CN111264303B - Lyophyllum fumosoroseum cultivar and preparation method thereof - Google Patents
Lyophyllum fumosoroseum cultivar and preparation method thereof Download PDFInfo
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- CN111264303B CN111264303B CN202010275093.4A CN202010275093A CN111264303B CN 111264303 B CN111264303 B CN 111264303B CN 202010275093 A CN202010275093 A CN 202010275093A CN 111264303 B CN111264303 B CN 111264303B
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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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- 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
Abstract
The invention discloses a tobacco-color Lyophyllum karst cultivar and a preparation method thereof. The cultivation species of the fumago-color Lyophyllum karst is obtained by inoculating and culturing a culture material prepared by taking cottonseed hulls, corncobs, sawdust, wheat, gypsum and common calcium as raw materials; the hypha of the culture of the Lyophyllum fumosoroseum has the advantages of fast material consumption, stable properties, low pollution rate and short culture period, and can reduce the phenomenon that the fruiting cannot be normally carried out due to bad strains. The preparation method comprises the steps of pretreatment and inoculation culture. The raw materials used in the invention have simpler composition and low production cost; the culture species of the hon-shimeji mushrooms prepared by the formula and the proportion provided by the invention has the advantages of fast hypha feeding, stable properties, low pollution rate and short culture period, and provides strain guarantee for large-scale culture and sustainable development and utilization of wild hon-shimeji mushrooms in Yunnan.
Description
Technical Field
The invention belongs to the technical field of edible fungus cultivation, and particularly relates to a tobacco-color Lyophyllum karst cultivated species and a preparation method thereof.
Background
Lyophyllum fumosoroseum (Lyophyllum shimeji, Fr.) QuelL . fumosum) Belonging to the family Lyophyllaceae ((Lyophyllaceae), genus Lyophyllum (Lyophyllaceae) ((Lyophyllum)Lyophyllum). The Lyophyllum fumosoroseum is distributed in most counties and cities in Yunnan province, particularly the wild output in regions such as Dali, Chuangxiong, Yuxi, Lincang, Qujing, Pu' er, Jianchuan, Binchuan, Songming and the like is very rich, the wild output usually occurs in 9 months in large quantity, a small quantity of fruiting bodies still occur in the early 11 months in the same year, the fruiting bodies are more in clusters, and the single generation is less, so the fungus is called as psychrophilum, September mushroom, litchis, jungle fowl, northern aeolian fungi and the like by local masses in Yunnan. The wild Lyophyllum fumosoroseum mushroom has the advantages of thick and exquisite meat, delicious taste, more unique flavor after air drying, rich nutrition and dual purposes of medicine and food, is one of important natural resources popular with local people, plays a non-negligible role in improving dietary structure and developing economy, and increases market sales year by year, thus causing the wild resources to be damaged by predation. In recent years, many researchers in China continuously domesticate and cultivate the bacteria, and the bacteria can be cultivated and studied under pure artificial conditionsAnd (5) cultivating the fruiting bodies in rows, and finally cultivating the complete fruiting bodies. However, the artificial cultivation of the Lyophyllum fumosoroseum is still in a small-scale experimental stage, related researches mainly focus on separation identification and biological characteristics, the strain preparation period is long, the character is unstable, the pollution rate is high, the yield of subsequent cultivation is low, and the problems seriously restrict the commercial cultivation of the strain. So far, the artificial cultivation of the Lyophyllum fumosoroseum is hardly seen in the daily market, and the cultivation technology of the Lyophyllum fumosoroseum is still to be perfected.
In the artificial cultivation of the Lyophyllum fumosoroseum, the preparation of excellent strains is a key technical link, the traditional solid strain seed production needs to expand and propagate the produced mother seeds, the original seeds and the cultivated strains step by step, the cultivated strains are often slow in growth or difficult to grow and are easy to infect mixed bacteria, so that a scientific, reasonable, simple-to-operate and easy-to-popularize large-scale preparation method of the Lyophyllum fumosoroseum cultivated strains is necessary to be made, and the phenomena that the mushrooms cannot grow normally and the yield is influenced due to bad strains are reduced.
Disclosure of Invention
The first purpose of the invention is to provide a kind of tobacco color Lyophyllum karst cultivar; the second purpose is to provide the preparation method of the hon-shimeji mushroom cultivar.
The first purpose of the invention is realized by that the cultivar of the fumago Lyophyllum is obtained by inoculating and culturing a culture material prepared by taking cotton seed hulls, corncobs, sawdust, wheat, gypsum and common calcium as raw materials; the hypha of the culture of the Lyophyllum fumosoroseum has the advantages of fast material consumption, stable properties, low pollution rate and short culture period, and can reduce the phenomenon that the fruiting cannot be normally carried out due to bad strains.
The second purpose of the invention is realized by the following steps of pretreatment and inoculation culture, and the method specifically comprises the following steps:
A. pretreatment:
1) weighing fresh, mildew-free, worm-eaten-free and dry raw materials according to a formula ratio for later use;
2) adding gypsum water with the mass concentration of 0.5-1% into corncobs, wheat and cottonseed hulls, soaking for 24-30 h, then washing for 2-3 times by using clear water, and draining to obtain a primary material a of the corncobs, the wheat and the cottonseed hulls;
3) fully and uniformly stirring the primary material a and the sawdust, the gypsum and the common calcium which are proportioned according to the formula to obtain a culture material b;
4) filling the culture material b into a fungus bag, sterilizing and cooling for later use;
B. inoculating and culturing: inoculating the Lyophyllum fumosoroseum strain into a fungus bag, and then culturing at the temperature of 15-25 ℃ to obtain the target Lyophyllum fumosoroseum cultivated species.
The preparation method of the culture medium for the Lyophyllum fumosoroseum cultivar specifically comprises the following steps:
(1) weighing: weighing fresh, mildew-free, worm-eaten-free and dry raw materials according to a formula ratio for later use;
(2) pre-wetting, namely adding gypsum water with the mass concentration of 0.5-1% into the corncobs, the wheat and the cottonseed hulls, soaking for 24-30 h, then washing for 2-3 times by using clear water, and draining to obtain a primary material a of the corncobs, the wheat and the cottonseed hulls;
(3) mixing materials: fully and uniformly stirring the primary material a and sawdust, gypsum and calcium superphosphate which are proportioned according to the formula to obtain a culture material b, wherein the culture material b is required to have no caking, and the water content reaches 60-65%;
(4) bagging: filling the compost b into a polypropylene plastic bag (14 cm multiplied by 28cm multiplied by 0.05 cm), wherein the bagging degree is proper, and the bag opening is sealed by a lantern ring non-cotton cover body;
(5) and (3) sterilization: sterilizing at 121 ℃ (wet heat sterilization) for 2-3 h, transferring the sterilized fungus bags into a cooling chamber sterilized in advance, and timely transferring the sterilized fungus bags into a sterilized (ozone-sterilized) inoculation chamber for later use when the fungus bags are cooled to be less than or equal to 28 ℃.
Wherein the preparation of the cultivar comprises the steps of mother seed propagation, liquid culture and solid culture, and comprises the following steps:
1. culture medium formula
(a) Mother seed propagation culture medium, PDMA culture medium: 200g/L of potato, 15g/L of glucose, 5g/L of maltose and 2g/L, KH of peptone2PO4 1g/L, MgSO40.5g/L and agar 15 g/L.
(b) The liquid culture medium is prepared by mixing PDMA culture medium with agar, and sterilizing at 121 deg.C for 30min (the same as above). Is used for the fermentation culture of the first-stage liquid strains and the fermentation tank strains.
2. Propagation of mother seeds
Taking the test tube mother strain out of a refrigerator at 4 ℃, transferring the test tube mother strain into a plate (with the diameter of 6 cm) filled with a PDMA culture medium for culture, and culturing for 9-11 days at 23-25 ℃ to obtain a plate strain.
3. Liquid strain preparation
The plate strain was inoculated in a liquid medium (potato 200g/L, glucose 15g/L, maltose 5g/L, peptone 2g/L, KH g/L) in a 500mL Erlenmeyer flask by punching 10 round pieces of the strain with a punch having a diameter of 1cm2PO4 1g/L and MgSO40.5g/L), wherein the liquid filling amount per bottle is 200 mL; placing the mixture in a shaking table, and culturing for 5-7 days at the rotating speed of 150-160 r/min and the temperature of 23-25 ℃ to obtain first-stage liquid strains. Transferring the primary liquid strain to a liquid culture medium in a 100L fermentation tank, wherein the liquid filling amount of each tank is 80L, the inoculation amount is 2% -4%, and culturing is carried out for 6-8 days at the rotating speed of 150-160 r/min and the temperature of 23-25 ℃ to obtain the liquid strain.
4. Production of cultivars
Inoculating the liquid strain into a culture medium of a cultivar, wherein the inoculation amount of each bag is 20-25 mL, transferring the cultivar into a culture room for culture, controlling the indoor temperature at 15-25 ℃ and the air relative humidity at 40-60%, and culturing in a dark place to obtain the target cultivar of the Lyophyllum fumigatus.
The invention has the beneficial effects that: the liquid strain is adopted to replace the traditional solid stock, the liquid strain is used as the stock to directly produce the cultivated species, compared with the traditional solid strain production mode, the formula and the manufacturing process of the culture medium are simple, and the strain production cost is reduced; the strain prepared by liquid fermentation has a large number of broken hypha segments and hypha balls, the permeability is good after the strain is inoculated into a culture material of a cultivated species, the hypha segments can quickly permeate into the culture material along with a liquid culture medium, the timely germination and field planting growth after the strain is inoculated are ensured, the strain growth advantage is formed, the phenomenon that the strain is infected with mixed bacteria is reduced, meanwhile, the strain culture period is shortened, and the strain guarantee is provided for the large-scale cultivation and the sustainable development and utilization of the wild Lyophyllum karst in Yunnan local.
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to be limiting in any way, and any modifications or alterations based on the teachings of the present invention are intended to fall within the scope of the present invention.
The cultivated species of the fumago-color Lyophyllum karst is obtained by inoculating and culturing a culture material prepared by taking cottonseed hulls, corncobs, sawdust, wheat, gypsum and common calcium as raw materials; the hypha of the culture of the Lyophyllum fumosoroseum has the advantages of fast material consumption, stable properties, low pollution rate and short culture period, and can reduce the phenomenon that the fruiting cannot be normally carried out due to bad strains.
The culture material comprises, by weight, 40-50 parts of cottonseed hulls, 20-30 parts of corncobs, 10-20 parts of sawdust, 9-17 parts of wheat, 0.5-1.5 parts of gypsum and 0.5-1.5 parts of calcium superphosphate.
The preparation method of the Lyophyllum fumosoroseum cultivar comprises the steps of pretreatment and inoculation culture, and specifically comprises the following steps of:
A. pretreatment:
1) weighing fresh, mildew-free, worm-eaten-free and dry raw materials according to a formula ratio for later use;
2) adding gypsum water with the mass concentration of 0.5-1% into corncobs, wheat and cottonseed hulls, soaking for 24-30 h, then washing for 2-3 times by using clear water, and draining to obtain a primary material a of the corncobs, the wheat and the cottonseed hulls;
3) fully and uniformly stirring the primary material a and the sawdust, the gypsum and the common calcium which are proportioned according to the formula to obtain a culture material b;
4) filling the culture material b into a fungus bag, sterilizing and cooling for later use;
B. inoculating and culturing: inoculating the Lyophyllum fumosoroseum strain into a fungus bag, and then culturing at the temperature of 15-25 ℃ to obtain the target Lyophyllum fumosoroseum cultivated species.
The compost b needs no caking, and the water content is 60-65%.
The fungus bag is a high-temperature-resistant polypropylene plastic bag.
The size of the fungus bag is (12-15) cm multiplied by (24-30) cm multiplied by 0.05 mm.
The sterilization is moist heat sterilization.
The cooling is carried out in a cooling chamber sterilized in advance.
The cooling is to cool the temperature of the strain bag to below 28 ℃.
The culture is carried out in a dark place under the conditions that the temperature is 22-24 ℃ and the relative humidity of air is 40-65%.
The invention is further illustrated by the following specific examples:
example 1
A kind of Lyophyllum fumosoroseum cultivar and its preparation method specifically comprises the following steps:
culture medium formula of cultivar
50% of cottonseed hulls, 20% of corncobs, 19% of sawdust, 9% of wheat, 1% of gypsum and 1% of calcium superphosphate.
(II) preparation method of tobacco-color Lyophyllum karst cultivar
(1) And (3) mother seed propagation: the test tube stock was removed from the refrigerator at 4 ℃ and transferred to a medium containing PDMA (potato 200g/L, glucose 15g/L, maltose 5g/L, peptone 2g/L, KH)2PO4 1g/L, MgSO40.5g/L, agar 15 g/L) in a dish (diameter 6 cm), and culturing at 25 ℃ for 9 days to obtain a dish strain.
(2) Preparing liquid strains: the plate strain was inoculated in a liquid medium (potato 200g/L, glucose 15g/L, maltose 5g/L, peptone 2g/L, KH g/L) in a 500mL Erlenmeyer flask by punching 10 round pieces of the strain with a punch having a diameter of 1cm2PO4 1g/L and MgSO40.5g/L), wherein the liquid filling amount per bottle is 200 mL; placing in a shaking table, and culturing at 25 deg.C and rotation speed of 160r/min for 6 days to obtain first-stage liquid strain. Transferring the first-stage liquid strain to a liquid culture medium (same as above) in a 100L fermentation tank, wherein the liquid filling amount in each tank is 80L, the inoculation amount in each tank is 3%, and the liquid strain is obtained after the culture is carried out for 7 days under the conditions of the rotating speed of 160r/min and the temperature of 25 ℃.
(3) Production of cultivars
A. Weighing: weighing fresh, mildew-free, worm-eaten-free and dry raw materials according to a formula ratio for later use;
B. pre-wetting, namely adding gypsum water with the mass concentration of 0.5% into the corncobs, the wheat and the cottonseed hulls, soaking for 24 hours, then washing for 2 times by using clear water, and draining to obtain primary materials a of the corncobs, the wheat and the cottonseed hulls;
C. mixing materials: fully and uniformly stirring the primary material a and the sawdust, the gypsum and the superphosphate which are proportioned according to the formula to obtain a culture material b, wherein the culture material b is required to have no caking and the water content reaches 65%;
D. bagging: filling the culture material b into a polypropylene plastic bag (14 cm multiplied by 28cm multiplied by 0.05 cm), wherein the tightness of the bag is proper, and the opening of the bag is sealed by a lantern ring non-cotton cover body;
E. and (3) sterilization: sterilizing at 121 deg.C for 2.5h, transferring the sterilized fungus bag into a pre-sterilized cooling chamber, cooling to less than or equal to 28 deg.C, and transferring into a sterilized inoculation chamber;
F. inoculation: inoculating the liquid strains in the fermentation tank into a culture medium of a cultivated species according to an aseptic operation procedure, wherein the inoculation amount of each bag is 25 mL;
G. culturing: after inoculation of the cultivar is completed, the cultivar is moved into a culture room, the indoor temperature is controlled at 23 ℃, the relative air humidity is controlled between 40% and 65%, and the desired cultivar of the Lyophyllum fumosoroseum is obtained by culturing in a dark place.
Example 3 groups of Lyophyllum fumosoroseum cultivars were co-cultured, 100 bags each.
Example 2
To better illustrate the advantages of the present invention, this example was designed as a control, as follows:
(1) in this example, the culturing process and conditions were exactly the same as in example 1 except that 25g of the solid stock was used instead of the liquid seed culture to inoculate the culture medium of the Lyophyllum fumosoroseum cultivar. This example co-cultures 3 groups of Lyophyllum fumosoroseum cultivars, 100 bags per group.
(2) The stock culture medium formula
45% of corncob, 25% of wheat, 18% of sawdust, 1% of gypsum and 1% of ordinary calcium.
(3) The preparation method of the solid stock comprises the following steps:
A. weighing: weighing fresh, mildew-free, worm-eaten-free and dry raw materials according to a formula ratio for later use;
B. pre-wetting, namely adding gypsum water with the mass concentration of 0.5% into the corncobs and the wheat, soaking for 24 hours, then washing for 2 times by using clear water, and draining to obtain primary corncobs and wheat materials c;
C. mixing materials: fully and uniformly stirring the primary material c and the sawdust, the gypsum and the superphosphate which are proportioned according to the formula to obtain a culture material d, wherein the culture material d is required to have no caking and the water content reaches 65%;
D. bottling: the compost is filled into a 650mL tissue culture bottle in a day, the filling tightness is proper, and the bottle is sealed by a matched air-permeable high-temperature-resistant bottle cap;
E. and (3) sterilization: sterilizing at 121 deg.C (wet heat sterilization) for 2.5h, transferring the bottle into a cooling chamber sterilized in advance, cooling to less than or equal to 28 deg.C, and transferring into a sterilized (ozone-sterilized) inoculation chamber in time for use;
F. inoculation: according to the aseptic operation protocol, 10 round bacterium blocks are punched out of the plate strains in the embodiment 1 by a puncher with the diameter of 1cm and are inoculated in a stock culture medium;
G. culturing: after the stock seed inoculation is finished, the stock seed is moved into a culture room, the indoor temperature is controlled to be 23 ℃, the relative air humidity is controlled to be 40-65%, and the solid stock seed is obtained by culturing in a dark place.
(4) The average preparation time of the solid stock is 60 days, which is much longer than the preparation time of the liquid strain in example 1 (13 days); and after the two strains are inoculated in a culture medium of the cultivar, the time required for producing the cultivar by using the liquid strain is less than the time required for producing the cultivar by using the liquid strain (see the table below), thereby shortening the whole preparation period and greatly improving the production efficiency of the Lyophyllum fumosoroseum strain.
| Seed and seed source for cultivation | Germination time/h | Germination Rate/%) | Growth vigor of strains | Average full bag time/day |
| Solid stock seed | 30 | 97 | The hypha is white and dense | 54 |
| Liquid spawn | 24 | 100 | The hypha is white and dense | 45 |
Example 3
A kind of Lyophyllum fumosoroseum cultivar and its preparation method specifically comprises the following steps:
culture medium formula of cultivar
40% of cotton seed hulls, 30% of corncobs, 19% of sawdust, 9% of wheat, 1% of gypsum and 1% of calcium superphosphate.
(II) preparation of Lyophyllum fumosoroseum cultivars is the same as example 1.
Example 4
A kind of Lyophyllum fumosoroseum cultivar and its preparation method specifically comprises the following steps:
culture medium formula of cultivar
43% of cotton seed hulls, 25% of corncobs, 15% of sawdust, 15% of wheat, 1% of gypsum and 1% of calcium superphosphate.
(II) preparation of Lyophyllum fumosoroseum cultivars was performed in the same manner as in example 1.
Claims (1)
1. A preparation method of a hon-shimeji mushroom cultivar is characterized in that the hon-shimeji mushroom cultivar is obtained by inoculating a cultivar culture material consisting of 50% of cottonseed hulls, 20% of corncobs, 19% of sawdust, 9% of wheat, 1% of gypsum and 1% of calcium superphosphate in percentage by weight, and the hon-shimeji mushroom cultivar has the advantages of fast material consumption, stable properties, low pollution rate, short culture period, 24 hours of germination, 100% of germination rate, pure white and compact mycelia, and average bag filling time of 45 days, and can reduce the phenomenon that the mushrooms cannot normally grow due to bad strains; the tobacco-color Lyophyllum karst cultivar is obtained by the following steps:
(1) and (3) mother seed propagation: taking out the test tube mother seeds from a refrigerator at 4 ℃, transferring the test tube mother seeds to a dish filled with a PDMA culture medium consisting of 200g/L of potatoes, 15g/L of glucose, 5g/L of maltose, 2g/L, KH2PO 41 g/L of peptone, 40.5 g/L of MgSO40 and 15g/L of agar for culture, and culturing for 9 days at 25 ℃ to obtain a dish strain;
(2) preparing liquid strains: the plate strain is inoculated to a 500mL Erlenmeyer flask with 200g/L potato, 15g/L glucose, 5g/L maltose and 2g/L, KH peptone2PO4 1g/L、MgSO4Liquid culture medium composed of 0.5g/L, the liquid filling amount per bottle is 200 mL; placing in a shaking table, and culturing at a rotation speed of 160r/min and a temperature of 25 deg.C for 6 days to obtain first-stage liquid strain; transferring the primary liquid strain to a liquid culture medium in a 100L fermentation tank, wherein the liquid filling amount of each tank is 80L, the inoculation amount of each tank is 3%, and the culture time is 7 days under the conditions of the rotating speed of 160r/min and the temperature of 25 ℃ to obtain a liquid strain;
(3) production of cultivars
A. Weighing: weighing fresh, mildew-free, worm-eating-free and dry raw materials according to a cultivation material formula of a cultivated species for later use;
B. pre-wetting: adding gypsum water with the mass concentration of 0.5% into corncobs, wheat and cottonseed hulls, soaking for 24 hours, then washing for 2 times by using clear water, and draining to obtain a primary material a of the corncobs, the wheat and the cottonseed hulls;
C. mixing materials: fully and uniformly stirring the primary material a and the sawdust, the gypsum and the superphosphate which are proportioned according to the formula to obtain a culture material b, wherein the culture material b is required to have no caking and the water content reaches 65%;
D. bagging: filling the culture material b into polypropylene plastic bags of 14cm multiplied by 28cm multiplied by 0.05cm, wherein the bagging degree is proper, and the bag openings are sealed by lantern ring non-cotton covers;
E. and (3) sterilization: sterilizing for 2.5h at 121 deg.C, transferring the sterilized fungus bag into a pre-sterilized cooling chamber, cooling to less than or equal to 28 deg.C, and timely transferring into an inoculation chamber sterilized by ozone for use;
F. inoculation: inoculating the liquid strains in the fermentation tank into a culture medium of a cultivated species according to an aseptic operation procedure, wherein the inoculation amount of each bag is 25 mL;
G. culturing: after inoculation of the cultivar is completed, the cultivar is moved into a culture room, the indoor temperature is controlled at 23 ℃, the relative air humidity is controlled between 40% and 65%, and the desired cultivar of the Lyophyllum fumosoroseum is obtained by culturing in a dark place.
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