阅读说明：本技术 一种蛹虫草栽培接种方法 (Cordyceps militaris cultivation inoculation method ) 是由 高益槐 王�忠 王锦锋 于 2019-10-22 设计创作，主要内容包括：本发明涉及一种蛹虫草,具体涉及一种蛹虫草栽培接种方法,其包括以下依次进行的步骤：将蛹虫草的分生孢子制成悬浮液后接种到培养基中,20～25℃下恒温室内培养3～7d后,经光照刺激得到蛹虫草。所述悬浮液的制备包括以下步骤：将蛹虫草菌丝收集,加入吐温80溶液震荡均匀后,用中速滤纸过滤纯化后,葡萄糖溶液稀释得到悬浮液备用。本发明方法克服了本领域技术人员的技术偏见,将蛹虫草分生孢子制成悬浮液,并接种到培养基中,培植出无虫子体的蛹虫草。经过试验证明,成功生长出菌丝,接种第3天就生长白色菌丝,而且生长更均匀。(The invention relates to cordyceps militaris, in particular to a cordyceps militaris cultivation inoculation method, which comprises the following steps of: preparing conidium of cordyceps militaris into a suspension, inoculating the suspension into a culture medium, culturing in a constant-temperature room at the temperature of 20-25 ℃ for 3-7 days, and performing illumination stimulation to obtain cordyceps militaris. The preparation of the suspension comprises the following steps: collecting Cordyceps militaris mycelium, adding Tween 80 solution, shaking, filtering and purifying with medium-speed filter paper, and diluting with glucose solution to obtain suspension. The method overcomes the technical prejudice of technicians in the field, and the cordyceps militaris conidia are prepared into suspension and inoculated into a culture medium to cultivate the cordyceps militaris without insect daughter. Tests prove that hyphae grow successfully, white hyphae grow on the 3 rd day of inoculation, and the hyphae grow more uniformly.)

1. A cultivation inoculation method of cordyceps militaris is characterized by comprising the following steps of: preparing conidium of cordyceps militaris into a suspension, inoculating the suspension into a culture medium, culturing in a constant-temperature room at the temperature of 20-25 ℃ for 3-7 days, and performing illumination stimulation to obtain cordyceps militaris.

2. The cultivation inoculation method of cordyceps militaris as claimed in claim 1, wherein the preparation of the suspension comprises the following steps: collecting Cordyceps militaris mycelium, adding Tween 80 solution, shaking, filtering and purifying with medium-speed filter paper, and diluting with glucose solution to obtain suspension.

3. The cultivation inoculation method for cordyceps militaris as claimed in claim 2, wherein: the concentration of the Tween 80 solution is 0.01-0.05%, and the number of conidia in the Tween 80 solution is 10⁶～5×10⁷Inoculating Cordyceps militaris hypha per mL.

4. The cultivation inoculation method for cordyceps militaris as claimed in claim 2, wherein: the concentration of the glucose solution is 2-5%, and the dilution multiple is 5-10 times.

5. The cultivation inoculation method of cordyceps militaris as claimed in claim 2, wherein the preparation of conidium hyphae of cordyceps militaris comprises the following steps: and cutting the cordyceps militaris mother seeds into blocks, inoculating the blocks into a PDA slant culture medium, putting the PDA slant culture medium into an incubator at the temperature of 23-28 ℃ for dark culture for 6-8 days, and growing hyphae on the PDA slant.

6. The cultivation inoculation method of cordyceps militaris as claimed in claim 1, wherein: the culture medium is mainly prepared from the following components in parts by weight: 20-40 parts of rice, 3-6 parts of peptone, 10-30 parts of sucrose and MgSO₄1 to 3 parts, 1 to 3 parts of monopotassium phosphate and 0.5 to 1.5 parts of vitamin B1.

7. The cultivation inoculation method of cordyceps militaris as claimed in claim 1, wherein: the conidium suspension is inoculated into a culture medium according to the volume percentage of 0.5-1%.

8. The cultivation method of cordyceps militaris as claimed in claim 1, wherein the step of light stimulation comprises:

s1, after the inoculated culture medium is illuminated for 10-14 hours, the culture medium is placed in the dark for 10-14 hours;

s2 the loop of step S1 is performed for 5-7 times.

Technical Field

The invention relates to cordyceps militaris, and particularly relates to a cordyceps militaris cultivation inoculation method.

Background

In the aspect of cordyceps militaris cultivation inoculation technology, the most applied method is to inoculate liquid strains cultured by liquid, and the method has the advantages of high inoculation efficiency, quick hypha growth and low pollution. The process of the method comprises the steps of mother seed culture → liquid seed culture → inoculation culture → hypha growth → sporophore growth → harvest. Although the growth cycle of cordyceps militaris hypha is short, liquid strains are cultured firstly, so that the culture time is longer, and the production efficiency is greatly reduced in the production process.

The university of Ludong filed an invention patent of a culture method of Cordyceps militaris (patent No. CN105733957A) for effectively replacing liquid strains with slant solid strains of Cordyceps militaris, wherein the patent mentions that the solid strains are directly used for inoculation instead of liquid strains, and the method is to mix the solid strains with glucose solution and then directly inoculate, so that the method can actually shorten the period. However, the solid culture medium is adopted and is directly mixed with the glucose solution for inoculation, the slant strain and the glucose injection are required to be extruded out from a needle after being mixed, impurities are easily mixed in the mixed mode for multiple times, the inoculated strain not only comprises conidia, the production efficiency is reduced, and the degradation of cordyceps militaris can be reduced without any suggestion.

Disclosure of Invention

Technical problem to be solved

In order to solve the above problems in the prior art, the present invention provides a novel cultivation method which adopts conidium suspension for cultivation and inoculation, reduces pollutants and effectively inhibits the pupa worm.

(II) technical scheme

In order to achieve the purpose, the invention adopts the main technical scheme that:

a cultivation inoculation method of cordyceps militaris comprises the following steps of: preparing conidium of cordyceps militaris into a suspension, inoculating the suspension into a culture medium, culturing in a constant-temperature room at the temperature of 20-25 ℃ for 3-7 days, and performing illumination stimulation to obtain cordyceps militaris.

Further, the preparation of the suspension comprises the following steps: collecting Cordyceps militaris mycelium, adding Tween 80 solution, shaking, filtering and purifying with medium-speed filter paper, and diluting with glucose solution to obtain suspension.

Further, the concentration of the Tween 80 solution is 0.01-0.05%, and the number of conidia in the Tween 80 solution is 10⁶～5×10⁷Inoculating Cordyceps militaris hypha per mL.

Furthermore, the concentration of the glucose solution is 2-5%, and the dilution multiple is 5-10 times.

Further, the preparation of conidium hyphae of cordyceps militaris comprises the following steps: and cutting the cordyceps militaris mother seeds into blocks, inoculating the blocks into a PDA slant culture medium, putting the PDA slant culture medium into an incubator at the temperature of 23-28 ℃ for dark culture for 6-8 days, and growing hyphae on the PDA slant.

Further, the culture medium is mainly prepared from the following components in parts by weight: 20-40 parts of rice, 3-6 parts of peptone, 10-30 parts of sucrose and MgSO₄1 to 3 parts, 1 to 3 parts of monopotassium phosphate and 0.5 to 1.5 parts of vitamin B1.

Further, the conidium suspension is inoculated into a culture medium according to the volume percentage of 0.5-1%.

Further, the step of illuminating the stimulus comprises: s1, after the inoculated culture medium is illuminated for 10-14 hours, the culture medium is placed in the dark for 10-14 hours; s2 the loop of step S1 is performed for 5-7 times.

(III) advantageous effects

The invention has the beneficial effects that:

1. the method overcomes the technical prejudice of technicians in the field, and the cordyceps militaris conidia are prepared into suspension and inoculated into a culture medium to cultivate the cordyceps militaris without insect daughter. Tests prove that hyphae grow successfully, white hyphae grow on the 3 rd day of inoculation, and the hyphae grow more uniformly.

2. The inoculation culture method of the cordyceps militaris can effectively prevent the degeneration of the properties of the cordyceps militaris.

3. The key point of the cordyceps militaris conidium collection and suspension liquid preparation lies in the purification of the bacterial liquid, and compared with the prior art, the method disclosed by the invention is lower in pollution rate.

4. According to the invention, the conidium suspension liquid replaces liquid strains to carry out cordyceps militaris cultivation inoculation, so that the cultivation time is shortened, the cultivation cost is reduced, and the production efficiency is improved.

Detailed Description

For a better understanding of the present invention, reference will now be made in detail to the present invention by way of specific embodiments thereof.

A cultivation method of cordyceps militaris comprises the following steps of: preparing conidium of cordyceps militaris into a suspension, inoculating the suspension into a culture medium, culturing in a constant-temperature room at the temperature of 20-25 ℃ for 3-7 days, and performing temperature difference stimulation, illumination treatment and the like to obtain the cordyceps militaris.

In order to successfully prepare the conidium hypha of the cordyceps militaris into the conidium suspension with higher purity, the following steps are adopted but not limited: adding conidium hyphae of cordyceps militaris into a Tween 80 solution, shaking uniformly, filtering and purifying by using medium-speed filter paper, and diluting a glucose solution to obtain a suspension for later use.

In order to optimize the concentration of the suspension, the concentration of the Tween 80 solution is 0.01-0.05%, and the number of conidia in the Tween 80 solution is 10⁶～5×10⁷Inoculating Cordyceps militaris hypha per mL.

The concentration of the glucose solution is 2-5%, and the dilution times are 5-10 times.

Preparing conidium hyphae of cordyceps militaris: and cutting the cordyceps militaris mother seeds into blocks, inoculating the blocks into a PDA slant culture medium, putting the PDA slant culture medium into an incubator at the temperature of 23-28 ℃ for dark culture for 6-8 days, and growing hyphae on the PDA slant.

The culture medium is mainly prepared from the following components in parts by weight: 20-40 parts of rice, 3-6 parts of peptone, 10-30 parts of sucrose and MgSO₄1 to 3 parts, 1 to 3 parts of monopotassium phosphate and 0.5 to 1.5 parts of vitamin B1.

The conidium suspension is inoculated into a culture medium according to the volume percentage of 0.5-1%.

The step of light stimulation comprises: illuminating the inoculated culture medium for 10-14 h, and then placing in the dark for 10-14 h; the step is circulated for 5-7 times.

At present, the cordyceps militaris cultivation inoculation is liquid strain inoculation, and the method is key and is an indispensable key factor; the liquid spawn culture needs equipment and instruments and needs time, the steps of the invention are simplified on the original cultivation inoculation technology of the cordyceps militaris, and conidium suspension is directly used for inoculation, namely the process is as follows: mother seed culture → conidium collection (suspension) → inoculation culture → hypha growth → fruiting body growth → harvest; the method saves the link of liquid spawn, simplifies the culture steps, shortens the period, reduces the cost and improves the production efficiency under the condition of achieving the same or better culture effect. The method directly collects the conidia without repeated operation, and the operation is simpler.