CN109022288B - Hypsizygus marmoreus solid strain and preparation method thereof - Google Patents
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Abstract
The invention provides a crab-flavored mushroom solid-state strain and a preparation method thereof, belonging to the technical field of edible mushroom production. The preparation method of the solid strain comprises the following steps: 1) preparing a sodium alginate solution containing the Hypsizygus marmoreus liquid strain; 2) adding glutamic acid, chitosan, magnesium stearate and hydroxypropyl cellulose into the sodium alginate solution obtained in the step 1) to perform medium dispersion to obtain mixed solution; 3) and (3) spraying the mixed solution obtained in the step 2) into a calcium chloride exchange solution through a peristaltic pump for embedding treatment, standing and curing, separating out cured particles, and drying to obtain cured strain particles. The preparation method is used for preparing the solid strains of the edible fungi, has simple process and short preparation period, shortens the production period by 18-23 days compared with the traditional solid strains, and is easy for industrial production.
Description
Technical Field
The invention belongs to the technical field related to strains, and particularly relates to a hypsizygus marmoreus solid-state strain and a preparation method thereof.
Background
Hypsizygus marmoreus (Pleurotus eryngii), also known as Hypsizygus Marmoreus, belongs to the sub-phylum Basidiomycotina, class Hymenomycetes, order Agaricales, family Tricholomataceae, genus Hypsizygus. The mushroom not only has beautiful shape, but also has crisp and tender texture, delicious and unique taste and rich nutrition, and has the reputation of 'first crisp in mushrooms'. Commercial planting in China started from the 80 s in the 20 th century, and the commercial planting has become one of the important varieties of edible fungi in factory production in China.
The production process of the edible fungi comprises the preparation of strains, the treatment of a culture medium, the inoculation of cultured fungi, fruiting and the like, wherein the preparation of the strains is particularly critical, the quality of the strains is good and bad, the yield and the quality of the culture are related, and the success or failure of the production is influenced. According to the state of edible fungus strains, the edible fungus strains can be generally divided into liquid strains and solid strains.
The liquid strain has short production period, consistent fungus age, fast spawn running, strong strain activity and regular fruiting after being quantitatively and uniformly sprayed on the material surface. However, once the liquid strains are fermented, the liquid strains must be used immediately, otherwise, the liquid strains are aged for a little time and lose the activity. In addition, the phenomenon of 'double nucleation' sometimes occurs in the process of culturing the liquid strains, and certain risks are brought to the culture.
The solid strains have simple equipment required in the production process, are convenient for production operation and transportation, and are generally accepted by mushroom farmers and edible mushroom cultivation factories. The solid strains include fecal strain, sawdust strain, wood block strain, branch strain, grain strain, granule strain, etc. However, the conventional solid strain preparation process is complex and long in period, so that the strain activity and the mechanical strength are reduced. Therefore, the design of the solid strain with simple preparation process, high strain activity and good mechanical strength has important significance.
Disclosure of Invention
The invention provides a preparation method of a hypsizygus marmoreus solid strain, which solves the technical problems of complex preparation process of the solid strain, long preparation period, insufficient mechanical strength of the obtained solid strain, weak strain activity and the like.
The invention provides a preparation method of a hypsizygus marmoreus solid strain, which comprises the following steps:
1) preparing a sodium alginate solution containing the Hypsizygus marmoreus liquid strain;
2) adding glutamic acid, chitosan, magnesium stearate and hydroxypropyl cellulose into the sodium alginate solution containing the liquid spawn of the Hypsizygus marmoreus obtained in the step 1), and mixing to obtain a mixed solution;
3) and (3) spraying the mixed solution obtained in the step 2) into a calcium chloride exchange solution through a peristaltic pump, standing and solidifying, separating out solidified particles, and drying to obtain solidified strain particles.
Further, in the step 1), the Hypsizygus marmoreus liquid strain is obtained by culturing the Hypsizygus marmoreus liquid strain with a culture medium, wherein the culture medium comprises the following components in parts by weight: corn flour 20-30, yeast extract 1.2-1.5, KH2PO4 0.3-0.5、MgSO4·7H20.5 to 0.7 percent of O and 0.01 to 0.03 percent of clothianidin hydrochloride.
Further, in the step 2), the concentration of sodium alginate in the sodium alginate solution containing the liquid spawn of the Hypsizygus marmoreus is 2-5% (w/v); the content of strain is 1 × 106～1×107cfu/ml。
Further, in the step 2), the addition amount of glutamic acid is 1-3% (w/v), the addition amount of chitosan is 1-3% (w/v), the addition amount of magnesium stearate is 1-3% (w/v), and the addition amount of hydroxypropyl cellulose is 3-5% (w/v).
Further, in the step 3), the concentration of the calcium chloride exchange solution is 3-5% (w/v).
Further, in the step 3), the mixed liquid is sprayed into the calcium chloride exchange liquid by a peristaltic pump at a speed of 18-23 drops/min.
Further, in the step 3), the standing and curing time is 20-40 min.
Further, in step 3), the drying is freeze drying or low-temperature drying.
Further, in the step 3), the freeze drying time is 12-18 h; the low-temperature drying time is 18-24h, and the low-temperature drying temperature is 20-30 ℃.
The invention also provides a solid-state hypsizygus marmoreus strain prepared by the preparation method of the solid-state hypsizygus marmoreus strain.
The preparation method of the solid-state hypsizygus marmoreus strain has the following advantages:
the invention provides a preparation method of a solid-state hypsizygus marmoreus strain, which is characterized in that a liquid strain is solidified to obtain a solid-state strain, and the process is simple to operate, short in preparation period and easy for industrial production. Glutamic acid and chitosan are added into the mixed solution as protective agents, and fillers such as magnesium stearate, hydroxypropyl cellulose and the like are added, so that the activity and the mechanical strength of the strain are further improved.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate an embodiment of the invention and, together with the description, serve to explain the invention and not to limit the invention. In the drawings:
FIG. 1 is a graph showing the germination rate results of solidified seed granules in example 1 of the present disclosure;
FIG. 2 is a graph showing the results of the solid-state strain growth curve measurements in example 1 and a control group;
FIG. 3 is a graph showing the germination rate of solidified seed granules in example 2 of the present disclosure;
FIG. 4 is a graph showing the results of the solid-state strain growth curve measurement in example 2 and the control group;
FIG. 5 is a graph showing the germination rate of solidified seed granules in example 3 of the present disclosure;
FIG. 6 is a graph showing the results of the solid-state strain growth curve measurements in example 3 and a control group;
FIG. 7 is a diagram of solidified bacterial granules obtained in example 1 of the present disclosure;
FIG. 8 is a graph of the germination experiment results of solidified bacterial granules in example 1 of the present disclosure.
Detailed Description
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.
The embodiment of the invention provides a preparation method of a hypsizygus marmoreus solid-state strain, which comprises the following steps:
1) preparing a sodium alginate solution containing the Hypsizygus marmoreus liquid strain;
2) adding glutamic acid, chitosan, magnesium stearate and hydroxypropyl cellulose into the sodium alginate solution containing the liquid spawn of the Hypsizygus marmoreus obtained in the step 1), and mixing to obtain a mixed solution;
3) and (3) spraying the mixed solution obtained in the step 2) into a calcium chloride exchange solution through a peristaltic pump for embedding treatment, standing for solidification, separating, and drying to obtain solidified strain particles.
In the prior art, liquid strains of edible fungi have high viability, but the strains are easy to age and inconvenient to store, and the wide application of the liquid strains is limited, so solid strains are mainly adopted for strain culture in the prior art. However, the traditional solid-state strain preparation process is complex, the period is long, generally about 30-35 days, the difference between the age of the hyphae in the culture medium is large, and when the hyphae at the front end are in a budding state, the hyphae on the surface of the substrate are nearly aged, so that the cultivated mushrooms are not uniform, and the strain activity and the mechanical strength of the strains are reduced.
The embodiment of the invention provides a preparation method of a Hypsizygus marmoreus solid strain. Therefore, the solidified strains are concentrated in density in a certain space by solidifying the liquid strains, so that strain loss is avoided, and the technical problems that the liquid strains are easy to age and the like are effectively solved. Meanwhile, the curing treatment preparation process is simple, the preparation period is short, and the whole process comprises liquid strain preparation and immobilization for about 12 days. The method fully combines the advantages of convenient preparation and high activity of liquid strains and convenient storage and use of solid strains, and is easy to realize factory production and popularization in small workshops of farmers.
In the embodiment of the invention, in the curing process, the mixed solution containing sodium alginate is sprayed into calcium chloride exchange solution through a peristaltic pump for embedding, strains are embedded by utilizing the characteristics that gel formed by the reaction of sodium alginate and calcium salt is insoluble in water and has heat resistance and the like, glutamic acid and chitosan are added into the mixed solution as protective agents, the minimum damage and protection to living cells are realized, and magnesium stearate and hydroxypropyl cellulose are added as filling agents, so that the phenomenon that calcium ions in polyvalent anions and high-concentration electrolytes are easy to fall off is avoided, and the strain activity and the mechanical strength of embedded particles are improved.
It should be noted that W/V (mass to volume) in the examples of the present invention means g/mL, i.e., g/mL in dimension.
The method can also comprise the culture of the liquid spawn of the Hypsizygus marmoreus before the step 1).
Specifically, a suitable hypsizygus marmoreus liquid strain can be obtained by culturing the hypsizygus marmoreus liquid strain by using a culture medium, wherein the culture medium comprises the following components in parts by weight: corn flour 20-30, yeast extract 1.2-1.5, KH2PO4 0.3-0.5、MgSO4·7H20.5 to 0.7 percent of O and 0.01 to 0.03 percent of clothianidin hydrochloride.
The culture of the liquid spawn of the beech mushroom comprises the following steps:
first, the above medium was prepared and sterilized. The method specifically comprises the following steps: preparing the culture medium, sterilizing with 0.1MPa high pressure steam at 120 deg.C for 30min, and culturing in 25 deg.C constant temperature incubator for 24h to obtain sterile culture medium.
Secondly, the strain is inoculated. Inoculating the Hypsizygus marmoreus strain into a sterile culture medium to obtain an inoculation culture medium.
Thirdly, culturing the liquid strain. The method specifically comprises the following steps: placing the inoculated culture medium into a shaker at 25 ℃ for culturing for 4 days to obtain liquid strains.
In the step 1), adding the liquid spawn of the Hypsizygus marmoreus into a sodium alginate solution and mixing to obtain the sodium alginate solution containing the liquid spawn of the Hypsizygus marmoreus.
Wherein, in the sodium alginate solution containing the liquid spawn of the Hypsizygus marmoreus, the concentration of the sodium alginate can be 2-5% (w/v), and specifically can be 2% (w/v), 3% (w/v), 4% (w/v), 5% (w/v) or the like.
The strain content can be 1 × 106～1×107cfu/ml。
In another embodiment, the sodium alginate solution containing the Hypsizygus marmoreus liquid strain can have a sodium alginate concentration of 4% (w/v) and a strain content of 1 × 106～1×107cfu/ml。
In the step 2), glutamic acid, chitosan, magnesium stearate and hydroxypropyl cellulose are added into the sodium alginate solution containing the liquid spawn of the Hypsizygus marmoreus obtained in the step 1) and mixed to obtain a mixed solution.
Further, in step 2), the addition amount of glutamic acid may be 1 to 3% (w/v), the addition amount of chitosan may be 1 to 3% (w/v), the addition amount of magnesium stearate may be 1 to 3% (w/v), and the addition amount of hydroxypropyl cellulose may be 3 to 5% (w/v).
In still another embodiment, in step 2), the addition amount of glutamic acid is 1% (w/v), the addition amount of chitosan is 1% (w/v), the addition amount of magnesium stearate is 1% (w/v), and the addition amount of hydroxypropyl cellulose is 3% (w/v).
In still another embodiment, in step 2), the addition amount of glutamic acid is 3% (w/v), the addition amount of chitosan is 3% (w/v), the addition amount of magnesium stearate is 3% (w/v), and the addition amount of hydroxypropyl cellulose is 5% (w/v).
In the method of the embodiment of the invention, glutamic acid, chitosan, magnesium stearate and hydroxypropyl cellulose are added into the sodium alginate solution for medium dispersion, namely, the sodium alginate solution is physically dispersed. Glutamic acid and chitosan are added as protective agents to realize minimum damage and protection to living cells, and magnesium stearate and hydroxypropyl cellulose are added as filling agents to avoid the phenomenon that polyvalent anions and calcium ions in high-concentration electrolytes are easy to fall off, so that the strain activity and the mechanical strength of embedded particles are improved.
Preferably, in step 2), glutamic acid, chitosan, magnesium stearate and hydroxypropyl cellulose are added and then mixed at a constant temperature to obtain a mixed solution.
Wherein the mixing may be carried out at room temperature; constant temperature mixing at 25-40 deg.C is also possible. Specifically, the constant temperature may be 25 ℃, 30 ℃, 35 ℃, 40 ℃ or the like.
In another embodiment of the present invention, in step 2), glutamic acid, chitosan, magnesium stearate and hydroxypropyl cellulose are added to the sodium alginate solution containing the bacterial strain at a constant temperature of 30 ℃, and mixed to obtain a mixed solution.
In the embodiment of the invention, the specific constant temperature condition is more favorable for spraying the mixed solution into the calcium chloride exchange solution for embedding and curing.
And 3) spraying the mixed solution obtained in the step 2) into a calcium chloride exchange solution through a peristaltic pump, standing and solidifying, separating out solidified particles, and drying to obtain solidified strain particles.
The peristaltic pump can spray the mixed liquid into the calcium chloride exchange liquid at the speed of 18-23 drops/min. During the actual operation, 1 drop is about 0.3-0.5 ml.
The concentration of the calcium chloride exchange solution can be 3-5% (w/v). For example, the calcium chloride exchange solution may be an aqueous solution of calcium chloride at a concentration of 3% (w/v). The concentration of the calcium chloride-exchanged solution may be specifically 4% (w/v), 5% (w/v), or the like.
In the embodiment of the invention, the embedding quality is determined by the speed of the dripping speed (the speed of the peristaltic pump spraying the mixed liquid), the embedding is not easy to be completely embedded at an excessively high speed, and the embedding time is too long due to the excessively low speed, so that the improvement of the activity of the strain is not facilitated.
In the step 3), the standing and curing time can be 20-40min, specifically 20min, 25min, 30min, 35min, 40min and the like.
In the step 3), the solidified particles are separated out and dried to obtain solidified strain particles, namely the required solid strains of the hypsizygus marmoreus.
Suitable drying may be freeze drying or low temperature drying.
Preferably, the low-temperature drying temperature is 20-30 ℃, and the low-temperature drying time is 18-24 h.
Preferably, the freeze-drying time is 12-18 h.
In another embodiment of the present invention, freeze drying may be used to separate the solidified particles and dry them to obtain solidified seed particles. The freeze-drying time may be 12 hours.
Correspondingly, the invention further provides the Hypsizygus marmoreus solid-state strain prepared by any one of the preparation methods of the Hypsizygus marmoreus solid-state strain.
The solid-state hypsizygus marmoreus strain provided by the embodiment of the invention has the advantages of simple preparation process, short period, good mechanical strength and high strain activity.
The present invention will be described in detail with reference to specific examples.
Example 1A production method of a Hypsizygus marmoreus solid strain comprises the following steps:
first, preparation of Hypsizygus marmoreus liquid spawn
Before the preparation of the solid strains, firstly preparing the liquid strains of the hypsizygus marmoreus, which comprises the following steps:
first, a culture medium is prepared and sterilized. The medium composition was as follows: 25 parts of corn flour, 1.3 parts of yeast extract and KH2PO4 0.4、MgSO4·7H2O0.6 and thiamine hydrochloride 0.02. After the preparation of the culture medium, the culture medium is sterilized by high-pressure steam with the pressure of 0.1MPa at 120 ℃ for 30min, and then is cultured in a constant-temperature incubator at 25 ℃ for 1 day to obtain the sterile culture medium.
Secondly, the strain is inoculated. Inoculating Hypsizygus marmoreus strain into sterile culture medium to obtain inoculation culture medium, and inoculating for 5 days.
Thirdly, culturing the liquid strain. Placing the inoculated culture medium into a shaker at 25 ℃ for culturing for 4 days to obtain liquid strains.
Second, preparation of Hypsizygus marmoreus solid strain
1) Preparing sodium alginate solution containing Hypsizygus marmoreus liquid strain
Preparing into sodium alginate solution containing Hypsizygus marmoreus strain, wherein the concentration of the sodium alginate solution is 4% (W/V), and the strain content is 1 × 106～1×107cfu/ml。
2) Adding 1% (w/v) glutamic acid, 1% (w/v) chitosan, 1.5% (w/v) magnesium stearate and 3% (w/v) hydroxypropyl cellulose into the sodium alginate solution containing the hypsizygus marmoreus strain obtained in the step 1), and mixing at constant temperature, wherein the mixing temperature at constant temperature is 25 ℃.
3) And (3) spraying the mixed solution into calcium chloride exchange solution with the concentration of 3-5% (w/v) by a peristaltic pump at the dripping speed of 18 drops/min for embedding, standing and curing for 30min, performing solid-liquid separation by using a screen to take out cured particles, and freeze-drying for 12h to obtain the cured strain particles. The whole course takes about 12 days.
Control groupThe preparation of Hypsizygus marmoreus liquid spawn in example 1 was the same.
The content of the strain obtained in example 1 was 1X 106～1×107cfu/ml, unit volume of 0.007-0.04cm3The diameter of the particles is 2-4mm, and the compression strength of the particles is 3.29kg/cm2The solidified seed particles are shown in FIG. 7.
Example 1 determination of germination rate of solidified seed granules: the solidified strain granules (activated) obtained in example 1 were cultured on PDA plates at 25 ℃ for 4 days, and the measured germination rate was as high as 98% (FIG. 1), and the experimental results of the solidified strain granules germination are shown in FIG. 8.
Growth curve measurements were performed for example 1 and the control group: the immobilized group (the immobilized strain particles (activated) obtained in example 1) and the control group (the liquid strain of Hypsizygus marmoreus) are respectively subjected to growth curve measurement in a liquid culture medium, and comparison shows that the immobilized strain prepared by the process has a good growth state and is similar to the liquid strain proliferation speed (figure 2).
Example 2A production method of a Hypsizygus marmoreus solid strain comprises the following steps:
first, preparation of Hypsizygus marmoreus liquid spawn was performed as in example 1.
Second, preparation of Hypsizygus marmoreus solid strain
1) Preparing sodium alginate solution containing Hypsizygus marmoreus liquid strain
Preparing sodium alginate solution containing Hypsizygus marmoreus strain with concentration of 5% (W/V) and strain content of 1 × 106～1×107cfu/ml。
2) Adding 1% (w/v) glutamic acid, 1.5% (w/v) chitosan, 2% (w/v) magnesium stearate and 4% (w/v) hydroxypropyl cellulose into the sodium alginate solution obtained in the step 1), and mixing at constant temperature to obtain a mixed solution, wherein the temperature for mixing at constant temperature is 30 ℃.
3) And (3) spraying the mixed solution into a calcium chloride exchange solution with the concentration of 3% (w/v) by a peristaltic pump at the dripping speed of 20 drops/min for embedding treatment, standing and curing in the calcium chloride exchange solution for 40min, performing solid-liquid separation by using a screen to take out cured particles, and performing low-temperature drying at the temperature of 25 ℃ for 24h to obtain the cured strain particles. The whole course takes about 12 days.
Example 2 the resulting strain content was 1X 106～1×107cfu/ml, unit volume of 0.007-0.05cm3The diameter of the particles is 2-4mm, and the compression strength of the particles is 3.53kg/cm2。
Example 2 determination of germination rate of solidified seed granules: the solidified seed granules (activated solidified group) obtained in example 2 were cultured on PDA plates at 25 ℃ for 4 days, and the germination rate was determined to be 96% (FIG. 3).
Example 2 and control groups growth curve measurements were performed: the immobilized group (the immobilized strain particles (activated) obtained in example 2) and the control group (the liquid strain of Hypsizygus marmoreus) are respectively subjected to growth curve measurement in a liquid culture medium, and comparison shows that the immobilized strain prepared by the process has a good growth state and is similar to the liquid strain proliferation speed (figure 4).
Example 3A production method of a Hypsizygus marmoreus solid strain comprises the following steps:
first, preparation of Hypsizygus marmoreus liquid spawn was performed as in example 1.
Second, preparation of Hypsizygus marmoreus solid strain
1) Preparing sodium alginate solution containing Hypsizygus marmoreus liquid strain
Preparing sodium alginate solution containing Hypsizygus marmoreus strain with concentration of 5% (W/V) and strain content of 1 × 106～1×107cfu/ml。
2) Adding 1% (w/v) glutamic acid, 1% (w/v) chitosan, 3% (w/v) magnesium stearate and 5% (w/v) hydroxypropyl cellulose into the sodium alginate solution obtained in the step 1), and performing medium dispersion to obtain a mixed solution, wherein the temperature for constant-temperature mixing is 30 ℃.
3) And (3) spraying the mixed solution into 3-5% (w/v) calcium chloride exchange solution by a peristaltic pump at a dripping speed of 18-23 drops/min for embedding, standing and curing in the exchange solution for 30min, performing solid-liquid separation by using a screen to take out cured particles, and drying at the low temperature of 30 ℃ for 18h to obtain the cured strain particles. The whole course takes about 12 days.
The content of the strain obtained in example 3 was 1X 106～1×107cfu/ml, unit volume of 0.007-0.05cm3The diameter of the particles is 2-4mm, and the compression strength of the particles is 3.92kg/cm2。
Example 3 determination of germination rate of solidified seed granules: the solidified seed granules (activated solidified group) obtained in example 3 were cultured on PDA plates at 25 ℃ for 4 days, and the germination rate was determined to be 93% (FIG. 5).
Example 3 and control groups growth curve measurements were performed: the immobilized group (the immobilized strain particles (activated) obtained in example 3) and the control group (the liquid strain of Hypsizygus marmoreus) are respectively subjected to growth curve measurement in a liquid culture medium, and comparison shows that the immobilized strain prepared by the process has a good growth state and is similar to the liquid strain proliferation speed (figure 6).
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions, improvements, etc. within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. A preparation method of a Hypsizygus marmoreus solid strain is characterized by comprising the following steps:
1) preparing a sodium alginate solution containing the Hypsizygus marmoreus liquid strain;
2) adding glutamic acid, chitosan, magnesium stearate and hydroxypropyl cellulose into the sodium alginate solution containing the liquid spawn of the Hypsizygus marmoreus obtained in the step 1), and mixing to obtain a mixed solution;
wherein the addition amount of glutamic acid is 1-3% w/v, the addition amount of chitosan is 1-3% w/v, the addition amount of magnesium stearate is 1-3% w/v, and the addition amount of hydroxypropyl cellulose is 3-5% w/v; in the sodium alginate solution containing the Hypsizygus marmoreus liquid strain, the concentration of the sodium alginate is 2-5% w/v; the content of strain is 1 × 106~1×107cfu/ml；
3) Spraying the mixed solution obtained in the step 2) into a calcium chloride exchange solution through a peristaltic pump, standing and solidifying, separating out solidified particles, and drying to obtain solidified strain particles; wherein the concentration of the calcium chloride exchange solution is 3-5% w/v.
2. The method for preparing the Hypsizygus marmoreus solid-state strain according to claim 1, which is characterized in that: in the step 1), the liquid hypsizygus marmoreus strain is obtained by culturing the hypsizygus marmoreus strain by using the following culture medium, wherein the culture medium comprises the following components in parts by weight: corn flour 20-30, yeast extract 1.2-1.5, KH2PO4 0.3-0.5、MgSO4·7H20.5 to 0.7 percent of O and 0.01 to 0.03 percent of clothianidin hydrochloride.
3. The method for preparing the Hypsizygus marmoreus solid-state strain according to claim 1, which is characterized in that: and 3) spraying the mixed solution into the calcium chloride exchange solution by a peristaltic pump at a speed of 18-23 drops/min.
4. The method for preparing the Hypsizygus marmoreus solid-state strain according to claim 1, which is characterized in that: in the step 3), the standing and curing time is 20-40 min.
5. The method for preparing the Hypsizygus marmoreus solid-state strain according to claim 1, which is characterized in that: in the step 3), the drying is freeze drying or low-temperature drying.
6. The method for preparing the Hypsizygus marmoreus solid-state strain according to claim 5, characterized in that: in the step 3), the freeze drying time is 12-18 h; the low-temperature drying time is 18-24h, and the low-temperature drying temperature is 20-30 ℃.
7. A solid species of hypsizygus marmoreus prepared by the method of any one of claims 1-6.
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