CN105505786A - Culture method for tricholoma matsutake liquid strain - Google Patents
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Abstract
The invention provides a culture method for a tricholoma matsutake liquid strain. The culture method includes the steps that a culture medium for the tricholoma matsutake liquid strain is prepared, the culture medium for the tricholoma matsutake liquid strain is inoculated with a tricholoma matsutake stock seed for sealing culture, and after hyphae of the tricholoma matsutake stock seed germinate, the culture medium for the tricholoma matsutake liquid strain is shaken once every day until the tricholoma matsutake liquid strain is obtained. The culture method for the tricholoma matsutake liquid strain is easy to operate, cultured richoloma matsutake liquid strain hyphae grow fast and are contaminated by other strains little, the hypha output rate is high, and cost is low.
Description
Technical field
The present invention relates to biological technical field, specifically, relate to a kind of cultural method of matsutake liquid strain.
Background technology
Matsutake is the rare wild edible fungus of a kind of preciousness, be of high nutritive value, at present in artificial culture, matsutake bacterial classification still adopts original method of cultivation, cultivating process lacks effective control and management, and nutrition distribution pattern is uneven, needs through multiple steps such as inoculation, quiescent culture and shake-flask culture in matsutake bacterial classification culturing process, but also need manually to pass into sterile air in culture vessel, cause that culturing step is loaded down with trivial details, labour intensity is high.
In order to solve above Problems existing, people are seeking a kind of desirable technical solution always.
Summary of the invention
By Given this, the cultural method of the necessary matsutake liquid strain providing a kind of culturing step simple, with low cost of the present invention, to solve the problem.
The technical solution adopted in the present invention is a kind of cultural method of matsutake liquid strain, specifically comprises the following steps:
A kind of matsutake liquid strain substratum is provided;
Matsutake original seed is inoculated in described matsutake liquid strain substratum and carries out sealing cultivation, after the mycelium germination of described matsutake original seed, shake described matsutake liquid strain substratum every day 1 time, until obtain matsutake liquid strain.
Based on above-mentioned, the step of described matsutake liquid strain substratum is provided to comprise:
Pine branch 200 ~ 300 grams, 200 ~ 400 grams, potato and water 1200 ~ 1800mL are mixed, then successively through boiling, cooling, obtain filtrate after filtration treatment;
Respectively carbon source 10 ~ 150 grams, 10 ~ 200 grams, nitrogenous source are joined in filtrate described in 1000mL, the obtained matsutake liquid strain nutrition filtrate of mixing;
Respectively by 15 ~ 30 grams, husk, KH
2pO
40.2 ~ 2 gram, MgSO
40.1 ~ 2 gram, citric acid 0.1 ~ 3 gram joins in matsutake liquid strain nutrition filtrate described in 150mL and mixes, obtained pH value is the matsutake liquid strain substratum of 5 ~ 7.
Based on above-mentioned, described carbon source is the combination of one or more in lactose, sucrose, Semen Maydis powder, fructose.
Based on above-mentioned, described nitrogenous source is the combination of one or more in yeast extract paste, peptone, extractum carnis, urea.
Preferably, described carbon source is sucrose 10 ~ 80 grams; Described nitrogenous source is bean powder 10 ~ 90 grams.
Based on above-mentioned, after the mycelium germination of described matsutake original seed, timing in every 24 hours shakes described matsutake liquid strain substratum 1 time, and the husk in described matsutake liquid strain substratum is soaked completely.In brief, in the training period, every day, timing shook described morel liquid strain substratum 1 time, such as every morning 9 point, every morning 11 or shake every afternoon 4 described morel liquid strain substratum 1 time, make described husk be immersed in described matsutake liquid strain substratum completely medium.
Based on above-mentioned, the culturing step of described matsutake original seed comprises:
By 15 ~ 25 grams, wheat bran, 20 ~ 30 grams, husk, Semen Maydis powder 8 ~ 12 grams, after being soaked in water respectively, filtering, add water boil respectively, filter and drain, the then obtained compound of mixing; Glucose 5 ~ 10 grams, yeast powder 1 ~ 2 gram, 1 ~ 5 gram, calcium carbonate, 1 ~ 5 gram, phosphate fertilizer is added in described compound; The obtained matsutake pedigree seed culture medium of mixing;
Described matsutake mother is planted to be inoculated in described matsutake pedigree seed culture medium and cultivates, obtain described matsutake original seed.
Based on above-mentioned, the culturing step that described matsutake mother plants comprises:
50 ~ 60 grams, potato, 8 ~ 14 grams, agar, peptone 0.5 ~ 0.8 gram and glucose 5 ~ 7 grams are joined in the water of 1500mL ~ 1800mL and heats, until described agar all boiling, obtain matsutake mother culture media;
The mycelia of matsutake bacterial strain is linked in described matsutake mother culture media and cultivates, obtain described matsutake mother and plant.
In order to better understand the growing state of different times morel bacterial classification, determining liquid incubation time, is within the scope of 0 ~ 240h in the cultivation time, utilizes special sampling thief 24h to sample 1 time, measures the mycelium dry weight of described matsutake bacterial classification.Concrete operation step is: after the per stage cultivation of matsutake bacterial classification terminates, sampling thief is selected to extract liquid appropriate in per stage culturing bottle, take rotating speed as the centrifugation 15min of 4000r/min, then clear liquid is discarded, the described tricholoma matsutake mycelium distilled water obtained is cleaned for several times, be placed in vacuum drying oven, dry at 40 DEG C to constant weight, calculate the described tricholoma matsutake mycelium dry weight of cultivating and obtaining.In experiment, often group carries out 3 parallel tests, and the data obtained gets the mean value of 3 test-results, take incubation time as X-coordinate, and with tricholoma matsutake mycelium dry weight for ordinate zou, draw the growth curve of tricholoma matsutake mycelium, tricholoma matsutake mycelium dry weight calculation formula is as follows:
Y=M/V
In formula; Y represents tricholoma matsutake mycelium dry weight, unit g/L;
M represents the dried quality of mycelium, unit g;
V represents the volume of extracting liquid, unit L.
Hinge structure of the present invention has outstanding substantive distinguishing features and significant progress, specifically, matsutake liquid strain cultural method provided by the present invention, in the process that matsutake liquid strain is cultivated, do not need to shake continuously substratum, only need every day to described matsutake liquid strain substratum shake once, reduce the utilization ratio of culturing process equipment used, thus reduce production cost, and this cultural method is simple, easy handling; Timing shake is adopted every day to ensure that the homogeneity of described matsutake liquid strain nutritive medium.
Further, the growth of matsutake liquid strain mycelia take husk as carrier, the gap between husk is utilized to absorb nutrition, and be attached on the husk on liquid surface and grow, without the need to additionally passing into sterile air in its substratum, just can successfully turn out matsutake liquid strain, decrease the sterile equipment of numerous and diverse operation and many costlinesses, saved cost and shortened cultivation time of bacterial classification.
Further say, the raw material of the liquid strain of matsutake described in the present invention substratum re-uses after liquor, makes the nutriment distribution of described matsutake liquid strain substratum relatively more even, is conducive to matsutake liquid strain mycelia and absorbs; And adopt different culture medium raw materials in each stage of breeding strain, the matsutake bacterial classification mycelia of different cultivation stage fully can be absorbed the nutritive ingredient in substratum, reach best way to cultivate, successfully achieve the artificial culture to matsutake.
Accompanying drawing explanation
Fig. 1 is the mycelium dry weight influence curve figure of the incubation time in embodiment 1 to matsutake liquid strain.
Embodiment
Below by embodiment, technical scheme of the present invention is described in further detail.
The present embodiment provides a kind of cultural method of matsutake liquid strain, specifically comprises the following steps:
Embodiment 1
The 1500mL that pine branch 200 grams, 200 grams, potato added water carries out mixing, boiling and keep 30 points of kinds, then filters, cools obtained filtrate;
In filtrate described in 1000mL, add carbon source, nitrogenous source, get 150mL after fully dissolving and load in culturing bottle, then add 15 grams to described culturing bottle and clean oven dry husk, 0.3 gram of KH
2pO
4, 0.3 gram of MgSO
4with 0.5 gram of citric acid, mix obtained matsutake liquid strain substratum;
The culturing bottle of described matsutake liquid strain substratum will be housed after Autoclave high-temperature sterilization, cooling, be positioned in the aseptic inoculation box of sealing, described matsutake original seed is accessed in described culturing bottle, be placed in 24 DEG C of thermostat containers to cultivate, after the mycelium germination of described matsutake original seed, 9 timings every morning fully shake described matsutake liquid strain substratum 1 time, cultivate certain hour and obtain described matsutake liquid strain.
The cultural method of a kind of matsutake liquid strain provided in the present embodiment, in concrete culturing process, ambient conditions can produce considerable influence to the growth of described matsutake liquid strain; Be further elaborated from the initial pH value of described matsutake liquid strain substratum, carbon source, the growth tendency impact of nitrogenous source aspect on the matsutake liquid strain that the present embodiment provides respectively below.
1. different carbon source is on the impact of tricholoma matsutake mycelium growth tendency
The 1500mL that pine branch 200 grams, 200 grams, potato added water carries out mixing, boiling and keep 30 points of kinds, then filters, cools obtained filtrate;
Respectively to adding the lactose as carbon source of 20g, sucrose, Semen Maydis powder and fructose in filtrate described in 1000mL, getting 150mL after fully dissolving and loading in culturing bottle, then add 15 grams to described culturing bottle and clean and dry husk, 0.3 gram of KH
2pO
4, 0.3 gram of MgSO
4with 0.5 gram of citric acid, this substratum is then utilized to carry out cultivation 5 days to described matsutake original seed, to investigate the impact of different carbon source on described tricholoma matsutake mycelium growth tendency.
Experimental result is as shown in table 1, and as can be seen from the table, the carbon source affecting the liquid mycelial growth trend of described matsutake is followed successively by sucrose, Semen Maydis powder, fructose, lactose from big to small, and therefore preferably sucrose is as the carbon source of matsutake liquid strain substratum; It can also be seen that under only having carbon source not have nitrogenous source situation in substratum, the liquid mycelial growth trend of matsutake is slower simultaneously.
Table 1. different carbon source is on the impact of the liquid mycelial growth trend of matsutake
| Carbon source | Lactose | Sucrose | Semen Maydis powder | Fructose |
| Mycelium dry weight (g/L) | 1.36 | 3.68 | 2.04 | 1.88 |
2. different nitrogen sources is on the impact of mycelial growth trend.
The 1500mL that pine branch 200 grams, 200 grams, potato added water carries out mixing, boiling and keep 30 points of kinds, then filters, cools obtained filtrate;
Respectively to the yeast extract paste as nitrogenous source, peptone, extractum carnis, bean powder, the urea that add 20g in filtrate described in 1000mL, get 150mL after fully dissolving and load in culturing bottle, then add 15 grams to described culturing bottle and clean oven dry husk, 0.3 gram of KH
2pO
4, 0.3 gram of MgSO
4with 0.5 gram of citric acid, this substratum is then utilized to carry out cultivation 5 days to described matsutake original seed, to investigate the impact of different carbon source on described tricholoma matsutake mycelium growth tendency.
Experimental result is as shown in table 2, as can be seen from the table, the nitrogenous source affecting the liquid mycelial growth trend of described matsutake is followed successively by bean powder, extractum carnis, yeast extract paste, urea, peptone from big to small, and therefore preferred bean powder is as the nitrogenous source of matsutake liquid strain substratum.
In table 2. substratum, different nitrogen sources is on the impact of the liquid mycelial growth trend of matsutake
| Nitrogenous source | Yeast extract paste | Peptone | Extractum carnis | Bean powder | Urea |
| Mycelium dry weight (g/L) | 1.30 | 1.05 | 1.32 | 1.59 | 1.14 |
3. initial pH value of medium is on the impact of tricholoma matsutake mycelium growth tendency.
In liquid state fermentation process, because medium component constantly changes, therefore usually only control the initial pH value of substratum.
This experiment adopts substratum based on matsutake liquid strain substratum described in embodiment 1, adopts 20g sucrose and 20g bean powder as the Carbon and nitrogen sources of described substratum respectively, then passes through to regulate the KH in described substratum
2pO
4, MgSO
4, citric acid component proportions carries out the pH value regulating and controlling described substratum.Then utilize respectively pH value be the described matsutake liquid strain substratum of 5,6,7 to described matsutake Primary spawn 5 days, to investigate the impact of initial pH value on tricholoma matsutake mycelium growth tendency.
Test-results is as shown in table 3, as can be seen from the table, along with the increase of initial pH value, described matsutake liquid strain mycelium dry weight occurs first increasing the trend reduced afterwards, as can be seen here, described matsutake liquid strain mycelium suitable growth in the environment of slant acidity, mycelium production is higher, therefore determines that Tricholoma matsutake (lto et lmai) Singer substratum optimal ph is 6.
Table 3. initial pH value of medium is on the impact of the liquid mycelial growth trend of matsutake
| Initial pH value | 5.0 | 6.0 | 7.0 |
| Mycelium dry weight (g/L) | 8.47 | 10.261 | 8.64 |
4. incubation time is on the impact of tricholoma matsutake mycelium growth tendency.
This experiment adopts substratum based on matsutake liquid strain substratum described in embodiment 1, respectively to adding 20g bean powder and 20g sucrose in filtrate described in 1000mL, getting 150mL after abundant dissolving loads in culturing bottle, then adds 15 grams to described culturing bottle and cleans oven dry husk, 0.3 gram of KH
2pO
4, 0.3 gram of MgSO
4the pH value controlling described substratum with 0.5 gram of citric acid is 6, then utilizes this substratum to carry out cultivation certain hour to described matsutake original seed, to investigate the impact of different incubation time on described tricholoma matsutake mycelium growth tendency.
As shown in Figure 1, as can be seen from Fig., in 0 ~ 96h, mycelium dry weight significantly increases along with the prolongation of fermentation time test-results, and when fermentation time is 120h, mycelium dry weight reaches maximum value is 10.51g/L, slowly declines subsequently.Think that, in the fermentation time of 0 ~ 120h, matsutake bacterial classification is in vegetative period, afterwards along with privacies such as nutrition exhaust, matsutake bacterial classification starts self-dissolving and causes production declining.Due to enlarged culturing inoculation is the liquid seeds being in vegetative period, and therefore should be selected in later stage stationary phase of matsutake growth curve, now hypha biomass reaches maximum value, and mycelium does not also occur autolysis.Cultivate matsutake growth curve according to liquid state, the time of finally determining is 120h.
Embodiment 2
The present embodiment provides a kind of cultural method of matsutake liquid strain, and concrete culturing step is roughly the same with embodiment 1, and difference is, the concrete culturing step of matsutake liquid strain comprises:
(1) making of matsutake mother culture media and the cultivation of matsutake mother kind
Successively 50 grams, potato, 8 grams, agar, peptone 0.5 gram, glucose 5 grams are joined in the water of 1500mL and heats, until described agar all boiling, obtain matsutake mother culture media.
Get described matsutake mother culture media 150mL and be placed in culturing bottle and with cotton sealing, being positioned over temperature is sterilizing 2 hours in pressure kettle at 130 DEG C, then naturally cools to room temperature.Described culturing bottle is positioned in the aseptic inoculation box of sealing, in described culturing bottle, accesses the mycelia of matsutake bacterial strain, be placed in 28 DEG C of incubators and cultivate, plant until grow matsutake mother.
(2) making of matsutake pedigree seed culture medium and the cultivation of matsutake original seed
Take 15 grams, wheat bran, 25 grams, husk, Semen Maydis powder 10 grams, glucose 5 grams, yeast powder 1 gram, 2 grams, calcium carbonate and 1.5 grams, phosphate fertilizer respectively.
15 grams, the described wheat bran taken, Semen Maydis powder 10 grams are soaked in water 3 hours respectively, 25 grams, the described husk taken is soaked in water 4 hours, soaked described wheat bran, husk, Semen Maydis powder are placed in pot heating water respectively and boil and keep 2 hours, then pulls out, drain and remove unnecessary moisture.Described wheat bran after draining, husk, Semen Maydis powder are mixed, and add the described glucose 5 grams, yeast powder 1 gram, 2 grams, calcium carbonate and 1.5 grams, the phosphate fertilizer that take wherein, mixes obtained matsutake pedigree seed culture medium.
Be respectively charged in the wide-necked bottle of 100mL by described matsutake pedigree seed culture medium, it is 40g that every bottle of Intake Quantity amounts to dry weight.The polypropylene duplicature of the described wide-necked bottle 10cm × 10cm that will point to install seals a bottle hole, and cutting off a diameter at polypropylene duplicature facing to the position of bottleneck of described wide-necked bottle is the hole of 1.5cm, and seals with two-layer kraft paper, with the fixing sealing of cotton rope.By the sterilizing 15 hours at 120 DEG C of the described wide-necked bottle after sealing, then naturally cooling in sterilisable chamber.Described wide-necked bottle through sterilizing is placed in inoculation tank, then described matsutake mother is planted and be inoculated in described wide-necked bottle according to the inoculum size of every bottle of 2g, and at being placed in 25 DEG C, constant temperature culture obtains matsutake original seed in 12 days.
(3) making of matsutake liquid strain substratum and the cultivation of matsutake liquid strain
Pine branch 200 grams, 200 grams, potato, water 1500mL are carried out mixing, boiling and keep 30 points of kinds, then filters, cool obtained filtrate.
In filtrate described in 1000mL, add 20g sucrose and 20g bean powder successively, get 150mL after fully dissolving and load in culturing bottle, then add 15 grams to described culturing bottle and clean oven dry husk, 0.3 gram of KH
2pO
4, 0.3 gram of MgSO
4with 0.5 gram of citric acid, mix obtained matsutake liquid strain substratum.
The culturing bottle of described matsutake liquid strain substratum will be housed after Autoclave high-temperature sterilization, cooling, be positioned in the aseptic inoculation box of sealing, described matsutake original seed is accessed in described culturing bottle, be placed in 24 DEG C of thermostat containers to cultivate, after the mycelium germination of described matsutake original seed, fully shake described matsutake liquid strain substratum every day 1 time, cultivate and within 5 days, obtain described matsutake liquid strain.
Simultaneous test 1
Simultaneous test 1 provides a kind of cultural method of matsutake liquid strain, concrete culturing step is roughly the same with embodiment 2, difference is, the cultural method of the matsutake liquid strain that the present embodiment provides is quiescent culture, shaking flask process is not carried out to described substratum in culturing process, cultivating 140h test gained matsutake bacterial classification mycelium dry weight is 9.148g/L, and cost is 0.601 yuan/g.
Think, due in culturing process, embodiment 2 has carried out fully rocking to described matsutake liquid strain substratum, dissolved oxygen amount is significantly increased, in addition, due to mycelium depend on husk growth, rock make nutritive ingredient evenly, thus make tricholoma matsutake mycelium growth metabolism vigorous, it is high to produce output.
Simultaneous test 2
Simultaneous test 2 provides a kind of cultural method of matsutake liquid strain, concrete culturing step is roughly the same with embodiment 2, difference is, husk raw material is not comprised in the substratum of the matsutake liquid strain that this simultaneous test provides, and in culturing process, take quiescent culture not carry out shaking flask process to described substratum, cultivating 140h test gained matsutake bacterial classification mycelium dry weight is 8.961g/L, and cost is 0.595 yuan/g.
Simultaneous test 3
Simultaneous test 3 provides a kind of cultural method of matsutake liquid strain, concrete culturing step is roughly the same with embodiment 2, difference is, pine branch and husk raw material is not comprised in the substratum of the matsutake liquid strain that this simultaneous test provides, and in culturing process, take quiescent culture not carry out shaking flask process to described substratum, cultivating 140h test gained matsutake bacterial classification mycelium dry weight is 7.643g/L, and cost is 0.570 yuan/g.
Finally should be noted that: above embodiment is only in order to illustrate that technical scheme of the present invention is not intended to limit; Although with reference to preferred embodiment to invention has been detailed description, those of ordinary skill in the field are to be understood that: still can modify to the specific embodiment of the present invention or carry out equivalent replacement to portion of techniques feature; And not departing from the spirit of technical solution of the present invention, it all should be encompassed in the middle of the technical scheme scope of request of the present invention protection.
Claims (8)
1. a cultural method for matsutake liquid strain, comprises the following steps:
A kind of matsutake liquid strain substratum is provided;
Matsutake original seed is inoculated in described matsutake liquid strain substratum and carries out sealing cultivation, after the mycelium germination of described matsutake original seed, shake described matsutake liquid strain substratum every day 1 time, until obtain matsutake liquid strain.
2. the cultural method of matsutake liquid strain according to claim 1, is characterized in that, provides the step of described matsutake liquid strain substratum to comprise:
Pine branch 200 ~ 300 grams, 200 ~ 400 grams, potato and water 1200 ~ 1800mL are mixed, then successively through boiling, cooling, obtain filtrate after filtration treatment;
Respectively carbon source 10 ~ 150 grams, 10 ~ 200 grams, nitrogenous source are joined in filtrate described in 1000mL, the obtained matsutake liquid strain nutrition filtrate of mixing;
Respectively by 15 ~ 30 grams, husk, KH
2pO
40.2 ~ 2 gram, MgSO
40.1 ~ 2 gram, citric acid 0.1 ~ 3 gram joins in matsutake liquid strain nutrition filtrate described in 150mL and mixes, obtained pH value is the matsutake liquid strain substratum of 5 ~ 7.
3. the cultural method of matsutake liquid strain according to claim 2, is characterized in that, described carbon source is the combination of one or more in lactose, sucrose, Semen Maydis powder, fructose.
4. the cultural method of matsutake liquid strain according to claim 3, is characterized in that, described nitrogenous source is the combination of one or more in yeast extract paste, peptone, extractum carnis, urea.
5. the cultural method of matsutake liquid strain according to claim 4, is characterized in that, described carbon source is sucrose 10 ~ 80 grams; Described nitrogenous source is bean powder 10 ~ 90 grams.
6. the cultural method of the matsutake liquid strain according to any one of Claims 1 to 5, it is characterized in that, after the mycelium germination of described matsutake original seed, timing in every 24 hours shakes described matsutake liquid strain substratum 1 time, and the described husk in described matsutake liquid strain substratum is soaked completely.
7. the cultural method of matsutake liquid strain according to claim 6, is characterized in that, the culturing step of described matsutake original seed comprises:
By 15 ~ 25 grams, wheat bran, 20 ~ 30 grams, husk, Semen Maydis powder 8 ~ 12 grams, after being soaked in water respectively, filtering, add water boil respectively, filter and drain, the then obtained compound of mixing; Glucose 5 ~ 10 grams, yeast powder 1 ~ 2 gram, 1 ~ 5 gram, calcium carbonate, 1 ~ 5 gram, phosphate fertilizer is added in described compound; The obtained matsutake pedigree seed culture medium of mixing;
Described matsutake mother is planted to be inoculated in described matsutake pedigree seed culture medium and cultivates, obtain described matsutake original seed.
8. the cultural method of matsutake liquid strain according to claim 7, is characterized in that, the culturing step that described matsutake mother plants comprises:
50 ~ 60 grams, potato, 8 ~ 14 grams, agar, peptone 0.5 ~ 0.8 gram and glucose 5 ~ 7 grams are joined in the water of 1500mL ~ 1800mL and heats, until described agar all boiling, obtain matsutake mother culture media;
The mycelia of matsutake bacterial strain is linked in described matsutake mother culture media and cultivates, obtain described matsutake mother and plant.
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Cited By (8)
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| CN106212042A (en) * | 2016-07-04 | 2016-12-14 | 李业武 | A kind of pseudo-wild cultivating method of Tricholoma matsutake (lto et lmai) Singer |
| CN106376914A (en) * | 2016-08-25 | 2017-02-08 | 山东天博食品配料有限公司 | Method for preparation of tricholoma matsutake refined powder from tricholoma matsutake submerged fermentation mycelium |
| CN107475137A (en) * | 2017-10-14 | 2017-12-15 | 云南汇林生物科技有限公司 | The breeding method of matsutake strain |
| CN108260470A (en) * | 2018-01-14 | 2018-07-10 | 华中农业大学 | A kind of method for improving matsutake mycorrhizal seedling raising and application |
| CN108967038A (en) * | 2018-08-28 | 2018-12-11 | 铜陵盛牛菌业有限责任公司 | A kind of method of liquid strain cultivation mushroom |
| CN111248026A (en) * | 2020-04-07 | 2020-06-09 | 中国科学院沈阳应用生态研究所 | Quercus matsutake culture medium and application thereof |
| CN112825735A (en) * | 2019-11-25 | 2021-05-25 | 湖南金芙农业科技有限公司 | Tricholoma matsutake liquid strain culture medium and cultivation method of tricholoma matsutake strain |
| CN113862159A (en) * | 2021-10-12 | 2021-12-31 | 天津春发生物科技集团有限公司 | Method for preparing tricholoma matsutake mushroom seasoning by fungus liquid culture method |
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| CN106212042A (en) * | 2016-07-04 | 2016-12-14 | 李业武 | A kind of pseudo-wild cultivating method of Tricholoma matsutake (lto et lmai) Singer |
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| CN108260470A (en) * | 2018-01-14 | 2018-07-10 | 华中农业大学 | A kind of method for improving matsutake mycorrhizal seedling raising and application |
| CN108967038A (en) * | 2018-08-28 | 2018-12-11 | 铜陵盛牛菌业有限责任公司 | A kind of method of liquid strain cultivation mushroom |
| CN112825735A (en) * | 2019-11-25 | 2021-05-25 | 湖南金芙农业科技有限公司 | Tricholoma matsutake liquid strain culture medium and cultivation method of tricholoma matsutake strain |
| CN111248026A (en) * | 2020-04-07 | 2020-06-09 | 中国科学院沈阳应用生态研究所 | Quercus matsutake culture medium and application thereof |
| CN111248026B (en) * | 2020-04-07 | 2022-04-08 | 中国科学院沈阳应用生态研究所 | Quercus matsutake culture medium and application thereof |
| CN113862159A (en) * | 2021-10-12 | 2021-12-31 | 天津春发生物科技集团有限公司 | Method for preparing tricholoma matsutake mushroom seasoning by fungus liquid culture method |
| CN113862159B (en) * | 2021-10-12 | 2024-04-30 | 天津春发生物科技集团有限公司 | Method for preparing tricholoma matsutake seasoning by fungus liquid culture method |
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