CN112538432B - Flammulina velutipes strain and cultivation method thereof - Google Patents
Flammulina velutipes strain and cultivation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of edible fungi, and particularly relates to a needle mushroom strain and a cultivation method thereof. The pileus of the strain is golden yellow, the color is bright, the stipe is fine, the average diameter of the stipe is about 0.25cm, the color of the stipe is yellow-white and is not easy to brown, and the base part of the stipe has little villus and is not adhered; after timely harvest, the average lignin content of the stipe is determined to be not higher than 2.50 multiplied by 10 3 A 280nm Per kg, which is significantly lower than the lignin content of the traditional variety (3.05X 10) 3 A 280nm Per kg), the commercial mushroom has good palatability and is suitable for fresh eating; jijin 12 is suitable for an agricultural facility cultivation mode, the yield of fruiting in a single tide and a single bag can reach 288 +/-36.5 g, three tides of mushrooms can be produced through a mushroom intertidal period water supplementing fruiting technology, and the average total biological efficiency is 120-140%.
Description
Technical Field
The invention belongs to the technical field of edible fungi, and particularly relates to a flammulina velutipes strain and a cultivation method thereof.
Background
Needle mushroom（Flammulina filiformis）Belongs to the genus of dried mushrooms, is named as silk cap dried mushrooms in Chinese science and has been used beforeF.velutiper（Fr）SingThe Latin name of the King university is that the Chinese flammulina velutipes is named up to the Kunming plant institute of Chinese academy of sciences 2018 and the Yang wish and the team thereof perform name-identifying on the Chinese flammulina velutipes, and the research proves that the flammulina velutipes in China is different from those in Europe and AmericaF.velutiperIs a new species and should be adoptedF.filiformisThis Latin school name. The needle mushroom is rich in various nutrient substances, and is also called the intelligence-developing mushroom due to the fact that the needle mushroom is rich in lysine. The golden mushroom is divided into yellow and white varieties, wherein yellow is a wild type, and white is a mutant type. Yellow variety ownerThe white variety is mainly cultivated in factories to be cultivated in seasonal agricultural facilities. Yellow varieties are mainly processed, and white varieties are mainly sold fresh.
The main problems of yellow needle mushroom cultivation include two aspects, on one hand, the commercial property is poor, and the defects of easy browning of a traditional yellow needle mushroom variety due to the fact that a mushroom body is exposed to light, more villi in a stipe, serious adhesion of a base part, high toughness (high lignin content) and poor palatability exist, so that the yellow needle mushroom variety is generally required to be processed (for example, pickled) for sale and is rarely sold fresh. However, the loss of nutrients in the flammulina velutipes is serious in the processing processes of pickling and the like, so that a yellow flammulina velutipes variety which is good in marketability and suitable for fresh sale needs to be developed, and the sale way is expanded; on the other hand, the strain is less, the yellow needle mushroom variety for northern agronomic facility cultivation is seriously in short supply, and the yellow needle mushroom variety is always the main cultivated variety Su 6 for many years. In recent years, due to the fact that strains are continuously propagated and lack of scientific and effective strain preservation technology, the traditional strains have the phenomena of strain aging and degeneration, which are reflected in yield reduction, disease resistance reduction and the like; meanwhile, as the cultivation period of mushroom farmers is advanced, the commercial properties of the flammulina velutipes are easy to decline in a low-temperature and low-humidity environment.
Disclosure of Invention
Therefore, the invention aims to overcome the defects of poor commodity, few strains, low yield and the like of the flammulina velutipes in the prior art, and provide a flammulina velutipes strain and a cultivation method thereof.
Therefore, the invention provides the following technical scheme.
The invention provides a golden mushroom strain Jijin 12 with the preservation number of CGMCC No. 19644.
The invention also provides a method for cultivating the flammulina velutipes strain, which comprises the following steps,
(1) and (3) parent seed propagation: inoculating Jijin 12 to a mother strain culture medium, and performing first culture to obtain Jijin 12 mother strains;
(2) stock culture: inoculating the Jijin 12 mother strain on an original strain culture medium, and performing second culture to obtain Jijin 12 original strain;
(3) cultivating cultivars: inoculating the Jijin 12 stock seeds to a cultivated species culture medium, and performing third culture to obtain Jijin 12 cultivated species;
(4) inoculating the Jijin 12 cultivar into a fruiting bag culture medium, and harvesting after hypha culture and fruiting management.
In the step (2), the stock culture medium comprises the following raw materials in a mass ratio of (75-80): (17-21): (0.5-1.5): (0.7-1.3) cottonseed hulls, bran, gypsum and lime, wherein the mass ratio of dry materials to water in the stock culture medium is 1: (1.8-2.3)
In the step (3), the culture medium of the cultivar comprises the following raw materials in mass ratio of (75-80): (17-21): (0.5-1.5): (0.7-1.3) cottonseed hulls, bran, gypsum and lime, wherein the mass ratio of dry materials to water in the culture medium of the cultivated species is 1: (1.8-2.3).
The specific steps of the fruiting management comprise that,
the fruiting bag after mycelium culture enters a bud forcing period, and mushroom buds are promoted to form under the conditions of air relative humidity of 80-90%, temperature of 10-12 ℃, ventilation and low-light induction; mechanical mushroom scratching is carried out on the fruiting bag in the bud forcing period;
when the mushroom buds grow into the mushroom bags, entering a young mushroom period, and growing under the conditions of fresh and circulated air and weak light at the relative air humidity of 85-95% and the temperature of 12-20 ℃ to promote primordial differentiation and branching;
when young mushrooms grow to 2 +/-0.5 cm and enter a mushroom inhibiting period, the young mushrooms grow under the conditions that the relative humidity of air is 80-85%, the temperature is 12-18 ℃, ventilation and scattered light are performed, the thickening of mushroom stems, the thickening of mushroom caps and the robust and neat mushroom bodies are promoted, and the period of the mushroom inhibiting period is 2-3 days;
after the mushroom inhibiting period is finished, spraying water, and growing under the condition that the relative humidity of air is 83-86% to promote the extension of mushroom stems, wherein the period needs to ensure that a mushroom shed is dark and has no light and less ventilation.
In the step (4), the raw materials of the fruiting bag culture medium comprise the following components in percentage by mass (75-80): (13-17): (4-6): (1-3) cottonseed hulls, bran, corn meal and lime, wherein the mass ratio of dry materials to water in the fruiting bag culture medium is 1: (1.8-2.3);
the mass concentration of water in the fruiting bag culture medium is 60-70%;
when the fruiting bag culture medium is inoculated, putting the fruiting bag culture medium into a fungus bag with openings at two ends, sterilizing at 100 ℃ for 12-18h, and then inoculating at two ends of the fungus bag;
requirements of cultivars for inoculation of fruiting bag media: the hypha of the cultivated species is white and strong, and the fungus age is within 15 days after the bottle or the bag is full.
The mycelium culture is carried out at 18-22 deg.C and air relative humidity of 65-70%; in the process of mycelium culture, bags are checked periodically, and if the mycelium is found to be polluted by mixed bacteria, the mycelium is treated immediately to prevent the mycelium from spreading.
The hypha culture and the fruiting management are carried out in a semi-underground fruiting shed;
a plurality of shelf structures are arranged in the fruiting shed;
the ratio of the length, the width and the height of the fruiting shed is 80: 8: (2-3). The number of the layer frame structures arranged in the fruiting shed and the number of the layer frame structures are determined according to the actual cultivation requirements.
In the step (1), the raw materials of the mother culture medium comprise (180-): (19-22): (18-21) peeled potatoes, glucose and agar; the first culture temperature is 20-25 deg.C, air relative humidity is 40-60%, and the time is 5-7 days; the mass ratio of the Jijin 12 to the mother culture medium is (0.8-1.2): 100, respectively; preferably, the raw materials of the mother culture medium comprise 200 parts by weight of peeled potatoes, 20 parts by weight of glucose and 20 parts by weight of agar;
the preparation method of the mother culture medium comprises cutting peeled potato into pieces, boiling in 600ml water for 30min, filtering, collecting filtrate, adding 400ml water, glucose and agar into the filtrate, diluting to 1000ml, heating with slow fire under stirring until the above components are completely dissolved, and sterilizing at 121 deg.C for 30 min.
In the step (2), the temperature of the second culture is 20-25 ℃, the relative humidity of air is 40-60%, and the time is 30-35 days; the mass ratio of the Jijin 12 mother seeds to the stock seed culture medium is (3-5): 100, respectively;
the filling method of the stock culture medium comprises the following steps: subpackaging with a glass bottle and a cottonseed hull bottling machine, wherein the filling amount of the culture medium is 400 +/-50 g, and after culture, one bottle of stock seeds can be used for propagation of 20-25 bottles of cultivated seeds; or the like, or, alternatively,
the strain bags are adopted, the filling amount of the culture medium is 350 +/-50 g after the bag filling machine is used for subpackaging, and after the culture, 15-20 bags of cultivated seeds can be propagated by one bag of stock seeds. The stock culture medium is 1.05kg/cm 2 Sterilizing with high pressure steam for 2.5-3 hr under pressure, and cooling for inoculation.
In the step (3), the temperature of the third culture is 20-25 ℃, the relative humidity of air is 40-60%, and the time is 35-40 days; the mass ratio of the Jijin 12 original seeds to the culture medium of the cultivated species is (3-5): 100, respectively;
the method for filling culture medium of cultivar comprises the following steps: subpackaging with glass bottle and cotton seed hull bottling machine, wherein the filling amount of the culture medium is 400 + -50 g, and after culturing, inoculating 20-30 bags of the cultivated species; or the like, or, alternatively,
the strain bags are adopted, the filling amount of the culture medium is 500 +/-50 g after subpackaging by a bagging machine, and after cultivation, one cultivation strain bag can be inoculated with 30-35 mushroom bags.
In the step (4), when the diameter of the flammulina velutipes pileus is 0.3-0.5cm, the diameter of the stipe is 0.2-0.4cm, and the length of the stipe is 20-25cm, harvesting is carried out.
When the mushroom cultivation bag is collected, the bag opening of the mushroom cultivation bag is pressed by one hand, the mushroom clusters are lightly held by the other hand, the base parts of the fungus stalks are cut in order by a small knife if the fungus stalks are provided with culture mediums, and then the fungus stalks are flatly placed in a plastic basket.
Further, the operation step of tide change management is also included after the harvest;
the number of times of tide change management is 2-3;
after every tide of mushrooms is harvested, water is supplemented, the water supplementing amount is 75-85% of the water loss amount, the operation is tide-changing water supplementing, the fruiting yield of the second tide and the third tide can be improved, and the biological efficiency is improved.
The technical scheme of the invention has the following advantages:
1. the golden mushroom strain Jijin 12 provided by the invention has the preservation number of CGMCC No.19644, the pileus of the strain is golden yellow, the color is bright, the stipe is fine, the average diameter is about 0.25cm, the stipe is yellow-white and is not easy to brown, and the base part of the stipe has less fluff and is not adhered; the average lignin content of the fruiting body stipe in harvesting period is not higher than 2.50 × 10 3 A 280nm Per kg, which is significantly lower than the lignin content of the traditional variety (3.05X 10) 3 A 280nm Per kg), the commercial mushroom has good palatability and is suitable for fresh eating; the Jijin 12 is suitable for an agricultural facility cultivation mode, the yield of single-tide single-bag fruiting can reach 288 +/-36.5 g, three-tide mushrooms can be produced by a mushroom inter-tidal period water supplementing fruiting technology, and the average total biological efficiency is 120-140%.
2. The cultivation method of the flammulina velutipes strain provided by the invention is characterized in that the golden 12 of the flammulina velutipes strain is cultivated, and the average lignin content of the carpopodium of the fruiting body in the harvesting stage is not higher than 2.50 multiplied by 10 by controlling the temperature, the humidity and the culture medium of each link 3 A 280nm The lignin content of the commercial mushroom is obviously lower than that of the conventional variety, the palatability of the commercial mushroom is suitable for fresh eating, and the single-moisture single-bag fruiting yield can reach 288 +/-36.5 g.
Cottonseed hulls, bran, corn flour, gypsum and lime in a specific ratio are used as a stock culture medium, a cultivated species culture medium and a fruiting culture medium, so that the requirements of hypha on vegetative growth can be met, sufficient energy is provided for fruiting, and the yield of needle mushrooms is increased; the culture medium is also beneficial to the golden mushroom to produce three-tide mushrooms, and the tide conversion period is short; the culture medium is also helpful for increasing the water content of needle mushroom, so that the needle mushroom is more brittle and tender.
In the cultivation method, two ends of the mushroom bag are inoculated, which is beneficial to rapid spawn running; the method is beneficial to reducing the mixed bacteria pollution by limiting the parameters and the operation steps of hypha culture. By limiting the operation steps and parameters of fruiting management, the growth environments of the needle mushrooms in different growth periods are controlled, the growth of the stems and the caps of the needle mushrooms is facilitated, the biological efficiency is improved, and the lignin content is reduced. By limiting the harvesting period of the flammulina velutipes, the yield and the commodity of the flammulina velutipes can be improved, the yield can be reduced when the flammulina velutipes are harvested too early, and the commodity of the flammulina velutipes can be influenced when the flammulina velutipes are harvested too late. By adopting the moisture transferring and replenishing technology, the fruiting yield of the second tide and the third tide can be improved, and the total biological efficiency reaches 120-140 percent.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic view of a needle mushroom obtained in example 1 of the present invention.
Detailed Description
The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.
Example 1
A flammulina velutipes strain Jijin 12 is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms, and the address is as follows: the microbial research institute of China academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, with the preservation number of CGMCC No.19644 and the preservation date of 2020, 5 months and 26 days.Classification naming of foil 12: the dried mushrooms are covered with the silk cap, Flammulinafiliformis。
the breeding method of the flammulina velutipes strain comprises the following steps,
collecting yellow mycelium of Flammulina velutipes (Fr.) Sing strain HUANG1Inoculating the cells into a triangular flask containing liquid culture medium, static culturing at 25 deg.C for 8 days, shaking the flask by hand once every 24h, collecting mycelium in liquid culture medium under aseptic condition, adding muramidase enzyme solution (purchased from Guangdong province institute of microorganisms) with mass concentration of 2%, performing enzymolysis at 30 deg.C for 4 hr, filtering to remove broken hypha, washing with 0.6M sucrose solution for 3 times to obtain purified protoplast suspension, diluting with 0.6M sucrose solution to concentration of 300/ml, sucking 0.2ml, spreading on regeneration culture medium, culturing at 25 deg.C for 8 days, selecting single colony, storing on test tube slant, culturing at 25 deg.C for 4 days, collecting mycelium, flaking, observing whether the locked combination exists under a microscope, if the locked combination does not exist, obtaining a protoplast monokaryon, storing the protoplast monokaryon, and taking the protoplast monokaryon as a monokaryon parent, wherein the protoplast monokaryon is marked as HUANG 1-Y-x; wherein the liquid culture medium is prepared from 1L distilled water containing corn flour 3.00% (leaching solution), rhizoma Solani Tuber osi 15.00% (leaching solution), glucose 0.50%, and KH 2 PO 4 0.02%、K 2 HPO 4 0.08%、MgSO 4 0.05 percent; the regeneration medium comprises the following raw materials in parts by weight: adding 200g of potato decoction leachate, 4g of peptone, 23g of agar and 205.2g of sucrose into 1L of distilled water, and keeping the pH value natural.
Collecting fruiting body of wild Flammulina velutipes strain JHH, and performing tissue separation to obtain pure mycelium as binuclear parent.
Inoculating a hypha block of a protoplast mononuclear strain HUANG1-Y-x on one side of a PDA culture medium flat plate, culturing for 4 days at 25 ℃, inoculating an activated hypha block of a binuclear strain JHH with the diameter of 0.5 +/-0.05 cm on the other side of the flat plate when a bacterial colony grows to the diameter of 2 +/-0.1 cm, inoculating the hypha block at the position 1cm away from the edge of the mononuclear hypha, culturing the two hypha in opposite directions until the two hypha are contacted, taking out the hypha block with the diameter of 0.3-0.5cm from the position far away from the binuclear hypha on one side of the mononuclear strain, inoculating the hypha block on a new PDA slant culture medium, culturing in dark light at 25 ℃, and obtaining the strain FHJ-x when the binuclear hypha grows to 1-2 cm.
And (4) performing hypha culture on the binuclear strain FHJ-x, selecting the hybrids with dense hypha and high growth speed, and performing a fruiting test. Uniformly mixing 78g of cottonseed hulls, 15g of bran, 5g of corn meal and 2g of lime, adding 186g of water, and uniformly stirring to obtain a mushroom culture medium; using 17 × 33 × 0.05cm high-density polypropylene plastic as a fruiting bag, filling a culture medium into the fruiting bag with one end sealed by a bag filling machine, sealing the other end by a non-cotton cover body, sterilizing in a 0.14MPa autoclave for 3h, cooling to below 30 ℃, inoculating strains into the fruiting bag, and moving to a fermentation chamber with the temperature of 18-22 ℃ and the relative air humidity of 40-60%; after the bag is full of hypha, the fruiting bag is moved to a mushroom house, the temperature is 10-15 ℃, the relative humidity of air is 80-90%, the bag opening is opened, and the light induction is performed to promote the formation of mushroom buds; when the height of the sporocarp grows to 2-3cm, sheathing a plastic bag cylinder on the outside of the mushroom bag, continuing to grow until the height of the sporocarp grows to 20-25cm and the diameter of the pileus is 0.3-0.5cm, harvesting, screening the pileus with the color between light yellow and golden yellow, wherein the pileus is semicircular, the stipe is between white and light yellow, the fineness is fine, the base part has less villus, the pilus is not adhered, the browning is not easy to occur, and the yield is higher than that of the parent strain FHFHJ-12.
Performing monospore selfing on the screened binuclear bacterial strain FHJ-12 after a fruiting test, inoculating 2 monospore bacterial strains on a PDA culture medium flat plate at a distance of 1.5 +/-0.2 cm, culturing at the temperature of 22 +/-1 ℃, after hypha grows and contacts, taking mycelium at the contact part to be switched on a slant PDA culture medium, obtaining a binuclear selfing bacterial strain when the growth distance of the hypha reaches 1.5 +/-0.2 cm, performing hypha culture on the selfing bacterial strain, selecting a hybrid with dense hypha and high growth speed, and performing a fruiting test. Uniformly mixing 78g of cottonseed hulls, 15g of bran, 5g of corn meal and 2g of lime, adding 186g of water, and uniformly stirring to obtain a mushroom culture medium; adopting high-density polypropylene plastic with the specification of 17 multiplied by 33 multiplied by 0.05cm as a fruiting bag, adopting a bagging machine to fill a culture medium into the fruiting bag with one end sealed, sealing the other end with a non-cotton cover body, sterilizing in an autoclave with the pressure of 0.14MPa for 3h, then cooling to below 30 ℃, inoculating a strain into the fruiting bag, moving to a fermentation room, wherein the temperature of the fermentation room is 18-22 ℃, and the relative humidity of air is 40-60%; after the bag is full of hypha, the fruiting bag is moved to a mushroom house, the temperature is 10-15 ℃, the relative humidity of air is 80-90%, the bag opening is opened, and the light induction is performed to promote the formation of mushroom buds; when the height of the sporocarp grows to 2-3cm, sheathing a plastic bag cylinder on the outside of the pileus bag, continuing to grow until the height of the sporocarp grows to 20-25cm and the diameter of the pileus is 0.3-0.5cm, harvesting, screening the pileus with golden yellow color, wherein the pileus is semicircular, the stipe is between white and light yellow, the pilus at the base is few, the pilus is not adhesive and easy to brown, and the yield is higher than that of the strain FHFHJ-12-P46 of the initial strain.
Further fixing the strain FHJ-12-P46 by adopting a tissue separation method, selecting fresh fruit bodies after the fruiting test in the last step, taking flesh tissues at the joint of a pileus and a stipe by using a knife, transferring the flesh tissues to a new PDA culture medium under an aseptic condition for culture, obtaining pure hyphae after germination, namely a new strain, and finally selecting a new flammulina velutipes variety with excellent and stable inheritance properties, namely Jijin 12, through fruiting tests and property fixation in two cultivation seasons.
The cultivation method of the flammulina velutipes strain comprises the following steps:
(1) breeding a mother seed: inoculating Jijin 12 strain with diameter of 0.3-0.5cm to mother culture medium, culturing at 23 + -1 deg.C and air relative humidity of 50 + -5% in the dark for 6 days to obtain Jijin 12 mother strain; wherein the mass ratio of the Jijin 12 strain to the mother culture medium is 1: 100; a method for preparing mother culture medium comprises placing 200g peeled potato pieces in 600ml water, boiling for 30min, filtering, collecting filtrate, adding 400ml water, 20g glucose and 20g agar into the filtrate, diluting to 1000ml, heating with slow fire under stirring until the above components are completely dissolved, and sterilizing at 121 deg.C for 30 min.
(2) Stock culture: inoculating Jijin 12 mother strain in stock culture medium (stock culture medium filled into strain bag), and culturing at 23 + -1 deg.C and air relative humidity of 50 + -5% in dark condition for 32 days to obtain Jijin 12 stock strain;
wherein the mass ratio of the Jijin 12 mother strain to the stock culture medium is 4: 100; the raw material culture medium comprises 78g of cottonseed hull, 20g of bran, 1g of gypsum, 1g of lime and 186g of water, and the raw material culture medium is 1.05kg/cm 3 Keeping the pressure for 3h, and sterilizing by high-pressure steam for use;
taking 15cm multiplied by 26cm multiplied by 0.05cm (folded diameter multiplied by length multiplied by thickness) polypropylene plastics as strain bags, subpackaging by a bagging machine, wherein each strain bag can be filled with 350g of dry materials (namely, raw material culture medium is filled into the strain bag), mother seeds are propagated in the strain bag, and one bag of original seeds can be propagated as seeds to obtain 15 bags of cultivated species.
(3) Cultivating cultivars: inoculating Jijin 12 stock seed to a culture medium (the culture medium is filled into a strain bag), and culturing at 23 + -1 deg.C and air relative humidity of 50 + -5% in the dark for 37 days to obtain Jijin 12 cultivar; wherein the mass ratio of the Jijin 12 stock seeds to the culture medium of the cultivated species is 4: 100; the culture medium comprises 78g of cottonseed hull, 20g of bran, 1g of gypsum, 1g of lime and 186g of water;
the culture bag is made of 17cm × 36cm × 0.05cm (folded diameter × length × thickness) polypropylene plastic, and is filled with 500g dry weight (i.e., culture medium is filled into the culture bag), and 25 bags of cultivation seeds can be transferred to the culture bag 1.
(4) The fruiting bag is a high-density polyethylene plastic bag cylinder of 17cm multiplied by 33cm multiplied by 0.05cm, one end of the bag cylinder is tied by a plastic rope before bagging, the length of a film on the outer side of the tying is 2.2 +/-0.2 cm, a bag filling machine is adopted to fill a fruiting bag culture medium into the fruiting bag, 500g of dry materials can be filled into each bag on average, the other end of the bag is tied by the plastic rope after filling, after the fruiting bag culture medium is filled, after normal-pressure sterilization is carried out for 15 hours at 100 ℃, Jijin 12 cultivation seeds are inoculated on the fruiting bag culture medium according to the proportion of 6% by weight, inoculation is carried out by 2 persons in a matching way, in the inoculation, 1 person opens tying openings at two ends of the fruiting bag, and 1 person takes out a strain inoculating material surface from the inside of the strain bag by using an inoculation tool, the action is rapid, and the pollution chance is reduced. Wherein, the requirements of the cultivated species of Jijin 12 are as follows: the hypha of the strain is white and strong, and the age of the strain is within 15 days of the full bag; raw materials of the fruiting bag culture medium comprise 78g of cottonseed hulls, 15g of bran, 5g of corn flour, 2g of lime and 186g of water;
hypha culture: and (3) transferring fruiting bags containing inoculated Jijin 12 cultivars to a fungus growing room for hypha culture, transversely arranging the fruiting bags on a frame layer, controlling the temperature of the fungus growing room to be 18-22 ℃, ventilating, shading, preventing insects, cleaning and drying, controlling the relative humidity of air to be 50 +/-5%, checking bag by bag regularly, immediately processing after finding that mixed fungi are polluted, and preventing the mixed fungi from spreading.
And (3) fruiting management: after the cultured cultivars after hypha culture enter a bud forcing period, keeping the relative humidity of air at 85 +/-1%, the temperature of a mushroom shed at 11 +/-0.5 ℃, opening an air opening for a small time, inducing with low light to promote the formation of mushroom buds, and simultaneously mechanically scratching mushroom bags; when the buds grow into the mushroom bags and then enter a young mushroom period, adjusting the relative humidity of air to 90 +/-3 percent and the temperature of a mushroom shed to 16 +/-4 ℃, keeping the air fresh and in a low-light environment, and promoting primordial differentiation and branching; when young mushrooms grow to about 2cm, entering a mushroom inhibiting period, ventilating, reducing humidity and increasing light to inhibit growth of mushroom bodies, keeping for 3 days, promoting thickening of mushroom stems and mushroom caps, and keeping the mushroom bodies strong and tidy; after the mushroom inhibiting period is finished, spraying water in a proper amount to keep the relative humidity of air at 85 +/-3%, reducing ventilation, keeping the mushroom shed dark and promoting the extension of mushroom stems; the mechanical mushroom scratching step comprises the steps of opening a bag opening at one end of a mushroom bag, removing old mushroom blocks by using a hand rake, burning the hand rake on flame of an alcohol burner before use, and carrying out the same operation after the bag opening at the other end is subjected to first mushroom wetting.
When hypha cultivation and fruiting management are carried out, fruiting bags need to be placed on shelves of a semi-underground fruiting shed, the length of the fruiting shed is 80 meters, the width of the fruiting shed is 8 meters, the height of the fruiting shed is 2.2 meters, the middle of the fruiting shed is slightly arched, 0.4 meter is dug downwards, a plurality of shelf structures are arranged in the fruiting shed, the horizontal distance between every two adjacent shelf structures is 1m, the number of layers of each shelf structure is 5, the fruiting bags are placed on each layer, the distance between the bottom layer and the ground is 0.15-0.2m, the distance between every two adjacent layers is 0.4m, upper and lower ventilation openings are reserved on two side walls of the fruiting shed, the hole size is 0.3 x 0.3m, and the shed top is covered with a heat-insulating material.
Harvesting in due time: collecting when pileus diameter of herba Pteridis Davidii 12 fruiting body is 0.3-0.5cm, stipe diameter is 0.2-0.4cm, and stipe length is 20-25 cm; when the mushroom cultivation box is used for harvesting, the plastic bag opening is pressed by one hand, the mushroom is lightly held by the other hand to be pulled out, the base part of the stipe is cut to be neat by a small knife if the stipe is provided with a cultivation material, and then the stipe is flatly placed in a plastic basket to prevent the mushroom from being excessively loaded and crushing the mushroom body.
And (3) tide change management: jijin 12 can collect 3 tide mushrooms generally. The method comprises the following steps of harvesting a first tide of mushrooms, cleaning a material surface, raking old mushroom blocks and other impurities, binding a bag opening again, opening the bag opening at the other end, humidifying, inducing glimmer, entering fruiting period management again, weighing mushroom bags after harvesting every tide of mushrooms, enabling the weight of the mushroom bags to be lower than the original weight by 60%, supplementing water, enabling the water supplementing amount to be 80% of the water loss amount, and guaranteeing the fruiting yield of a second tide and a third tide.
The harvested needle mushroom has golden yellow pileus, bright color, average diameter of 0.25cm, yellow-white stipe, fine texture, less villi at the base of the stipe, no adhesion, and average content of lignin of the stipe of fruiting body at harvesting stage of 2.50 × 10 3 A 280nm The lignin content of/kg is significantly lower than that of the traditional variety (3.05 multiplied by 10) 3 A 280nm Per kg), the commercial mushroom has good palatability and is suitable for fresh eating; jijin 12 is suitable for an agricultural facility cultivation mode, the highest single-tide single-bag fruiting yield can reach 324.5g, three-tide mushrooms can be produced by a mushroom inter-tidal period water supplementing fruiting technology, the average total fruiting yield per bag can reach 720g, and the average total biological efficiency is 140%.
The method for measuring the lignin content comprises the following steps: selecting fresh needle mushroom fruiting body with complete mushroom body, no plant diseases and insect pests, no mechanical damage and seven-mature fruiting body, taking 2/3 parts of mushroom flesh tissue from mushroom stem to mushroom stem from top to bottom, adding 5ml of 95% ethanol, grinding, centrifuging for 7min, discarding supernatant, taking precipitate, washing with 95% ethanol for 3 times, washing with mixed solvent of ethanol and n-hexane at a volume ratio of 1:2 for 3 times, collecting precipitate, fully drying, dissolving the dried product in 25% bromoacetyl glacial acetic acid solution, keeping the temperature in a constant temperature water bath at 70 ℃ for 30min, adding 0.9ml of 2mol/L NaOH to terminate the reaction, adding 5ml of glacial acetic acid and 0.1ml of 7.5mol/L hydroxylamine hydrochloric acid, diluting to 10ml with glacial acetic acid, centrifuging for 7min, taking supernatant, measuring absorption value at 280nm, and expressing lignin content at 280nm of fresh weight, repeat 3 times, take the average as the lignin content of the stipe.
Example 2
The cultivation method of the flammulina velutipes strain Jijin 12 comprises the following steps:
(1) breeding a mother seed: inoculating Jijin 12 strain with diameter of 0.3-0.5cm to mother culture medium, culturing at 21 + -1 deg.C and air relative humidity of 45 + -5% in dark condition for 7 days to obtain Jijin 12 mother strain; wherein the mass ratio of the Jijin 12 strain to the mother culture medium is 1: 100; a method for preparing mother culture medium comprises placing 200g peeled potato pieces in 600ml water, boiling for 30min, filtering, collecting filtrate, adding 400ml water, 20g glucose and 20g agar into the filtrate, diluting to 1000ml, heating with slow fire under stirring until the above components are completely dissolved, and sterilizing at 121 deg.C for 30 min.
(2) Stock culture: inoculating Jijin 12 mother strain into stock culture medium (stock culture medium is filled into strain bag), culturing at 21 + -1 deg.C and air relative humidity of 55 + -5% in dark condition for 35 days to obtain Jijin 12 stock strain;
wherein the mass ratio of the Jijin 12 mother strain to the stock culture medium is 4: 100; the raw material culture medium comprises 78g of cottonseed hull, 20g of bran, 1g of gypsum, 1g of lime and 186g of water, and the raw material culture medium is 1.05kg/cm 3 Keeping the pressure for 3h, and sterilizing by high-pressure steam for use;
the culture medium is filled in 500ml/19ml (volume/caliber) glass bottles, a special cottonseed hull bottling machine is adopted for subpackaging, each bottle can be filled with 400g of dry materials (namely, the raw material culture medium is filled in the glass bottles), the stock seeds are propagated in the glass bottles, and 20 bottles of cultured seeds can be propagated in one bottle.
(3) Cultivating cultivars: inoculating Jijin 12 stock seed to a culture medium (the culture medium is filled into a strain bag), and culturing at 22 + -1 deg.C and air relative humidity of 55 + -5% in the dark for 37 days to obtain Jijin 12 cultivar; wherein the mass ratio of the Jijin 12 stock seeds to the culture medium of the cultivated species is 4: 100; the culture medium comprises 78g of cottonseed hull, 20g of bran, 1g of gypsum, 1g of lime and 186g of water;
the culture medium is contained in 500ml/19ml (volume/caliber) glass bottles, a special cottonseed hull bottling machine is adopted for subpackaging, each bottle can be filled with 400g of dry materials (namely, the glass bottles are filled with the culture medium of the cultivated species), the original species are propagated in the glass bottles, and 20 bags of fruiting fungus bags can be inoculated in one bottle of cultivated species.
(4) The fruiting bag is made of 17cm × 33cm × 0.05cm high-density polyethylene plastic, one end of a bag cylinder needs to be tied up by a plastic rope before bagging, the length of a tied-up outer side film is 2.2 +/-0.2 cm, a bag filling machine is used for filling a fruiting bag culture medium into the fruiting bag, the other end of the bag is tied up by the plastic rope after filling, after the fruiting bag culture medium is filled, sterilization is carried out at 100 ℃ for 15 hours under normal pressure, Jijin 12 cultivation seeds are inoculated on the fruiting bag culture medium according to the weight percentage of 8%, inoculation is carried out by 2 persons in a matching mode, tying openings at two ends of the fruiting bag are opened by 1 person, in an inoculation box, the 1 person takes out a strain inoculating material surface from the strain bag by using an inoculation tool, the action is fast, and the pollution opportunity is reduced. Wherein, the requirements of the cultivated species of Jijin 12 are as follows: the hypha of the strain is white and strong, and the age of the strain is within 15 days of the full bag; raw materials of the fruiting bag culture medium comprise 78g of cottonseed hulls, 15g of bran, 5g of corn flour, 2g of lime and 186g of water;
hypha culture: the fruiting bag containing inoculated Jijin 12 cultivars is moved to a spawn running room for hypha culture, the fruiting bags are horizontally arranged on a frame layer, the spawn running room is required to be constant in temperature (21 +/-0.5 ℃), ventilated, protected from light, protected from insects, clean and dry, the relative humidity of air is 45 +/-5%, the bag-by-bag inspection is carried out regularly, and the fruiting bag is immediately treated after the pollution of mixed bacteria is found to prevent the diffusion and spread.
And (3) fruiting management: after the cultured cultivars with hypha enter a bud forcing period, keeping the relative humidity of air at 90 percent and the temperature of a mushroom shed at 11 +/-0.5 ℃, opening an air opening for inducing with low light to promote the formation of mushroom buds, and simultaneously mechanically scratching mushroom bags; when the mushroom buds grow into the mushroom bags, the relative humidity of the air is adjusted to 87 +/-3%, the temperature of the mushroom shed is 13 +/-2 ℃, the air is kept fresh and in a low-light environment, and primordial differentiation and branching are promoted; when the young mushrooms grow to about 2cm, entering a mushroom inhibiting period, ventilating, and irradiating with moisture and light to inhibit growth of mushroom bodies for 3 days, so as to promote thickening of mushroom stems and mushroom caps, and make the mushroom bodies robust and neat; after the mushroom inhibiting period is finished, spraying water in a proper amount to keep the relative humidity of air at 85 +/-3%, reducing ventilation, keeping the mushroom shed dark and promoting the extension of mushroom stems; the mechanical mushroom scratching step comprises the steps of opening a bag opening at one end of a mushroom bag, removing old mushroom blocks by using a hand rake, burning the hand rake on flame of an alcohol burner before use, and carrying out the same operation after the bag opening at the other end is subjected to first mushroom wetting.
When hypha cultivation and fruiting management are carried out, fruiting bags need to be placed in a semi-underground fruiting shed, the length of the fruiting shed is 80 meters, the width of the fruiting shed is 8 meters, the height of the fruiting shed is 2.2 meters, the middle of the fruiting shed is slightly arched, the fruiting shed is dug downwards for 0.4 meter, a plurality of shelf structures are arranged in the fruiting shed, the horizontal distance between every two adjacent shelf structures is 1m, the number of layers of each shelf structure is 5, the fruiting bags are placed on each shelf, the distance between the bottom layer and the ground is 0.15-0.2m, the distance between the two adjacent shelf layers is 0.4m, upper and lower ventilation openings are reserved on the two side walls of the fruiting shed, the hole size is 0.3 x 0.3m, and the shed top is covered with heat-insulating materials.
Harvesting in due time: collecting when pileus diameter of herba Pteridis Davidii 12 fruiting body is 0.3-0.5cm, stipe diameter is 0.2-0.4cm, and stipe length is 20-25 cm; when the mushroom cultivation box is used for harvesting, the plastic bag opening is pressed by one hand, the mushroom is lightly held by the other hand to be pulled out, the base part of the stipe is cut to be neat by a small knife if the stipe is provided with a cultivation material, and then the stipe is flatly placed in a plastic basket to prevent the mushroom from being excessively loaded and crushing the mushroom body.
And (3) tide change management: jijin 12 can collect 3 tide mushrooms generally. After harvesting the first tide of mushrooms, cleaning the material surface, raking old mushroom blocks and other impurities, binding the bag opening again, opening the bag opening at the other end, humidifying, inducing by low light level, entering the fruiting period management again, weighing the mushroom bags after harvesting every tide of mushrooms, wherein the weight of the mushroom bags is lower than 60% of the original weight, replenishing water, the amount of the replenished water is 75% of the water loss amount, and the step can guarantee the fruiting output of the second tide and the third tide.
The harvested needle mushroom has golden yellow pileus, bright color, average diameter of 0.25cm, yellow-white stipe, fine texture, less villi at the base of the stipe, no adhesion, and average content of stipe lignin of fruiting body of harvesting period of 2.45 × 10 3 A 280nm The lignin content of/kg is significantly lower than that of the traditional variety (3.05 multiplied by 10) 3 A 280nm Per kg), the commercial mushroom has good palatability and is suitable for fresh eating; jijin 12 is suitable for agricultural facility cultivation mode, the highest yield of single-head tide single-bag fruiting can reach 290g, and fruiting can be realized by supplementing water during the inter-tidal periodBy adopting the technology, three-tide mushrooms can be produced, the average total yield per bag of mushrooms can reach 645g, and the average total biological efficiency is 129%.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications of the invention may be made without departing from the spirit or scope of the invention.
Claims (11)
1. Golden mushroom (Flammulina filiformis) strain Jijin 12 is characterized in that the preservation number is CGMCC No. 19644.
2. A method for cultivating a Flammulina velutipes strain according to claim 1, comprising the steps of,
(1) and (3) parent seed propagation: inoculating Jijin 12 to a mother strain culture medium, and performing first culture to obtain Jijin 12 mother strains;
(2) stock culture: inoculating the Jijin 12 mother seeds on an original seed culture medium, and performing second culture to obtain Jijin 12 original seeds;
(3) cultivating cultivars: inoculating the Jijin 12 stock seed to a cultivated species culture medium, and performing third culture to obtain a Jijin 12 cultivated species;
(4) inoculating the Jijin 12 cultivar into a fruiting bag culture medium, and harvesting after hypha culture and fruiting management.
3. The cultivation method according to claim 2, wherein in the step (2), the stock culture medium comprises the following raw materials in a mass ratio of (75-80): (17-21): (0.5-1.5): (0.7-1.3) cottonseed hulls, bran, gypsum and lime, wherein the mass ratio of dry materials to water in the stock culture medium is 1: (1.8-2.3);
in the step (3), the culture medium of the cultivar comprises the following raw materials in a mass ratio of (75-80): (17-21): (0.5-1.5): (0.7-1.3) cottonseed hulls, bran, gypsum and lime, wherein the mass ratio of dry materials to water in the culture medium of the cultivated species is 1: (1.8-2.3).
4. The cultivation method according to claim 2 or 3, wherein the specific step of fruiting management comprises,
the fruiting bag after mycelium culture enters a bud forcing period, and mushroom buds are promoted to form under the conditions of air relative humidity of 80-90%, temperature of 10-12 ℃, ventilation and low light;
when the mushroom buds grow into the mushroom bags, the mushroom buds enter a young mushroom period and grow under the conditions that the relative humidity of air is 85-95%, the temperature is 12-20 ℃ and ventilation is performed;
when the young mushroom grows to 2 +/-0.5 cm, entering a mushroom inhibiting period, and growing under the conditions that the relative humidity of air is 80-85%, the temperature is 12-18 ℃ and ventilation is performed;
after the mushroom inhibiting period, the mushroom is grown under the condition that the relative humidity of air is 83-86%.
5. The cultivation method according to claim 2 or 3, wherein in the step (4), the raw materials of the fruiting bag culture medium comprise the following components in a mass ratio of (75-80): (13-17): (4-6): (1-3) cottonseed hulls, bran, corn meal and lime, wherein the mass ratio of dry materials to water in the fruiting bag culture medium is 1: (1.8-2.3);
the mass concentration of water in the fruiting bag culture medium is 60-70%;
when the fruiting bag culture medium is inoculated, the fruiting bag culture medium is placed in a fungus bag with openings at two ends, the fungus bag is sterilized at 100 ℃ for 12-18h, and inoculation is carried out at two ends of the fungus bag after the fungus bag is cooled to room temperature.
6. The cultivation method according to claim 2 or 3, wherein the hypha cultivation is performed under conditions of a temperature of 18 to 22 ℃ and a relative air humidity of 40 to 60%.
7. The cultivation method according to claim 2 or 3, wherein the hypha cultivation and the fruiting management are performed in a semi-underground fruiting shed;
a plurality of shelf structures are arranged in the fruiting shed;
the ratio of the length, the width and the height of the fruiting shed is 80: 8: (2-3).
8. The cultivation method as claimed in claim 2, wherein in step (1), the raw materials of the mother culture medium comprise (180-220) by mass: (19-22): (18-21) peeled potatoes, glucose and agar; the first culture temperature is 20-25 deg.C, air relative humidity is 40-60%, and the time is 5-7 days; the mass ratio of the Jijin 12 to the mother culture medium is (0.8-1.2): 100, respectively;
in the step (2), the temperature of the second culture is 20-25 ℃, the relative humidity of air is 40-60%, and the time is 30-35 days; the mass ratio of the Jijin 12 mother seeds to the stock seed culture medium is (3-5): 100;
in the step (3), the temperature of the third culture is 20-25 ℃, the relative humidity of air is 40-60%, and the time is 35-40 days; the mass ratio of the Jijin 12 stock seeds to the culture medium of the cultivated species is (3-5): 100.
9. the cultivation method as claimed in claim 8, wherein the raw materials of the mother culture medium include 200 parts by weight of peeled potatoes, 20 parts by weight of glucose and 20 parts by weight of agar.
10. The cultivation method as claimed in claim 2, 3 or 9, wherein in the step (4), harvesting is performed when the cap diameter of the flammulina velutipes is 0.3-0.5cm, the stalk diameter is 0.2-0.4cm, and the length is 20-25 cm.
11. The cultivation method as claimed in claim 2, 3 or 9, further comprising the operation step of tide change management after the harvest;
the number of times of tide change management is 2-3;
after every tide of mushrooms is harvested, water is supplemented, and the water supplementing amount is 75-85% of the water loss amount.
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