CN112538435B - High-yield mushroom SDM-15 and cultivation method thereof - Google Patents
High-yield mushroom SDM-15 and cultivation method thereof Download PDFInfo
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Abstract
The invention relates to a high-yield mushroom SDM-15 and a cultivation method thereof. A high-yield mushroom (Lentinus edodes (Berk.) sing) SDM-15 is preserved in China general microbiological culture Collection center in 2018, 10 months and 28 days, and the address is as follows: the preservation number of the 3 rd storied building of Xilu No.1 of Beijing Korean-Yang district is CGMCC No. 16377. The invention obtains a mushroom (Lentinus edodes (Berk.) sing) SDM-15 by a hybrid and chemical reagent N-methyl-N' -nitro-N-Nitrosoguanidine (NTG) mutagenesis compound mode for the first time, and compared with an original strain Lentinus edodes S84, the strain has larger and thicker mushroom cap, shorter stipe and shortened spawn running time by 4-5 days, and has genetic stability; has better marketability and yield than the prior known bacteria.
Description
Technical Field
The invention relates to a high-yield mushroom SDM-15 and a cultivation method thereof, belonging to the technical field of edible mushroom cultivation.
Background
The mushroom is known as Chinese mushroom, is the edible mushroom variety with the largest output in China, and according to statistics of the Chinese edible mushroom Association, the total output of the mushroom in 2016 years in China reaches 898.3 tons, which accounts for 24.97 percent of the total output of the edible mushrooms in China. The artificial cultivation of the mushrooms originates from Longquan, Qingyuan and Jingning areas in Zhejiang province of China, is a 'flower chopping method', is ancient, has nearly thousand years of history, is emerging as an industry, starts in the early 80 years, starts to develop rapidly in the 90 years, and is at the rising stage at present. The yield is over 150 ten thousand tons, which accounts for more than 13.1 percent of the total yield of the edible fungi.
China is a world big mushroom cultivation country, the cultivation history is long, the cultivation technology is advanced, and particularly the substitute cultivation technology is at the leading level in the world. However, the domestic cultivation of the shiitake mushrooms with the independent intellectual property rights is less, and particularly, the shiitake mushrooms capable of being cultivated in a large area are less. The mushroom substitute cultivation is a main mode for future production and development, and the varieties in the domestic mushroom substitute cultivation are mainly 135 strains and 939 strains at present, and are few.
The mushroom strain is the most important basic production data in the production of mushrooms, and plays a very important role in the yield and quality composition of mushrooms, and particularly in the Chinese mushroom industry developed on the basis of natural ecological environment and traditional agricultural methods, the excellent strain plays a very important role in promoting the technical share occupied by the industry development. The breeding of the mushroom strains in China is synchronously developed with the production of mushrooms, and the breeding of the mushroom strains generally passes through four historical stages, namely a starting stage, an introduction stage, a monospore cross breeding stage and a strain breeding stage by applying a protoplast technology. The strain production in China is still in small workshop production on the whole, and the small strain production determined by decentralized cultivation causes the poor overall strain quality, mixed strains, high growth speed and non-uniformity, and is easy to have adverse accidents of wrong seeds, batch pollution, great yield reduction and the like.
Crossbreeding is a traditional breeding method of organisms, and is a method for gathering the excellent characters of two or more varieties together through mating, and finally obtaining a required new variety after screening and culturing.
N-methyl-N' -nitro-N-Nitrosoguanidine (NTG) is used for mutation breeding, is a well-known chemical super-mutagenic agent with remarkable effect, and has the structural formula:
various strains of different types are obtained by combining gene recombination and induced mutation, so that abundant materials are provided for selection.
Chinese patent document CN106479901A (application No. 201610887201.7) discloses a mushroom strain and its application in mushroom cultivation method. The breeding method is natural breeding, namely, a new strain meeting the production requirement is selected by utilizing beneficial variation generated under natural conditions. (1) The new strain has the advantages of thick and strong hyphae, high growth speed, single fruiting body, large size, roundness, dense fungus flesh, difficult breakage, early fruiting, neat fruiting, few deformed mushrooms, obvious tide number, high biological conversion rate and stable excellent characters. (2) The strain is a wide-temperature substitute for mushroom seeds, and can produce mushrooms in spring and autumn at low temperature and also in summer at high temperature. The application of the shiitake mushroom strain in the shiitake mushroom cultivation method is characterized in that: the method comprises the following specific steps: (1) preparing a culture material, (2) bagging and sterilizing, (3) inoculating, (4) culturing in a fungus bag, (5) and managing fruiting.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a high-yield mushroom SDM-15 and a cultivation method thereof. The high-yield mushroom SDM-15 is obtained by screening in a mode of combining crossbreeding and chemical mutagenesis, has excellent properties, realizes the purpose of high yield and high quality, and is suitable for production in various cultivation modes.
The technical scheme of the invention is as follows:
a high-yield mushroom (Lentinus edodes (Berk.) sing) SDM-15 is preserved in China general microbiological culture Collection center in 2018, 10 months and 28 days, and the address is as follows: the preservation number of the 3 rd storied building of Xilu No.1 of Beijing Korean-Yang district is CGMCC No. 16377.
The preparation method of the mushroom (Lentinus edodes (Berk.) sing) SDM-15 strain comprises the following steps:
(1) performing detoxification treatment on the strains to prepare detoxified thalli;
(2) inoculating the detoxified thallus prepared in the step (1) into a liquid strain culture medium, and performing shake culture for 12-16 days at the temperature of 22-24 ℃ and the speed of 120-160 r/min to prepare liquid seeds;
(3) inoculating the liquid seeds prepared in the step (2) into a solid strain culture medium, culturing at the temperature of 22-24 ℃ until hyphae overgrow the solid strain culture medium, and crushing to obtain the mushroom SDM-15 strain.
Preferably, in step (1), the detoxification step is as follows:
cutting off the front end of the test tube culture medium at 3cm by using inoculation palladium, reserving a 1cm blank area, inoculating mushroom mycelia, culturing at 22-24 ℃ in a dark place until the blank cut area is full of mycelia, taking the mycelia at the cut part to inoculate the mycelia at the blank positions of other cut test tube culture mediums, and repeatedly treating for 3-4 times.
The purpose of the detoxification step is to remove pathogenic bacteria carried by the shiitake mushrooms.
Preferably, in step (2), the liquid strain culture medium comprises the following components:
25-30 g/L glucose, 18-20 g/L bean flour and KH2PO4 2～3g/L，MgSO41.0-2.0 g/L, vitamin B10.05-0.1 g/L, and polyoxypropylene glycerol ether 0.3 ml/L.
Preferably, in the step (2), the solid strain culture medium comprises the following raw materials by weight percent:
78% of wood chips, 20% of bran, 1% of sucrose and 1% of gypsum.
The cultivation method of the shiitake mushroom (Lentinus edodes (Berk.) Sing) SDM-15 comprises the following steps:
(i) bagging the mushroom cultivation material, puncturing, sterilizing and cooling to obtain mushroom sticks;
(ii) inoculating the mushroom SDM-15 strain into the mushroom stick prepared in the step (i), culturing for 20-30 days under the conditions that the temperature is 22-23 ℃ and the humidity is 65% -70%, pricking and oxygenating, then continuously culturing for 40-45 days, pricking and oxygenating again to obtain a mushroom stick with full hyphae;
(iii) and (3) turning the fungus sticks with full hyphae prepared in the step (ii) to color for 30-40 d under the condition that the central temperature of the fungus sticks is not more than 28 ℃ and the scattered light stimulation is carried out for 8h every day at 300-400 lx, removing bags after the fungus sticks are completely turned to be dark brown, fruiting at the temperature of 10-25 ℃, harvesting, recuperating at the temperature of 20-25 ℃ for 10-20 d, injecting water, repeating fruiting, harvesting, recuperating and injecting water again until 4-5 tide of mushrooms are harvested.
According to the invention, in the step (i), the sterilization temperature is 112-113 ℃, and the sterilization time is 4 hours.
According to the invention, in the step (i), the cooling is carried out to the temperature of 18-20 ℃.
Preferably, in the step (iii), water is injected until the weight of the fungus stick is 80-85% of that of the fungus stick before fruiting.
Advantageous effects
The invention obtains a mushroom (Lentinus edodes (Berk.) sing) SDM-15 by a hybrid and chemical reagent N-methyl-N' -nitro-N-Nitrosoguanidine (NTG) mutagenesis compound mode for the first time, and compared with an original strain Lentinus edodes S84, the strain has larger and thicker mushroom cap, shorter stipe and shortened spawn running time by 4-5 days, and has genetic stability; has better marketability and yield than the prior known bacteria.
Drawings
FIG. 1 is a fruiting photograph of Lentinus edodes (Berk.) sing) SDM-15;
FIG. 2 is a fruiting photograph of the original shiitake variety shiitake mushroom (Lentinus edodes (Berk.) Sing) S84;
FIG. 3 is a photograph showing the results of antagonism of SDM-15 of shiitake mushroom (Lentinus edodes (Berk.) Sing) with the original strain;
wherein: the upper part is mushroom S84, and the lower part is mushroom SDM-15.
Detailed Description
The technical solution of the present invention is further described with reference to the following examples, but the scope of the present invention is not limited thereto, and any modifications or equivalent substitutions based on the experimental techniques are within the scope of the present invention.
Bacterial source
Shiitake mushroom (Lentinus edodes (Berk.) sing) SDM-15, deposited in the general microbiological center of the china community collection of microorganisms on 10 and 28 months in 2018, address: the preservation number of the 3 rd storied building of Xilu No.1 of Beijing Korean-Yang district is CGMCC No. 16377.
The PDA culture medium comprises the following components:
200g of potatoes, 50g of sawdust (wrapped by two layers of gauze and boiled together with the potatoes), 20g of glucose, 20g of agar and water to a constant volume of 1L.
Example 1
A strain of mushroom (Lentinus edodes (Berk.) sing) SDM-15 is obtained by screening according to the following method:
(1) selecting naturally mature fruiting body of Lentinus Edodes S84 with good properties, and sterilizingCollecting spores by spore printing method under environment, transferring the collected spores into sterile water after 10h, and diluting 10 times5Dipping spore liquid by using an inoculating loop, coating the spore liquid in a plate culture medium, sealing, and placing the plate culture medium in an environment of 22-24 ℃ for dark culture; when the hyphae grow to 3-6 mm, selecting a small piece of hyphae from the independent bacterial colonies, placing the small piece of hyphae at the central position of a plate culture medium for independent culture at 25 ℃, selecting 8 bacterial colonies with the most vigorous hyphae growth, performing microscopic examination on each bacterial colony when the hyphae grow over an inclined plane, and selecting the hyphae of a monokaryon;
plate culture medium treatment: placing N-methyl-N' -nitro-N-Nitrosoguanidine (NTG) crystals with the diameter of 2mm at the circle center of a culture medium for later use;
(2) selecting 20 mononuclear mycelia for interactive pairing, inoculating the mononuclear mycelia in the center of a culture dish at an inoculation distance of 30mm, and performing constant-temperature dark culture at 25 ℃; detecting the growth distances of two ends of the hyphae respectively in 5, 10 and 15 days, performing microscopic examination on the hyphae with the growth speed of more than or equal to 4.5mm/d, eliminating the rest hyphae, performing microscopic examination on the hyphae with lock-shaped combination, and selecting the hyphae close to the periphery of NTG (nitrilotriacetic acid) for transplantation and propagation culture;
(3) detoxification treatment: cutting off the front end of the test tube culture medium at 3cm by using inoculation palladium and reserving a blank area of 1cm, normally inoculating a mushroom mycelium, culturing the mycelium under the condition of keeping out of the sun at 23 ℃ until the cut-off block culture medium is overgrown over the blank area, inoculating the cut-off block culture medium into other identically treated blank test tubes, and repeatedly treating for 4 times so as to remove pathogenic bacteria carried by mushrooms;
(4) inoculating the mycelium after the selected propagation culture and detoxification into a prepared liquid strain culture medium, performing shake culture for 14 days at the temperature of 23 +/-1 ℃ and at the speed of 150r/min, and inoculating the mycelium into a solid strain culture medium when the growth activity of the mycelium is vigorous;
the liquid strain culture medium comprises the following components:
25g/L glucose, 20g/L bean flour and KH2PO4 2g/L，MgSO42.0g/L, vitamin B10.05g/L and polyoxypropylene glycerol ether 0.3 ml/L;
the solid strain culture medium comprises the following raw materials in percentage by weight:
78% of wood chips, 20% of bran, 1% of sucrose and 1% of gypsum;
(5) and (3) when the bag is full of hypha, mechanically crushing the bag in an aseptic environment, mechanically inoculating the bag into the sterilized mushroom sticks, bagging the mushroom sticks for fungus growth, and performing fruiting culture. And detecting and recording the conditions of fruiting yield, characters, culture period and the like. Repeating the above processes, and finally screening out a strain of Lentinus Edodes (Berk.) Sing) SDM-15 which has larger and thicker mushroom cap, shorter stipe and shortened spawn running time by 4-5 days compared with the original strain Lentinus Edodes S84; the strain is preserved in China general microbiological culture Collection center in 2018, 10 months and 28 days, and the address is as follows: the preservation number of the 3 rd storied building of Xilu No.1 of Beijing Korean-Yang district is CGMCC No. 16377.
Example 2
A preparation method of the Lentinus edodes (Berk.) Sing) SDM-15 seeds comprises the following steps:
(1) cutting off the front end of the test tube culture medium at 3cm by using inoculation palladium and reserving a 1cm blank area, inoculating mushroom mycelia, culturing at 22-24 ℃ in a dark place until the blank cut area is full of mycelia, inoculating the mycelia at the cut position to the blank area of other cut test tube culture mediums, and repeatedly treating for 3 times to obtain detoxified thalli;
(2) inoculating the detoxified thallus prepared in the step (1) into a liquid strain culture medium, and performing shake culture for 14 days at the temperature of 22-24 ℃ and at the speed of 150r/min to prepare liquid seeds;
(3) and (3) inoculating the liquid seeds prepared in the step (2) into a solid strain culture medium, culturing at the temperature of 22-24 ℃ until hyphae overgrow the solid strain culture medium, and crushing to obtain the mushroom SDM-15 seeds.
Preferably, in step (2), the liquid strain culture medium comprises the following components:
27g/L glucose, 18g/L bean flour and KH2PO4 2.5g/L，MgSO41.5 g/L, vitamin B10.075g/L and polyoxypropylene glycerol ether 0.3 ml/L.
Preferably, in the step (2), the solid strain culture medium comprises the following raw materials by weight percent:
78% of wood chips, 20% of bran, 1% of sucrose and 1% of gypsum.
The cultivation method of the shiitake mushroom (Lentinus edodes (Berk.) Sing) SDM-15 comprises the following steps:
(i) bagging the mushroom cultivation material, puncturing, sterilizing for 4 hours at 112-113 ℃, cooling to 18-20 ℃ and preparing mushroom sticks;
(ii) inoculating the mushroom SDM-15 strain into the mushroom stick prepared in the step (i), culturing for 25 days under the conditions that the temperature is 22-23 ℃ and the humidity is 65% -70%, pricking and oxygenating, then continuously culturing for 42 days, pricking and oxygenating again to obtain a mushroom stick with overgrown hyphae;
(iii) and (3) turning the mushroom sticks with full hyphae prepared in the step (ii) to color for 35d under the condition that the central temperature of the mushroom sticks is not more than 28 ℃ and the scattered light of 350lx per day stimulates for 8h, removing bags after the mushroom sticks are completely turned to dark brown, fruiting at the temperature of 18-20 ℃, harvesting, performing rest cultivation at the temperature of 24-25 ℃ for 12d, injecting water until the weight of the mushroom sticks is 82% of that of the mushroom sticks before fruiting, repeating fruiting, harvesting and rest cultivation again, reducing the water injection amount by 15% each time, and harvesting 5-tide mushrooms.
Comparative example
The preparation method as described in example 2, except that the mushroom spawn S84 was used.
Examples of the experiments
Antagonistic experiment of strains
The lentinus edodes strains of the examples and the comparative examples are used as experimental materials, and the used culture medium is PDA culture medium;
the new strains of mushroom, mushroom SDM-15 and mushroom S84, are inoculated and cultured on PDA culture medium in opposite directions, the distance between the inoculation points is 3cm, and the constant temperature culture is carried out for 15 days at 25 ℃.
And (4) analyzing results:
the antagonistic line between SDM-15 and S84 of Lentinus edodes at the boundary of the opposite cultured colonies is evident, and the result is shown in FIG. 2. The screened mushroom (Lentinus edodes (Berk.) sing) SDM-15 has fundamental difference with the original strain, and is a novel mushroom variety with excellent properties, high yield and high quality.
Disease resistance test
Trichoderma viride and Neurospora spp are main diseases in the cultivation process of the lentinus edodes. The pathogenic bacteria Trichoderma viride and Neurospora of Lentinus Edodes are used as indicators for opposite culture, and the disease resistance of Lentinus Edodes SDM-15 is determined by observing the growth of colony and the width of antagonistic zone.
Activating strains: the mushroom strain and the Trichoderma viride strain are respectively placed in a PDA culture medium to be activated and cultured at the temperature of 25 ℃, and the activated and cultured mushroom strain and the Trichoderma viride strain are grown in a culture dish for later use. Neurospora was cultured in PDA medium at 25 ℃ for 2 days and then used as a stock. Before the strain is used, a punch with a diameter of 0.5cm is used to prepare a strain cake.
The method for measuring the trichoderma viride resistance comprises the following steps:
a quantitative culture dish with a diameter of 90mm is used, and 40mL of PDA medium is quantitatively added into each dish by a quantitative peristaltic pump for solidification. Inoculating Lentinus Edodes SDM-15 cake with diameter of 0.5cm 2cm away from the center, culturing at 25 deg.C for 4 days, inoculating Trichoderma viride cake 2cm away from the center, and culturing at 25 deg.C. The width of the antagonistic line, the width of colony growth were measured by observation, and the color and size change of the inhibition zone was continuously observed.
The method for measuring the anti-neurospora capacity comprises the following steps:
a quantitative culture dish with a diameter of 90mm is used, and 40mL of PDA medium is quantitatively added into each dish by a quantitative peristaltic pump for solidification. Inoculating 0.5cm diameter Lentinus Edodes SDM-15 cake at a distance of 1.7cm from the center, inoculating activated Neurospora with inoculating loop at a distance of 1.7cm from the center, and culturing at 25 deg.C for 4 days. The width of the antagonistic line and the width of colony growth were observed and measured.
The results of the disease resistance of the strains are shown in table 1:
TABLE 1
As shown in Table 1, the antagonistic width of mushroom SDM-15 was 4.0mm, which is narrower than the antagonistic width of original strain mushroom S84 by 6.9, in contrast to Trichoderma viride; the antagonistic line width of the mushroom SDM-15 is 4.7mm and is 5.0 less than that of the original strain mushroom S84 in contrast to Trichoderma viride. The disease resistance of the mushroom SDM-15 is better than that of the original strain mushroom S84.
The mushroom SDM-15 has strong capability of resisting trichoderma viride and neurospora. In the industrial cultivation process of the mushroom, bacterial diseases are easy to break out, rotten mushroom diseases appear, and the mushroom SDM-15 strain has strong antibacterial capacity and can reduce the diseases, so that the mushroom cultivation method has a wide application prospect.
Cultivation test
The mushroom (Lentinus edodes (Berk.) sing) SDM-15 and an original strain (Lentinus edodes S84) are adopted to carry out a culture period and culture period optimization experiment in a Lentinus edodes factory cultivation enterprise, the screened strains and the original strains grow 2000 bags of mushrooms respectively, and the whole culture period, the biological conversion rate and the yield are compared:
the cultivation conditions were as follows:
the culture medium comprises the following raw materials in percentage by mass: 78% of wood chips, 20% of bran, 1% of sucrose and 1% of gypsum powder.
Mixing materials: accurately weighing various raw materials prepared on the same day according to the formula proportion. Mixing bran and gypsum, stirring with pre-wetted sawdust, adding water, and stirring to control water content at 55%.
Bagging: after the culture materials are uniformly stirred, the culture materials are sterilized after being filled in the same day. Bagging requires compaction and consistent tightness. And (5) wiping off the culture material at the bag opening part after tying, and tightly tying. And (5) finding broken bags and processing in time.
And (3) sterilization: arranging and placing the bag materials on a specially manufactured bacteria stick rack, pushing the bacteria stick rack into an autoclave, carrying out autoclave sterilization at 116-118 ℃ for 4h, then carrying out heat preservation for 2h, and taking the bags out of the autoclave in time when the temperature of the bags is reduced to 60 ℃.
Inoculation: inoculation was carried out when the bag temperature dropped below 28 ℃. The time is preferably arranged in the evening or in the morning. The operation procedure is as follows: bag wiping and sterilizing → boxing → strain treatment → instrument boxing → sterilizing → inoculation → sealing → box discharging and placing.
And (3) fungus bag culture:
(1) and (4) stacking the mushroom bags, namely arranging the mushroom bags on mushroom fungus rod racks in a Sichuan-shaped order, and paying attention to the placement density of the mushroom fungus racks to facilitate ventilation.
(2) And (3) reasonably pricking and oxygenating: and (4) pricking holes for the first time 15 days after inoculation, wherein 4-6 small holes are pricked around each inoculation hole, and the hole depth is 1 cm. And at 30d for the second time, 8-10 holes are punched around each inoculation hole when the hyphae are connected with each other. And when the fungus bag is completely white for the third time, 20-40 holes are punched, and the depth of the holes is 2-3 cm.
And (3) color conversion period management: and entering a color conversion stage 45-55 days after inoculation. At the moment, the temperature is most suitable to be 20-24 ℃, the relative humidity of the small-environment air is 70% -80%, and the illumination and the fresh air are scattered. Meanwhile, during color changing, the yellow water in the bag is timely removed, so that the fungus cylinder is prevented from being rotten.
Fruiting management, namely a fruiting accelerating technology (1) under the conditions of accelerating fruiting: the temperature is 10-18 ℃ and the temperature difference is more than 10 ℃, the relative humidity of air is 80% -90%, the water content of the fungus bag is 55%, and sufficient oxygen and certain scattered light exist. (2) The mushroom accelerating method comprises the following steps: generally, methods such as vibration, water supplement, temperature difference stimulation and the like are adopted to promote the mushroom bags to grow in batches. (3) Bud breeding: when the mushroom buds grow to 0.5cm, the surrounding film is cut 2/3 in time to expose the mushroom buds. Meanwhile, keeping the temperature in the greenhouse at 8-18 ℃ and the relative humidity of air at 80% -90%, and carrying out bud breeding for 2-3 d. (4) Bud thinning: in order to cultivate high-quality flower mushrooms, reasonably screening over-dense young buds, reserving 7-10 flowers at most in each bag, and enabling the distance between every two flowers to be 3-4 cm.
The flower breeding technology comprises the following steps: (1) flower growing conditions are as follows: the temperature is 12-16 ℃, the temperature difference is more than 10 ℃, the relative humidity of air is controlled to be 50-70%, and scattered light is irradiated. (2) The flower breeding method comprises the following steps: when the young buds are cultivated for 2-3 days in a heat preservation and moisture preservation mode and are 1.5-2 cm, the film around the shed frame is uncovered, the temperature in the shed is reduced, the temperature is controlled, proper ventilation is provided, and cracking of the mushroom cap is promoted.
Harvesting in due time: the mature degree of the mushroom body should be controlled, and the mushrooms are harvested in time according to market requirements. Fresh mushrooms are sold, mushrooms are ripe after growing to 7 minutes, mushroom membranes are not cracked, dry mushrooms are sold, mushrooms are ripe after growing to 8 minutes, mushroom covers are picked when being in bronze gong edges, if in rainy days, flower mushrooms which are not completely ripe are picked in time according to sizes of the flower mushrooms, humidity is prevented from being overlarge, patterns on the surfaces of the mushroom covers are discolored, the whole mushrooms are picked together during picking, small sections of mushroom stems are not remained on mushroom bags, and therefore the mushroom bags are prevented from being mildewed and polluted. After harvest, the results are shown in table 2, with relevant statistical analysis:
TABLE 2
As can be seen from the above table, the fruiting condition, yield and comprehensive evaluation of the mushroom SDM-15 strain all meet the screening requirements, and the character performance is also superior to that of the original strain (mushroom S84). Compared with the original strains, the mushroom SDM-15 shortens the cultivation period. In the industrial cultivation production, the yield of the mushroom SDM-15 is also obviously different, and under the same cultivation fruiting mode, the yield reaches a higher level and is comprehensively evaluated to be excellent.
Claims (9)
1. A high-yield mushroom (Lentinus edodes (Berk.) sing) SDM-15 is preserved in China general microbiological culture Collection center in 2018, 10 months and 28 days, and the address is as follows: the preservation number of the 3 rd storied building of Xilu No.1 of Beijing Korean-Yang district is CGMCC No. 16377.
2. A method for preparing mushroom (Lentinus edodes (Berk.) sing) SDM-15 strain as claimed in claim 1, characterized by the following steps:
(1) performing detoxification treatment on the strains to prepare detoxified thalli;
(2) inoculating the detoxified thallus prepared in the step (1) into a liquid strain culture medium, and performing shake culture for 12-16 days at the temperature of 22-24 ℃ and the speed of 120-160 r/min to prepare liquid seeds;
(3) inoculating the liquid seeds prepared in the step (2) into a solid strain culture medium, culturing at the temperature of 22-24 ℃ until hyphae overgrow the solid strain culture medium, and crushing to obtain the mushroom SDM-15 strain.
3. The method according to claim 2, wherein in the step (1), the detoxification step is as follows:
cutting off the front end of the test tube culture medium at 3cm by using inoculation palladium, reserving a 1cm blank area, inoculating mushroom mycelia, culturing at 22-24 ℃ in a dark place until the blank cut area is full of mycelia, taking the mycelia at the cut part to inoculate the mycelia at the blank positions of other cut test tube culture mediums, and repeatedly treating for 3-4 times.
4. The method according to claim 2, wherein in the step (2), the liquid seed culture medium comprises the following components:
25-30 g/L glucose, 18-20 g/L bean flour and KH2PO4 2～3g/L，MgSO41.0-2.0 g/L, vitamin B10.05-0.1 g/L, and polyoxypropylene glycerol ether 0.3 ml/L.
5. The preparation method according to claim 2, wherein in the step (2), the solid strain culture medium comprises the following raw materials in percentage by weight:
78% of wood chips, 20% of bran, 1% of sucrose and 1% of gypsum.
6. A cultivation method of shiitake mushroom (Lentinus edodes (Berk.) sing) SDM-15 as claimed in claim 1, characterized by the following steps:
(i) bagging the mushroom cultivation material, puncturing, sterilizing and cooling to obtain mushroom sticks;
(ii) inoculating the mushroom SDM-15 strain into the mushroom stick prepared in the step (i), culturing for 20-30 days under the conditions that the temperature is 22-23 ℃ and the humidity is 65% -70%, pricking and oxygenating, then continuously culturing for 40-45 days, pricking and oxygenating again to obtain a mushroom stick with full hyphae;
(iii) and (3) turning the fungus sticks with full hyphae prepared in the step (ii) to color for 30-40 d under the condition that the central temperature of the fungus sticks is not more than 28 ℃ and the scattered light stimulation is carried out for 8h every day at 300-400 lx, removing bags after the fungus sticks are completely turned to be dark brown, fruiting at the temperature of 10-25 ℃, harvesting, recuperating at the temperature of 20-25 ℃ for 10-20 d, injecting water, repeating fruiting, harvesting, recuperating and injecting water again until 4-5 tide of mushrooms are harvested.
7. The cultivation method according to claim 6, wherein in the step (i), the sterilization temperature is 112 to 113 ℃ and the sterilization time is 4 hours.
8. The cultivation method as claimed in claim 6, wherein in the step (i), the cooling is performed to a temperature of 18 to 20 ℃.
9. The cultivation method according to claim 6, wherein in the step (iii), water is injected until the weight of the mushroom stick is 80-85% of that of the mushroom stick before fruiting.
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| CN111876333A (en) * | 2020-06-30 | 2020-11-03 | 四川农业大学 | Lentinus edodes strain XG-3 capable of producing polysaccharide at high yield and application thereof |
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