have low fat content, generally have the physiological effects of improving the immunity of human bodies, improving the functions of the human bodies and treating cardiovascular diseases, and are known as healthy foods in the 21 st century. With the improvement of the living standard of people, the demand of people on novel edible fungus varieties is gradually increased, so that the development of new wild edible fungus varieties becomes the demand of situation development.

The peganum (Battarrea.) was established in 1801, with the endocapsule ring-opened at the junction with the stipe and the spore having a unique elastic filament that is distinct from other fungi, 3 of which are described worldwide. The ash-spiked bags are large-sized basidiomycetes, the appearance characteristics of the basidiomycetes are greatly changed, and partial identification characteristics are difficult to obtain sometimes, so that the conception of the species is disordered, and the ash-spiked bags with hairy stems are considered to be a form of the phalloidic ash-spiked bags at one time. Liu hong and Van Li et al consider that the petiole nail ash bag (B.stevensii) and the phalloidic nail ash bag (B.pharoidides) of the Battarea genus are two independent species.

The basil fruits are large, buried underground, spherical, edible, the coating is connected with the cap-shaped top end of the stalk, grows on the ground after being matured, and the coating is annularly cracked at the connection part with the stalk. Currently, there is no report of ash bag domestication cultivation of the wild petiole nails, and the research of the prior art shows that the wild petiole nail ash bag bacterial strain basically does not grow on a common potato glucose agar (PDA) culture medium, and the research of the prior art does not report a suitable special culture medium related to the wild petiole nail ash bag bacterial strain, so that the requirement of developing biological research is difficult to meet, and the artificial domestication cultivation research as a bacterial strain is also not convenient, so that a separate culture method for developing the wild petiole nail ash bag bacterial strain is urgently needed.

Disclosure of Invention

Aiming at the technical current situation that a culture medium suitable for wild petiole nail ash wrapped strains is not reported in the prior art, the wild petiole nail ash wrapped strains basically do not grow on a common potato glucose agar (PDA) culture medium, the requirement for carrying out biological research is difficult to meet, and artificial domestication cultivation research is also inconvenient to use as strains, the invention aims to provide the special culture medium for the wild petiole nail ash wrapped strains and the culture method for the wild petiole nail ash wrapped strains. The special culture medium suitable for the wild hirsutella nail ash wrapped strain is obtained through strain separation and screening tests of a basic culture medium, a mother culture medium and culture conditions, the growth speed of the wild hirsutella nail ash wrapped mycelium cultured by the culture medium is high, the growth vigor of the mycelium is strong, the requirement for developing biological research can be met, the artificial domestication and cultivation research on hirsutella nail ash wrapped strains is facilitated, and a technical basis is laid for the artificial domestication and cultivation of the hirsutella nail ash wrapped strain.

In order to achieve the technical purpose, the technical scheme adopted by the invention comprises the following steps:

the invention specifically provides a special culture medium for wild petiole nailing ash bags, which comprises the following components in percentage by mass and volume: 5-15 parts of tryptone, 4-6 parts of yeast powder, 8-12 parts of NaCl, 20-30 parts of sawdust powder, 20-30 parts of oat powder, 15-30 parts of agar powder and 1500 parts of distilled water.

Preferably, the special culture medium for the wild petiole nailing ash bag comprises the following components in percentage by mass and volume: tryptone 10, yeast powder 5, NaCl10, wood chip powder 25, oat powder 25, agar powder 20 and distilled water 1000.

In the invention, the oat flour is dried and crushed before use and is sieved by a 20-mesh sieve.

In the invention, the wood dust powder is dried and crushed before use and is sieved by a 20-mesh sieve.

Meanwhile, the invention provides a culture method of wild petiole nailing ash bags, which adopts the special culture medium and comprises the following steps:

(1) preparation of a culture medium: weighing each reagent according to a formula, adding tryptone, yeast powder and NaCl into deionized water, stirring until each solute is dissolved, adjusting the pH value to 7.0 by using a NaOH solution with the molar concentration of 1.0mol/L, then adding wood dust powder, oat powder and agar powder, finally fixing the volume to 1000mL by using the deionized water, subpackaging the prepared culture medium into test tubes while the culture medium is hot, wherein each test tube is 10mL, a test tube opening is plugged by using a silica gel plug, each test tube opening is bundled by 10 pieces, one end with a silica gel plug is tightly wrapped by using moistureproof paper, and the culture medium is placed into an autoclave for sterilization under the pressure of 1.0-1.5kg/cm²And at the temperature of 115 ℃ and 121 ℃ for 15-30min, taking out the test tube, placing the test tube on an inclined plane, and cooling and solidifying the test tube to obtain a mother culture medium.

(2) Collecting young fruit bodies in a wild carpopodium nail ash bag distribution area, wrapping the fruit bodies with newspaper or a breathable bag, bringing the fruit bodies back to a laboratory for strain separation, cleaning impurities on the surfaces of the fruit bodies with a brush, carefully wiping the surfaces of the fruit bodies with 75% alcohol for disinfection, longitudinally breaking the fruit bodies under the aseptic condition, taking tissues with the sizes of soybean grains on the sections of the broken fresh fruit bodies by using burning sterilized tweezers or a scalpel, and inoculating the tissues to the inclined plane of the special mother culture medium obtained in the step (1).

(3) Placing the inoculated strains in a biochemical incubator at 22-28 ℃ for dark culture, and reserving the strains for later use when mycelia grow well; and (3) transferring the hypha blocks which grow fast and are free from foreign bacteria pollution to the surface of another slant culture medium again, culturing at 22-28 ℃, and obtaining the wild petiole nail ash bag mother strain, wherein hypha grows over the slant and is free from foreign bacteria, and the wild petiole nail ash bag mother strain is prepared.

According to the technical scheme, the special culture medium for the wild petiole nail ash bag and the culture method thereof are finally formed, and the special culture medium has the following technical effects:

(1) the invention provides a special culture medium for wild rhizopus arrestus and a culture method for wild rhizopus arrestus strains, when LB is used as a basic culture medium, the growth speed of mycelium is 0.23mm/d, the growth vigor is optimal, the strain is basically not eaten on a common PDA culture medium, then LB is used as the basic culture medium, different mother culture medium formulas are set, and the growth speed of mycelium of the wild rhizopus arrestus is 0.23-0.63mm/d, wherein the growth speed of the mycelium reaches 0.52-0.64mm/d under the culture medium and culture conditions provided by the invention, and the growth vigor of the mycelium is strong.

(2) The special culture medium for the wild peduncula nail ash bag and the culture method for the wild peduncula nail ash bag strain provided by the invention overcome the technical problem that the prior art does not report the suitable special culture medium for the wild peduncula nail ash bag strain.

Drawings

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Detailed Description

The present invention will be described with reference to examples, but the present invention is not limited to the examples.

The wood chips, oat, tryptone, agar powder, yeast powder, distilled water and the like selected in the separation culture of the wild petiole nail ash bag can be purchased through public channels, and equipment and instruments adopted in the process are common equipment in the field.