CN108901620A - A kind of straw mushroom medium and preparation method thereof and cultural method - Google Patents
A kind of straw mushroom medium and preparation method thereof and cultural method Download PDFInfo
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- CN108901620A CN108901620A CN201811007192.3A CN201811007192A CN108901620A CN 108901620 A CN108901620 A CN 108901620A CN 201811007192 A CN201811007192 A CN 201811007192A CN 108901620 A CN108901620 A CN 108901620A
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- straw mushroom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
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Abstract
The invention discloses a kind of straw mushroom medium and preparation method thereof and cultural methods, are related to volvaria volvacea cultivation technology field.The straw mushroom medium contains potato water extract, glucose, peptone and dregs of beans water extract.It can make straw mushroom that good hickory chick be presented by the addition of peptone and dregs of beans water extract to grow situation, show the feature that mycelia is sturdy, density is big and the speed of growth is fast.
Description
Technical field
The present invention relates to volvaria volvacea cultivation technology field, in particular to a kind of straw mushroom medium and preparation method thereof and
Cultural method.
Background technique
Straw mushroom (Volvariella volvacea (Bull.:Fr.) Sing.) also known as Volvariella volvacen, luxuriant foot mushroom originate from Guangdong
In the Nanhua Temple in Shaoguan, 300 China Nian Qian have started artificial cultivation, are passed to countries in the world by overseas Chinese in the about thirties in 20th century,
It is a kind of important tropical and subtropical zone mushroom class, is that the third-largest culturing edible fungus, China's straw mushroom yield occupy first of the world in the world, it is main
It is distributed in South China.
For straw mushroom because of its unique flavor, nutritional ingredient is high, medicinal effects are good and has very high nutrition and edible value.
Straw mushroom is a kind of good edible mushroom.The height of protein content is usually as evaluation food value height
Important evidence, straw mushroom have the characteristics that high protein and low fat.Protein content accounts for the 2.66%-5.05% of dry weight in fresh grass mushroom,
Compared with the Vegetables of daily consumption, protein content is 2 times of potato and asparagus, 4 times of tomato and carrot, citrus
6 times.In addition, containing 18 kinds of amino acid in straw mushroom fructification, wherein the required ammonia that cannot be synthesized or convert containing 8 kinds of mankind
Base acid, and the content of this 8 kinds of essential amino acids is higher, accounts for the 38.2% of its total amino acid content.
Often edible straw mushroom is with health role to human body.It is well known that the food that edible vitamin content is high, can promote
Into being normally carried out for human metabolism, enhances body to the resistivity of infectious disease, accelerate the healing of wound, and can prevent bad
The generation of blood disease.Containing a kind of substance for being known as heterologous protein in straw mushroom fructification, there is antitumaous effect, the nitrogen leaching contained
Object and purine base can also inhibit the growth of cancer cell out.In addition, summer, which eats straw mushroom, has the function of sunstroke prevention heat-clearing.
Straw mushroom growth hobby hot and humid environment, is suitble to cultivate in tropical and subtropical region, the unexpected fluctuation pole of temperature is not
Conducive to the growth and development of straw mushroom, the quality and yield of straw mushroom is greatly influenced.In addition, the growth of straw mushroom also need it is higher wet
Degree, due to China's North of Yangtze River area, most of the time temperature is lower, dry, and the time for adapting to straw mushroom plantation is extremely short, no
Conducive to extensive cultivation, this greatlys restrict the cultivation range of straw mushroom, therefore city is also much not achieved in the current yield of straw mushroom
The requirement of field, the supply of good protein is not met by the growing demand of consumer especially in straw mushroom.But it closes at present
It is seldom in the research for the protein content for improving straw mushroom.The method for improving protein content existing at present has through addition 30
That there are straw mushroom amino acid contents is relatively low for the existing culture volvaria volvacea cultivation technology of alkanol, vitamin B1, potassium dihydrogen phosphate, mycelia is short
It is small.The disadvantages of density is low, the speed of growth is slow.In addition, it is existing detection straw mushroom in amino acid content method accuracy compared with
It is low, it not can reflect amino acid actual content in straw mushroom.
In consideration of it, the present invention is specifically proposed.
Summary of the invention
The purpose of the present invention is to provide a kind of straw mushroom mediums, and grass can be improved using the straw mushroom medium culture straw mushroom
The amino acid content of mushroom.
The purpose of the present invention is to provide the preparation methods of above-mentioned straw mushroom medium.
Another object of the present invention is to provide a kind of cultural method of straw mushroom, which can be improved the ammonia of straw mushroom
Base acid content.
On the other hand, the present invention also provides a kind of straw mushrooms, have above-mentioned cultural method culture to obtain.
Another object of the present invention is to provide a kind of methods of amino acid content in detection straw mushroom, which can
It more accurately detects the amino acid content in straw mushroom sample, improves the confidence level of testing result.
The invention is realized in this way：
On the one hand, the present invention provides a kind of straw mushroom medium, contain：Potato water extract, glucose, peptone and
Dregs of beans water extract.
Further, in some embodiments of the present invention, by weight, the straw mushroom medium contains：
18-22 parts of glucose, 4-8 parts of peptone and 20-30 parts of dregs of beans water extract, the body of potato water extract and glucose
Product mass ratio is 14-16:1.
Compared to existing ordinary culture medium, culture medium provided by the invention is by being added peptone and dregs of beans water extract
It can make straw mushroom that good hickory chick growth situation be presented, show the feature that mycelia is sturdy, density is big and the speed of growth is fast.
Further, in some embodiments of the present invention, the straw mushroom medium also contains additive, the addition
Agent is selected from one of calcium sulfate, magnesium sulfate, salicylic acid and methyl jasmonate or a variety of combinations.
In further research of the invention, inventors have found that adding calcium sulfate, magnesium sulfate, salicylic acid in the medium
It can also be improved the amino acid content of straw mushroom with any substance in methyl jasmonate.
Further, in some embodiments of the present invention, concentration of the calcium sulfate in straw mushroom medium is 0.5%-
5%.
For example, in some embodiments of the present invention, concentration of the calcium sulfate in straw mushroom medium can be 0.5%,
0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%, 2.5%, 3%, 3.5%, 4%,
4.2%, the range between any one in 4.4%, 4.6%, 4.8% and 5% or both.
Further, in some embodiments of the present invention, concentration of the magnesium sulfate in straw mushroom medium is 1%-
7.5%.
For example, concentration of the magnesium sulfate in straw mushroom medium can be 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%,
2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.1%, 7.2%, 7.3%, 7.4% and
In 7.5% any one or both between range.
Further, in some embodiments of the present invention, concentration of the salicylic acid in straw mushroom medium is
0.00001%-0.01%.
For example, concentration of the salicylic acid in straw mushroom medium can be 0.00001%, 0.0001%, 0.0001%,
In 0.001% and 0.01% any one or both between range.
Further, in some embodiments of the present invention, concentration of the methyl jasmonate in straw mushroom medium is
0.000001%-0.0001%.
For example, concentration of the methyl jasmonate in straw mushroom medium can be 0.000001%, 0.00001% and
In 0.0001% any one or both between range.
Further, in some embodiments of the present invention, the dregs of beans water extract is made as follows：
With the decocting cooked beans dregs of rice, filtering takes filtrate up to the dregs of beans water extract.
Further, in some embodiments of the present invention, when decoction, the mass volume ratio of dregs of beans used and water used
It is 1:22-30.
Further, in some embodiments of the present invention, the dregs of beans water extract is made as follows：25g beans
15-20min is boiled in dregs of rice 500mL boiling, and filtering takes filtrate to get dregs of beans water extract.
Further, in some embodiments of the present invention, the potato water extract is prepared via a method which：It will be native
Beans are mixed with water, are boiled, and filtering takes filtrate, as above-mentioned potato water extract.
Further, in some embodiments of the present invention, the potato water extract is prepared via a method which：It weighs
200g potato is cut into the fritter for being about 3cm, and 500ml water is added, boils 15-20min；Then it is filtered using gauze, takes filter
Liquid, as above-mentioned potato water extract.
On the other hand, the present invention also provides the preparation methods of above-mentioned straw mushroom medium comprising：Potato and water are mixed
It closes, boils, filter, filtrate is taken, as potato water extract；
Glucose, peptone and dregs of beans water extract are mixed with potato water extract, sterilizes, obtains the straw mushroom medium.
It should be noted that can according to circumstances add agar in the case where practical, be prepared into solid medium.
On the other hand, the present invention provides a kind of methods for cultivating straw mushroom comprising following steps：Straw mushroom strain is inoculated with
To straw mushroom medium, the straw mushroom medium contains：Potato water extract, glucose, peptone, agar and dregs of beans water mention
Object.
Further, in some embodiments of the present invention, by weight, the straw mushroom medium contains：
18-22 parts of glucose, 4-8 parts of peptone, 15-20 parts of agar and 20-30 parts of dregs of beans water extract, potato water extract
Volume mass ratio with glucose is 14-16:1.
Compared to existing ordinary culture medium, training cultural method provided by the invention is extracted by the way that peptone and dregs of beans is added
Object can make straw mushroom that good hickory chick growth situation be presented into culture medium, show that mycelia is sturdy, density is big and growth speed
Spend fast feature.
Further, in some embodiments of the present invention, the straw mushroom medium also contains additive, the addition
Agent is selected from one of calcium sulfate, magnesium sulfate, salicylic acid and methyl jasmonate or a variety of combinations.
In further research of the invention, inventors have found that adding calcium sulfate, magnesium sulfate, salicylic acid in the medium
It can also be improved the amino acid content of straw mushroom with any substance in methyl jasmonate.
Further, in some embodiments of the present invention, concentration of the calcium sulfate in straw mushroom medium is 0.5%-
5%.
For example, in some embodiments of the present invention, concentration of the calcium sulfate in straw mushroom medium can be 0.5%,
0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%, 2.5%, 3%, 3.5%, 4%,
4.2%, the range between any one in 4.4%, 4.6%, 4.8% and 5% or both.
Further, in some embodiments of the present invention, concentration of the magnesium sulfate in straw mushroom medium is 1%-
7.5%.
For example, concentration of the magnesium sulfate in straw mushroom medium can be 1%, 1.2%, 1.4%, 1.6%, 1.8%, 2%,
2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.1%, 7.2%, 7.3%, 7.4% and
In 7.5% any one or both between range.
Further, in some embodiments of the present invention, concentration of the salicylic acid in straw mushroom medium is
0.00001%-0.01%.
For example, concentration of the salicylic acid in straw mushroom medium can be 0.00001%, 0.0001%, 0.0001%,
In 0.001% and 0.01% any one or both between range.
Further, in some embodiments of the present invention, concentration of the methyl jasmonate in straw mushroom medium is
0.000001%-0.0001%.
For example, concentration of the methyl jasmonate in straw mushroom medium can be 0.000001%, 0.00001% and
In 0.0001% any one or both between range.
Further, in some embodiments of the present invention, the dregs of beans water extract is made as follows：
With the decocting cooked beans dregs of rice, filtering takes filtrate up to the dregs of beans water extract.
Further, in some embodiments of the present invention, when decoction, the quality volume of dregs of beans used and water used
(g:Ml) than being 1:22-30.
Further, in some embodiments of the present invention, the dregs of beans water extract is made as follows：25g beans
15-20min is boiled in dregs of rice 500mL boiling, and filtering takes filtrate to get dregs of beans water extract.
Further, in some embodiments of the present invention, the potato water extract is prepared via a method which：It will be native
Beans are mixed with water, are boiled, and filtering takes filtrate, as potato water extract.
Further, in some embodiments of the present invention, the potato water extract is prepared via a method which：It weighs
200g potato is cut into the fritter for being about 3cm, and 500ml water is added, boils 15-20min；Then it is filtered using gauze, takes filter
Liquid, as potato water extract.
On the other hand, the present invention also provides a kind of straw mushrooms, are obtained by above-mentioned method culture.
The straw mushroom is full of nutrition, amino acid rich in.
On the other hand, the present invention also provides a kind of methods of amino acid content in detection straw mushroom comprising prepares sample
The sample hydrolysing step and detecting step of product measurement liquid：
The sample hydrolysing step includes：
Step (a)：Hydrochloric acid is added into the hydrolysis pipe equipped with straw mushroom sample, be vortexed concussion, carries out first time hydrolysis, then
Hydrochloric acid is added, be vortexed concussion, carries out second and hydrolyzes, then adds hydrochloric acid again to preset vol；
Volume mass ratio (the mL of the preset vol and straw mushroom sample:It g) is 10-15:1.
It was found by the inventors of the present invention that when measuring amino acid content, by the way that hydrochloric acid is added in sample by stages
When being hydrolyzed, the amino acid in sample can be made more fully to dissolve out, reduce the amino acid content and true amino of detection
Difference between acid content improves the reliability of testing result, improves the reliability of testing result.
Further, in some embodiments of the present invention, when hydrolyzing for the first time, the concentration of hydrochloric acid used is 7.8-
8.2mol/L。
Further, in some embodiments of the present invention, when hydrolyzing for the first time, the volume of hydrochloric acid used is default body
Long-pending 8%-12%.
Further, in some embodiments of the present invention, when second of hydrolysis, the concentration of hydrochloric acid used is 6.8-
7.2mol/L。
Further, in some embodiments of the present invention, when second of hydrolysis, the volume of hydrochloric acid used is default body
Long-pending 8%-12%.
Further, in some embodiments of the present invention, after second hydrolyzes, the concentration of hydrochloric acid added is 5.8-
6.2mol/L。
Further, in some embodiments of the present invention, the protein content of the straw mushroom sample is 10mg-
20mg。
It should be noted that above-mentioned preset vol is 9-11 times of (mL of sample to be tested weight:G), such as 10 times, i.e. 1g sample
Product, corresponding preset vol are 10ml.
It is acid protein hydrolysats matter that the effect of hydrochloric acid is added during analysis of amino acids, passes through the action breaks down albumen of acid
Peptide bond among matter, due to the mycelium that this sample is straw mushroom, which has moisture compared with other dry products or food
Content is higher, the relatively low feature of protein content.In order to increase the extraction efficiency of amino acid, the present invention is special to select ladder
Spend acid hydrolysis method.The advantages of this method be for straw mushroom mycelia cell wall in the component contents such as chitin and cellulose compared with
Moisture content is high in high and sample, therefore first with high concentrated acid cell wall hydrolysis ingredient, protein is discharged, while keeping effective water
The acid concentration for solving peptide bond, improves the extraction efficiency of amino acid.
Further, in some embodiments of the present invention, the sample hydrolysing step further includes following steps：
Step (b)：Interior addition phenol is managed toward hydrolysis, hydrolysis pipe is placed in refrigerant, 3min~5min is freezed, then weighs
Nitrogen is vacuumized-be filled with again, is repeated 3-4 times；
Step (c)：Hydrolysis pipe is placed in 100-120 DEG C of condition to be hydrolyzed, hydrolysis time 20-22h obtains hydrolyzate.
Further, in some embodiments of the present invention, the sample hydrolysing step further includes following steps：
Step (d)：The hydrolyzate is filtered, filtrate is transferred to volumetric flask, scale is settled to water, draws 1.0mL filtrate
It is moved into test tube, is dried under reduced pressure under 40 DEG C~50 DEG C heating environments with test tube concentrating instrument or Parallel evaporator, it is residual after dry
It stays object 1mL~2mL water to dissolve, then is dried under reduced pressure, be evaporated；
Step (e)：In test tube after being added to drying with 1.0mL~2.0mL pH 2.0-2.2 sodium citrate buffer
Dissolution, after oscillation mixes, draw solution is transferred to instrument sample injection bottle after filter membrane, measures liquid as sample.
Further, in some embodiments of the present invention, the detecting step includes：By kilnitamin standard work
Make liquid and sample measurement liquid respectively to pass through calculated by peak area sample with external standard method in same volume injection amino-acid analyzer
Product measure the concentration of amino acid in liquid.
It was found by the inventors of the present invention that when measuring amino acid content, by the way that hydrochloric acid is added in sample by stages
When being hydrolyzed, the protein in sample can be made more fully to dissolve out, avoid the phenomenon that testing result is lower than actual result, have
Conducive to the accuracy for improving testing result.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is by the amino in the Mycelia of Straw Mushroom, Volvariel volvacea turned out using the straw mushroom medium of embodiment 2-5 in experimental example 1
Acid content testing result；
In figure：Ordinate indicates the content (mg/kg) of amino acid, abscissa V1-1, V1-2, V1-3, V1-4, V1-5 difference
The concentration of calcium sulfate is respectively in expression culture medium：0%, 0.5%, 1%, 2%, 5%.
Fig. 2 is by the amino in the Mycelia of Straw Mushroom, Volvariel volvacea turned out using the straw mushroom medium of embodiment 6-9 in experimental example 1
Acid content testing result；
In figure：Ordinate indicates the content (mg/kg) of amino acid, abscissa V2-1, V2-2, V2-3, V2-4, V2-5 difference
The compound concentration of magnesium sulfate is respectively in expression culture medium：0%, 1%, 2.5%, 5%, 7.5%.
Fig. 3 is by the ammonia in the Mycelia of Straw Mushroom, Volvariel volvacea turned out using the straw mushroom medium of embodiment 10-13 in experimental example 1
Base acid content testing result；
In figure：Ordinate indicates the content (mg/kg) of amino acid, abscissa V3-1, V3-2, V3-3, V3-4, V3-5 difference
Indicate that salicylic concentration is respectively in culture medium：0%, 0.00001%, 0.0001%, 0.001%, 0.01%.
Fig. 4 is by the ammonia in the Mycelia of Straw Mushroom, Volvariel volvacea turned out using the straw mushroom medium of embodiment 14-16 in experimental example 1
Base acid content testing result；
In figure：Ordinate indicates the content (mg/kg) of amino acid, and abscissa V4-1, V4-2, V4-3, V4-4 are respectively indicated
The concentration of methyl jasmonate is respectively in culture medium：0%, 0.000001%, 0.00001%, 0.0001%.
Fig. 5 is to be directed to the amino acid that same sample obtains using different analysis of amino acids methods in experimental example 2
Content detection result.
Fig. 6 is the chromatogram of amino standard acid solution；In figure：A is the standard diagram of various amino acid；B is standard diagram
Relevant parameter.
Fig. 7 is that the mycelia during straw mushroom strain is inoculated with respectively on the culture medium of common PDA culture medium and the embodiment of the present invention 1 is raw
Long figure；In figure：A is common PDA culture medium, and B is the culture medium of embodiment 1.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment 1
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
5 parts, 10 parts of agar and 25 parts of dregs of beans water extract.
Wherein, dregs of beans water extract solution is prepared via a method which：
20min is boiled in the 500mL boiling of 25g dregs of beans, and filtering takes filtrate up to dregs of beans water extract solution, about 300mL is spare.
By taking the straw mushroom medium for preparing 1000ml as an example, the preparation method of straw mushroom medium is illustrated, it is as follows：
200g potato is weighed, the fritter for being about 3cm is cut into, 500ml water is added, boils 15-20min；Then gauze is used
It is filtered, takes filtrate, as potato water extract solution.Potato water extract solution 300ml is taken, then takes 20g glucose, 5g albumen
Peptone, 25g dregs of beans water extract solution and 10g agar are added in filtrate, add water polishing to 1000mL, pH naturally, high pressure sterilization,
121 DEG C, 20min.
It is seen using the straw mushroom medium culture medium inoculated straw mushroom strain of the present embodiment using common PDA culture medium as control
The growth situation for examining straw mushroom, is as a result shown in Fig. 7, it can be seen that relatively common PDA culture medium (Fig. 7 A), using the culture of embodiment 1
Base can make straw mushroom that good hickory chick growth situation be presented, and show the feature that mycelia is sturdy, density is big and the speed of growth is fast
(Fig. 7 B).
Embodiment 2
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and mass volume ratio (g:Ml) the calcium sulfate for being 0.5%.
Wherein, by taking the straw mushroom medium for preparing 1000ml as an example, the preparation method of straw mushroom medium is illustrated, such as
Under：
Take potato water extract solution (preparation method is with embodiment 1) 300ml, 20g glucose, 5g peptone, 25g dregs of beans
Water extract, suitable calcium sulphate dihydrate and 10g agar are added in filtrate, add water polishing to 1000mL, and pH is naturally, high pressure is gone out
Bacterium, 121 DEG C, 20min.Wherein, the additive amount of calcium sulphate dihydrate makes it guarantee calcium sulfate in the straw mushroom medium of 1000ml
Final concentration of 0.5%.
Embodiment 3
Straw mushroom medium ingredient provided in this embodiment and preparation method are same as Example 2, the difference is that calcium sulfate
Concentration is 1%.
Embodiment 4
Straw mushroom medium ingredient provided in this embodiment and preparation method are same as Example 2, the difference is that calcium sulfate
Concentration is 2%.
Embodiment 5
Straw mushroom medium ingredient provided in this embodiment and preparation method are same as Example 2, the difference is that calcium sulfate
Concentration is 5%.
Embodiment 6
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
The magnesium sulfate that 5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and mass concentration are 1%.
The preparation method of the straw mushroom medium replaces calcium sulphate dihydrate with embodiment 2, with epsom salt, controls magnesium sulfate
Final concentration of 1% in straw mushroom medium.
Embodiment 7-9
The straw mushroom medium that embodiment 7-9 is provided is substantially the same manner as Example 7, the difference is that the concentration of magnesium sulfate is different,
The magnesium sulfate concentration for the straw mushroom medium that embodiment 7 provides is the magnesium sulfate concentration for the straw mushroom medium that 2.5, embodiment 8 provides
The magnesium sulfate concentration of the straw mushroom medium provided for 5%, embodiment 9 is 7.5%.
Embodiment 10
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and concentration are 0.00001% (mass volume ratio (g:Ml salicylic acid)).
By taking the straw mushroom medium for preparing 1000ml as an example, the preparation method of the straw mushroom medium of the present embodiment is said
It is bright, it is as follows：
200g potato is weighed, suitable water is added, boils 15-20min；Then it is filtered using gauze, takes filtrate；It takes
20g glucose, 5g peptone, 25g dregs of beans water extract, 0.0001g salicylic acid (salicylic acid needs to be first dissolved in organic solvent herein,
Be then added in culture medium) and 10g agar be added in filtrate, add water polishing to 1000mL, pH is naturally, high pressure sterilization, 121
DEG C, 20min.
Wherein, it is as follows to dissolve salicylic method：
1.0000g salicylic acid is accurately weighed, it is 10mL that dehydrated alcohol, which is added, and is dissolved to final volume.1mL is drawn with liquid-transfering gun
Above-mentioned lysate 10 is diluted in the distilled water and alcohol mixed solution (V of heating again：V=5:4) in, sufficiently dissolution is (at this point, bigcatkin willow
Acid concentration is 0.01g/mL).According to salicylic additive amount, with 10 times of gradient dilutions in distilled water.Obtain different salicylic acids
The dilution of content.According to salicylic additive amount, the dilution for choosing appropriate dilutions multiple is added it in culture medium.
For example, taking 10mL concentration is the salicylic acid dilution of 0.0001g/10mL, being then added to final volume control is
In the culture medium of 1000mL, obtain containing 0.00001% salicylic culture medium.
Embodiment 11
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
The salicylic acid that 5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and concentration are 0.0001%.
Embodiment 12
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
The salicylic acid that 5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and concentration are 0.001%.
Embodiment 13
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
The salicylic acid that 5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and concentration are 0.01%.
Embodiment 14
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
The methyl jasmonate that 5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and concentration are 0.000001%.
By taking the straw mushroom medium for preparing 1000ml as an example, the preparation method of the straw mushroom medium of the present embodiment is said
It is bright, it is as follows：
200g potato is weighed, suitable water is added, boils 15-20min；Then it is filtered using gauze, takes filtrate；It takes
20g glucose, 5g peptone, 25g dregs of beans water extract, 0.00001g methyl jasmonate (need first to be dissolved in organic solvent, so
After be added in culture medium) and 10g agar be added in filtrate, add water polishing to 1000mL, pH is naturally, high pressure sterilization, 121 DEG C,
20min。
The method for dissolving methyl jasmonate is as follows：
1mL methyl jasmonate is drawn using liquid-transfering gun to be dissolved in 9mL dehydrated alcohol, is then diluted to distilled water for 10 times
In, obtain the methyl jasmonate dilution that concentration is 0.00001g/10mL.Wherein, extension rate is according to being added to culture medium
In jasmonic acid first dosage determine.
For example, taking 10mL concentration is that the methyl jasmonate dilution of 0.00001g/10mL is all added to final volume control
It is made as in the culture medium of 1000mL, both obtains the culture medium of the methyl jasmonate containing 0.000001% in this way.
Embodiment 15
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
The methyl jasmonate that 5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and concentration are 0.00001%.
Embodiment 16
Straw mushroom medium provided in this embodiment contains 200 parts of potato, 20 parts of glucose, peptone by weight
The methyl jasmonate that 5 parts, 10 parts of agar, 25 parts of dregs of beans water extract and concentration are 0.0001%.
Embodiment 17
Present embodiments provide a kind of method for cultivating straw mushroom comprising：
The strain of straw mushroom V23 is seeded in embodiment 2-16 in any one straw mushroom medium, 37 DEG C of trainings are subsequently placed in
It supports, Mycelia of Straw Mushroom, Volvariel volvacea can be obtained.
Opposite ordinary culture medium can make straw mushroom that good hickory chick be presented and grow state using the culture medium of embodiment 2-16
Gesture shows the feature that mycelia is sturdy, density is big and the speed of growth is fast.
Experimental example 1
The amino acid content in Mycelia of Straw Mushroom, Volvariel volvacea that detection uses the straw mushroom medium of embodiment 2-16 to cultivate.
1 Mycelia of Straw Mushroom, Volvariel volvacea sample collection, method are as follows：
The straw mushroom medium of the embodiment 2-16 for bacterium of having gone out is poured into sterilized and dry culture dish under gnotobasis, to
It is solidified；
Show to stick one layer for the culture medium after solidification to cut and sterilized glassine paper (pays attention to：Glass paper size with
Being slightly less than culture dish is advisable, excessive or too small all unsuitable)；
It is inoculated with straw mushroom V23, every kind of culture medium connects 5 culture dishes to guarantee to be collected into enough mycelia later, trains in 37 DEG C
It supports and is cultivated 3 days in case；Taking-up when it covers with culture dish completely scrapes mycelia, and is collected in 5mL centrifuge tube, deposits in -70
DEG C freezing with spare.
Analysis of amino acids：
The preparation of 2 reagents
Hydrochloric acid solution (6mol/L):It takes 500mL hydrochloric acid to be diluted with water to 1000mL, mixes.
Refrigerant:Commercially available salt is mixed with ice cube by quality 1: 3.
Sodium hydroxide solution (500g/L):50g sodium hydroxide is weighed, is dissolved in 50mL water, it is dilute with water after being cooled to room temperature
It releases to 100mL, mixes.
Sodium citrate buffer [c (Na+)=0.2mol/L]:It weighs 19.6g sodium citrate and the dissolution of 500mL water is added,
16.5mL hydrochloric acid is added, is diluted with water to 1000mL, mixes, is adjusted with 6mol/L hydrochloric acid solution or 500g/L sodium hydroxide solution
PH to 2.2.
The elution buffer solution of different pH and ionic strength:It prepares or buys referring to instrument specification.
3 standard solution are prepared
1) kilnitamin standard reserving solution (1 μm of ol/mL):Single amino acids standard items are accurately weighed respectively (to be accurate to
0.00001g) in same 50mL beaker, is dissolved, is accurately transferred in 250mL volumetric flask with 8.3mL 6mol/L hydrochloric acid solution,
It is diluted with water and is settled to scale, mix, each amino acid standard weighs quality references value and is shown in Table 1.
The weighing quality references value of amino acid standard when table 1 prepares kilnitamin standard reserving solution
2) kilnitamin standard working solution (100nmol/mL):It is accurate to draw kilnitamin standard reserving solution 1.0mL
In 10mL volumetric flask, pH2.2 sodium citrate buffer is added to be settled to scale, mixed, is machine liquid in standard.
4 analyses
1) sample preparation
The Mycelia of Straw Mushroom, Volvariel volvacea of collection step is crushed using tissue pulverizer or grinder, liquor sample is broken into refiner
Homogenate sealing freezen protective, analysis the used time thawed after use.
2) sample weighing
The good sample of uniformity accurately weighs 1g sample (being accurate to 0.0001g) with balance very much, makes protein in sample
Content is within the scope of 10mg~20mg.The sample unknown for protein content can first measure protein content in sample.It will claim
Measured sample is placed in hydrolysis pipe.
3) sample hydrolyzes
(a) enter the 8mol/L hydrochloric acid of 1mL in hydrolysis pipe, vortex 3min is added the 7mol/L hydrochloric acid of 1mL volume, is vortexed
2min is supplemented to about 10mL with 6mol/L hydrochloric acid solution again, and (sample and the mass volume ratio of final volume are 1:10(g/mL)).
It is acid protein hydrolysats matter that the effect of hydrochloric acid is added during analysis of amino acids, passes through the action breaks down albumen of acid
Peptide bond among matter, due to the mycelium that this sample is straw mushroom, which has moisture compared with other dry products or food
Content is higher, the relatively low feature of protein content, and in order to increase the extraction efficiency of amino acid, the present invention is special to select ladder
Spend acid hydrolysis method, the advantages of this method be for straw mushroom mycelia cell wall in the component contents such as chitin and cellulose compared with
Moisture content is high in high and sample, therefore first with high concentrated acid cell wall hydrolysis ingredient, protein is discharged, while keeping effective water
The acid concentration for solving peptide bond, improves the extraction efficiency of amino acid.
(b)~4 drop of the drop of phenol 3 is added into hydrolysis pipe.Hydrolysis pipe is put into refrigerant, 3min~5min is freezed, connects
It onto the exhaust tube of vacuum pump, vacuumizes (close to 0Pa), is then charged with nitrogen and is being filled after repetition vacuumizes-be filled with nitrogen 3 times
Nitrogen state lower sealing tightens screw cap.
(c) the hydrolysis pipe sealed is placed in 110 DEG C ± 1 DEG C of electric heating air blast insulating box or hydrolysis furnace, hydrolyzes 22h
Afterwards, it takes out, is cooled to room temperature.
(d) hydrolysis pipe is opened, hydrolyzate is filtered to 50mL volumetric flask, hydrolysis pipe, washing are repeatedly rinsed with a small amount of water
Liquid moves into same 50mL volumetric flask, is finally settled to scale with water, and oscillation mixes.
Accurate absorption 1.0mL filtrate is moved into 15mL or 25mL test tube, with test tube concentrating instrument or Parallel evaporator 45
Be dried under reduced pressure under DEG C heating environment, it is dry after residue 1mL~2mL water dissolve, then be dried under reduced pressure, be finally evaporated.
(e) it dissolves in test tube after being added to drying with 1.5mL pH2.2 sodium citrate buffer, after oscillation mixes, inhales
Solution is taken, after 0.22 μm of filter membrane, is transferred to instrument sample injection bottle, liquid is measured for sample, for Instrument measuring.
4) it detects
Kilnitamin standard working solution and sample measurement liquid inject amino-acid analyzer respectively with same volume,
(Hitachi L-8900Amino Acid Analyzer (Tokyo, Japan)) is surveyed with external standard method by calculated by peak area sample
Determine the concentration of amino acid in liquid.
Chromatography reference conditions：
Chromatographic column：Sulfonic acid cation resin；
Detection wavelength：570nm and 440nm；
Standard solution：
Amino Acids Mixture Standard Solution,Type H(lot TWK6209Wako Pure
Chemical Industries,Ltd.)；
Standard solution is prepared：0.400mL amino acid mixed standard solution is taken to be settled to 10.00mL with water.
Amino-acid analyzer splitter：
4.6mmI.D.×60mm L packed with Hitachi custom ion exchange resin
(Partical size:3 μm), sodium form (product type #2622)；
Deamination column：4.6mm I.D. × 40mm L (product type #2650L)；
Reaction column：4.6mm I.D.x40mm L, built-in inert silicon sand little particle；
Sample volume：20μL；
Flow velocity：0.4ml/min；Column temperature：57℃；React flow velocity：0.35ml/min；Reaction temperature：135℃.
5) result
It the results are shown in Table 2- table 5 and Fig. 1-Fig. 4, Fig. 6.
Amino acid content detection in the Mycelia of Straw Mushroom, Volvariel volvacea that table 2 is turned out using the straw mushroom medium of embodiment 2-5 is tied
Fruit (unit：mg/kg)
Amino acid content detection in the Mycelia of Straw Mushroom, Volvariel volvacea that table 3 is turned out using the straw mushroom medium of embodiment 6-9 is tied
Fruit (unit：mg/kg)
The amino acid content in Mycelia of Straw Mushroom, Volvariel volvacea that table 4 is turned out using the straw mushroom medium of embodiment 10-13 detects
As a result (unit：mg/kg)
The amino acid content in Mycelia of Straw Mushroom, Volvariel volvacea that table 5 is turned out using the straw mushroom medium of embodiment 14-16 detects
As a result (unit：mg/kg)
It can be seen that addition calcium sulfate, magnesium sulfate, salicylic acid or methyl jasmonate from table 2- table 5 and Fig. 1-Fig. 4 result
The amino acid content in Mycelia of Straw Mushroom, Volvariel volvacea can be improved, the methyl jasmonate of 0.0001% concentration, amino acid are especially added
Total amount reaches 33801mg/kg, i.e., every kg Mycelia of Straw Mushroom, Volvariel volvacea contains 33801mg total amino acid, this knot all high compared with other embodiments
Fruit is that the present inventor is unanticipated.
Experimental example 2
Experimental group：The Mycelia of Straw Mushroom, Volvariel volvacea obtained using embodiment 1 analyzes its amino acid content, the analysis method of experimental group
It is essentially identical with the step 2- step 4 in experimental example 1；
Control group：It is identical as experimental group, the difference is that be handled in (a) step in sample hydrolysing step by following：
Disposably 10mL 6mol/L hydrochloric acid solution is added to be hydrolyzed in hydrolysis pipe.
As a result see Fig. 5, it can be seen that the amino acid content of experimental group is higher than control group, the ammonia being indicated above in experimental example 1
Base acid content analysis method can be such that the protein in sample more fully dissolves out into sample measurement solution, improve testing result
Accuracy.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any to repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of straw mushroom medium, which is characterized in that it contains：Potato water extract, glucose, peptone and dregs of beans water mention
Object.
2. straw mushroom medium according to claim 1, which is characterized in that by weight, the straw mushroom medium contains：
18-22 parts of glucose, 4-8 parts of peptone and 20-30 parts of dregs of beans water extract, the volume matter of potato water extract and glucose
Amount is than being 14-16:1.
3. straw mushroom medium according to claim 2, which is characterized in that the straw mushroom medium also contains additive, institute
It states additive and is selected from one of calcium sulfate, magnesium sulfate, salicylic acid and methyl jasmonate or a variety of combinations.
4. straw mushroom medium according to claim 3, which is characterized in that concentration of the calcium sulfate in straw mushroom medium is
0.5%-5%.
5. straw mushroom medium according to claim 3, which is characterized in that concentration of the magnesium sulfate in straw mushroom medium is
1%-7.5%.
6. straw mushroom medium according to claim 1-5, which is characterized in that the dregs of beans water extract presses such as lower section
Method is made：
With the decocting cooked beans dregs of rice, filtering takes filtrate up to the dregs of beans water extract.
7. straw mushroom medium according to claim 6, which is characterized in that when decoction, the quality of dregs of beans used and water used
Volume ratio is 1:22-30.
8. such as the preparation method of the described in any item straw mushroom mediums of claim 1-7, which is characterized in that it includes：
Potato is mixed with water, is boiled, is filtered, is taken filtrate, obtain potato water extract；
Glucose, peptone and dregs of beans water extract are mixed with potato water extract, sterilizes, obtains the straw mushroom medium.
9. a kind of cultural method of straw mushroom, which is characterized in that it includes：Straw mushroom strain is seeded to any one of claim 1-8
In the straw mushroom medium.
10. a kind of straw mushroom, which is characterized in that it is obtained by cultural method culture as claimed in claim 9.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111418443A (en) * | 2020-05-08 | 2020-07-17 | 青海大学 | Application of salicylic acid or proline in high-temperature cultivation of lentinus edodes |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101933441A (en) * | 2010-09-15 | 2011-01-05 | 上海市农业科学院 | Method for improving yield of straw mushroom |
-
2018
- 2018-08-30 CN CN201811007192.3A patent/CN108901620A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101933441A (en) * | 2010-09-15 | 2011-01-05 | 上海市农业科学院 | Method for improving yield of straw mushroom |
Non-Patent Citations (4)
| Title |
|---|
| 张萍 等: "不同营养条件对草菇液体培养的影响", 《江苏农业科学》 * |
| 李淑娴: "草菇菌丝培养条件优化研究", 《现代食品》 * |
| 王旻: "《生物工程》", 31 August 2015, 中国医药科技出版社 * |
| 肖自添 等: "《草菇优质生产技术》", 30 June 2017, 中国科学技术出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111418443A (en) * | 2020-05-08 | 2020-07-17 | 青海大学 | Application of salicylic acid or proline in high-temperature cultivation of lentinus edodes |
| CN111418443B (en) * | 2020-05-08 | 2021-11-23 | 青海大学 | Application of proline in high-temperature cultivation of lentinus edodes |
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