CN107034147B - Breeding method and cultivation method for industrial bottle cultivation oyster mushroom variety - Google Patents
Breeding method and cultivation method for industrial bottle cultivation oyster mushroom variety Download PDFInfo
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Abstract
The invention discloses a breeding method for an industrial bottle cultivation oyster mushroom variety, which takes a high-yield strain P99 and a wild strain NY-2 as parent strains, creates a batch of new oyster mushroom germplasm by a monospore hybridization technology, adopts an industrial bottle cultivation fruiting mode for circular planting, screens out an excellent strain suitable for industrial production: the strain is black, has no stalk, has thick mushroom cap, good toughness, is transport-resistant, has the mushroom growing temperature of 12-15 ℃, the mushroom growing uniformity of more than 95 percent, the average yield of a single bottle of 180g, the weight of a single bottle of culture material of 250g, the highest yield of 220 g/bottle, the biological efficiency of a batch of mushrooms of more than 70 percent, the production period of 40 days, no plant diseases and insect pests in the production period, can realize annual supply, and fills the blank of domestic factory-like special varieties of the oyster mushrooms.
Description
Technical Field
The invention relates to cultivation of oyster mushrooms, in particular to a breeding method for an industrial bottle cultivation oyster mushroom variety, and further relates to an industrial cultivation method for the oyster mushrooms.
Background
The oyster mushroom has become a necessary dish on the public dining table due to its rich nutrition and moderate price. The total yield of pleurotus ostreatus in 2015 in China reaches 590.18 ten thousand tons, and the pleurotus ostreatus is the third major edible fungus which can be artificially planted. Henan province is the great province of oyster mushroom production, and the total yield accounts for 30% of the total amount of oyster mushrooms in China. The most suitable fruiting temperature of the oyster mushrooms is 12-18 ℃, the oyster mushrooms are influenced by environmental climate, the oyster mushroom production in Henan province is mostly concentrated in three seasons of autumn, winter and spring, the fruiting difficulty can occur when the temperature is lower than 4 ℃ in winter or the temperature is higher than 25 ℃ in summer, and the continuous supply of the market all year round can not be ensured. Meanwhile, the production mode of the traditional oyster mushroom is low in mechanization degree, more in physical labor, high in labor price and the like, and the planting profit of the oyster mushroom is directly reduced. The industrial production of edible fungi has rapidly developed and grown all over the country due to the adoption of stereoscopic planting, high production efficiency, high automation level, stable product quality and other advantages, and the number of edible fungi factory production enterprises in the country reaches more than 700 at present. With the maturity of the industrialized production technology of flammulina velutipes and pleurotus eryngii, the market gradually becomes saturated, so that the industrialized production of pleurotus ostreatus becomes the development direction of the next step. Because the traditional oyster mushroom varieties require 3-4 times of fruiting in production and the biological efficiency of industrialized production needs to be over 70 percent, the oyster mushroom varieties in the traditional cultivation mode are directly selected, the defects of low fruiting uniformity, low yield and biological efficiency of one-time mushroom, carbon dioxide intolerance and the like exist, the existing bottle cultivation production mode cannot be adapted, and a new oyster mushroom new variety suitable for industrialized bottle cultivation is urgently needed to be cultivated.
Disclosure of Invention
The invention aims to provide a breeding method for industrially bottle-cultured oyster mushroom varieties, and simultaneously provides an industrially cultivation method for the bred oyster mushroom varieties.
In order to achieve the purpose, the invention can adopt the following technical scheme:
the breeding method for the industrial bottle cultivation oyster mushroom variety comprises the following steps:
firstly, selecting a conventionally examined strain P99 and a wild strain NY-2 as parent strains;
secondly, respectively separating 20 monospores from the examined strain P99 and the wild strain NY-2, respectively selecting 6 representative monospores according to the difference of morphological characteristics of hyphae among the monospores, then hybridizing two by two, performing plate culture by adopting a PDA (personal digital assistant) culture medium, selecting hybrid strains at two sides of an antagonistic line after one week, respectively transferring the hybrid strains to a new culture dish for culture, and numbering; identifying the specificity between the hybrid strains and parent strains by adopting an antagonism method, and determining the hybrid strains with the specificity as test strains;
thirdly, placing the determined test strain into a cotton seed hull clinker small bag for cultivation, and performing a fruiting test: observing the uniformity of the fruiting, the shape of the fruiting body and the yield of the first-batch mushrooms; screening out strains with biological efficiency of more than 50 percent, black color, short stalk and thick pileus of a first crop of mushrooms as first round bottle cultivation test strains;
fourthly, preparing culture materials according to the proportion of 88 percent of cottonseed hulls, 10 percent of bran and 2 percent of lime, and keeping the water content at 61 percent; adopting 1000ml strain bottles as containers, filling 250g of dry materials in each bottle, inoculating 4 baskets of strains, inoculating 16 bottles/basket of strains, culturing at 22-24 ℃, filling the bottles with mycelia for 30 days, generating buds after 5-7 days of after-ripening, moving the strains into a fruiting chamber, removing a pileus, keeping the indoor temperature at 15-18 ℃, the humidity at 80-90%, controlling the concentration of carbon dioxide below 600ppm, illuminating 400-500 lux, harvesting 7-8 ripe fruiting bodies, observing fruiting uniformity, fruiting properties, weighing yield, calculating biological efficiency, and screening 1 excellent hybrid strain as a 2 nd round bottle cultivation test variety;
taking the excellent strains obtained by screening as test strains, taking the existing three high-yield strains and parent strains P99 as control strains, planting 10 baskets of each variety, culturing 20 bottles in each basket in the fourth step, randomly extracting 3 baskets of each variety as weighing samples, observing the uniformity of fruiting, the properties of fruiting bodies, recording the yield of each bottle and the number of the differentiated fruiting bodies, and calculating the biological efficiency; after the fresh mushrooms are harvested, the fresh mushrooms are dried at 50 ℃, 100g of dry products are respectively weighed for each variety, the quality of each variety is analyzed, the superiority and inferiority of the strain are checked, and the oyster mushroom varieties suitable for industrial bottle cultivation can be determined by the indexes superior to or equal to those of the high-yield strain and the parent strain P99.
The industrial cultivation method of the oyster mushroom variety bred by the invention comprises the following steps:
firstly, preparing culture material for inoculating bottle
Weighing 33-68% of cottonseed hulls, 20-50% of corncobs, 5% of bean pulp, 5-10% of bran and 2% of lime by weight percent, uniformly mixing the materials, filling the mixture into a 1100 ml polypropylene plastic bottle, wherein each bottle weighs 760 + 10g, and sterilizing the mixture in an aseptic inoculation chamber after high-pressure sterilization;
second step, inoculation
When the temperature of the inoculation bottle is reduced to 40 ℃, starting inoculation, adopting the selected solid strain with the inoculation amount of 10 percent, sealing the material surface after inoculation, and moving the material surface into a culture room;
the third step, culturing
Keeping the temperature of the culture room at 22-25 ℃, and culturing in a dark place; after the bottle is full of hypha in 28-29 days, reducing the temperature of the culture room to 15-18 ℃, keeping for 3 days, starting bud formation in the strain bottle, removing the bottle cap, and entering a fruiting room; a three-dimensional oblique-swinging fruiting mode is adopted, the temperature of a fruiting chamber is kept at 12-15 ℃, the carbon dioxide concentration is below 600ppm, and the illumination intensity is 200 lux; and harvesting the fruiting body 7 after six days.
In actual preparation, the formula of the culture material of the inoculation bottle is as follows: 68% of cottonseed hulls, 20% of corncobs, 5% of soybean meal, 5% of bran and 2% of lime.
The formula of the culture material of the inoculation bottle can be as follows: 58% of cottonseed hulls, 30% of corncobs, 5% of soybean meal, 5% of bran and 2% of lime.
The formula of the culture material of the inoculation bottle can also be as follows: 43% of cottonseed hulls, 40% of corncobs, 5% of soybean meal, 10% of bran and 2% of lime.
The formula of the culture material of the inoculation bottle can also be as follows: 33% of cottonseed hulls, 50% of corncobs, 5% of soybean meal, 10% of bran and 2% of lime.
The invention has the advantages that the conventionally approved variety P99 and the wild strain NY-2 are adopted as parent strains, the new germplasm of the oyster mushroom is bred by a monospore hybridization technology, and an excellent strain suitable for industrial production is screened out: P14Y6, the strain is black, has no handle, thick pileus, good toughness, transportation resistance, fruiting temperature of 12-15 ℃, fruiting uniformity of more than 95%, average yield of 180g in a single bottle, and biological efficiency of a batch of mushrooms of more than 70%; the selected oyster mushroom variety is circularly planted in an industrial bottle cultivation fruiting mode, the production period is 40 days, no plant diseases and insect pests occur in the production period, annual supply can be realized, and the blank of domestic special industrial oyster mushroom varieties is filled.
Drawings
FIG. 1 is a photograph of the fruiting site of the Pleurotus ostreatus strain P14Y6 screened by the present invention.
FIG. 2 is a photograph of a fruit body of Pleurotus ostreatus strain P14Y6 selected by the present invention.
Detailed Description
The breeding method for the industrial bottle cultivation oyster mushroom variety comprises the following steps:
firstly, selecting a conventionally approved strain P99 (called P for short, introduced from the institute of agricultural resources and division of Chinese agricultural academy of sciences), and a wild strain NY-2 (called Y for short, provided by agricultural academy of Shanghai city, Henan province) as parent strains, wherein the biological characteristics of the strain are that the wild isolated strain is dark gray, has no handle, and has strong resistance, low yield and supernatural;
secondly, separating 20 single cells from the examined strain P99 with the serial numbers of P1, P2, … … and P20; similarly, 20 monospores are separated from the wild strain NY-2, and the numbers of the monospores are Y1, Y2, … … and Y20 in sequence; respectively selecting 6 representative monospores according to different morphological characteristics of hyphae among the monospores, hybridizing in pairs, performing plate culture by adopting a PDA (PDA) culture medium, selecting hybrid strains at two sides of an antagonistic line after one week, respectively transferring the hybrid strains to a new culture dish for culture, obtaining 72 hybrid combinations, wherein the numbers of the hybrid combinations are respectively test strains, the following table 1 is six monospores respectively selected from a strain P99 and a strain NY-2, and the table 2 is the number of the hybrid combinations obtained after the selected monospores are hybridized in pairs;
identifying the specificity among hybrid strains and between the hybrid strains and parent strains by adopting an antagonism method, and screening 49 hybrid strains with specificity to determine as test strains;
thirdly, placing the determined test strain into a cotton seed hull clinker small bag for cultivation, and performing a fruiting test: 10 bags are arranged in each group, each bag is 200g of dry material, mushroom fruiting management is carried out conventionally, and the differences of yield and biological characters among different hybrid strains are weighed and recorded; after the uniformity, the shape and the yield of fruiting bodies are observed, 15 strains with biological efficiency of more than 50 percent, black (gray), short stalk and thicker pileus and yield of more than 700g are screened out: P14Y6, P14Y9, Y9P9, Y9P7, Y9P10, Y6P10, P3Y6, Y6P3, Y6P9, Y6P14, Y9P14, P10Y9, Y9P3, Y8P3, P10Y6, see table 3, as first round of bottle cultivation test strains;
fourthly, preparing culture materials according to the proportion of 88 percent of cottonseed hulls, 10 percent of bran and 2 percent of lime, and keeping the water content at 61 percent; the method comprises the following steps of adopting 1000ml strain bottles as containers, inoculating 250g of dry materials in each bottle, inoculating 15 hybrid strains in each bottle for 4 baskets, 16 bottles/basket, culturing at 22-24 ℃, enabling mycelia to fill the bottles after 30 days, enabling buds to appear after 5-7 days of after-ripening, moving the hybrid strains into a fruiting chamber, removing mushroom caps, keeping the indoor temperature at 15-18 ℃, humidity at 80-90%, controlling the concentration of carbon dioxide below 600ppm, illuminating at 400-500 lux, harvesting 7-8 mature fruiting bodies, observing fruiting uniformity, fruiting body properties, weighing yield and calculating biological efficiency, wherein the P14Y6 strain is optimal in performance, and the results of the P14Y6 strain are analyzed and concentrated, and the average fruiting rate is 93.7%; the yield of one crop of mushrooms in each bottle reaches 178g on average, the biological efficiency is 71.2%, the yield is highest, the shape of a fruiting body is good, the pileus is blue and black, the diameter is 3-4.5 cm, and the thickness is 1-1.2 cm; the toughness is strong; the stipe is short, 1-1.2 cm long and 0.5-1 cm in diameter, and is shown in Table 4 below;
taking the excellent strain P14Y6 obtained by screening as a reference strain, taking the existing three high-yield strains (Heizheng No. 650, nongping No. 4 and Huimei No. 2) and the parent strain P99 as control strains, planting 10 baskets of each strain and 20 bottles of each basket, and cultivating in a fourth step, wherein the excellent strains can be seen in the following tables 5 and 6: the hyphae of the 5 test strains grow in a full bottle in 27-29 days, and the difference of the bottle filling time is small; the uniformity of the fruiting rate of P14Y6 is 96.8%, which is obviously higher than that of the parent strain and 3 control strains; the average single-bottle yield is 185g, the biological efficiency is 74 percent, the yield is obviously higher than that of 4 control varieties by 10-14 percent, the maximum weight of a single plant reaches 220 g/bottle, and the biological efficiency is 88 percent at the moment. The characteristics of the fruiting bodies show that the P14Y6 mushroom buds are less in differentiation, the number of grown mushrooms is large, the size of pileus is moderate, the stipe is short, and the toughness is strong; the three control varieties of black rice 650, nongping No. 4 and Huimei No. 2 are more differentiated, the number of grown mushrooms is small, and the toughness of pileus is poor. Weighing a batch of quantitative fresh mushrooms after the fresh mushrooms are harvested, drying the fresh mushrooms at 50 ℃, weighing 100g of dry products of each variety according to a quartering method, analyzing the quality of the dry products, checking the superiority and inferiority of the strains,
from the quality analysis result of the fruiting body, the P14Y6 strain contains 17 amino acids, the total amino acid content of each 100g of dry mushroom is 17.12g, the protein content is 25g, and is slightly lower than that of the parent strain; the crude fat content is 1.6g, which is equivalent to that of the parent; 7.72g of crude fiber is slightly higher than that of the parent; the protein content is higher than 1.3-3.4% of 3 control strains, the crude fiber content is higher than that of the control strains, and the difference between the total amino acid content and the crude fat content is small, which is shown in tables 7 and 8.
The indexes of the screened strain P14Y6 are all superior to or equal to those of the existing high-yield strain and parent strain, and the strain P14Y6 can be determined to be a new oyster mushroom variety suitable for industrial bottle cultivation.
The industrial cultivation method of the screened oyster mushroom variety P14Y6 comprises the following steps:
firstly, preparing culture material for inoculating bottle
In order to reduce the production cost, the invention adopts four culture material formulas consisting of cottonseed hulls, corncobs, bean pulp, bran and lime according to the following weight percentage, and specifically comprises the following components:
formula 1: 68% of cottonseed hulls, 20% of corncobs, 5% of soybean meal, 5% of bran and 2% of lime;
and (2) formula: 58% of cottonseed hulls, 30% of corncobs, 5% of soybean meal, 5% of bran and 2% of lime;
and (3) formula: 43% of cottonseed hulls, 40% of corncobs, 5% of soybean meal, 10% of bran and 2% of lime;
and (4) formula: 33% of cottonseed hulls, 50% of corncobs, 5% of soybean meal, 10% of bran and 2% of lime;
accurately weighing the raw materials, pre-wetting the corncobs according to a material-water ratio of 1:2.2 18-24 hours in advance, adding other raw materials, adding water according to a material-water ratio of 1:1.3, stirring for 40 minutes by using a stirrer, connecting the stirrer with an automatic bottling machine, filling 760 + 10g of each bottle into a 1100 ml polypropylene plastic bottle, autoclaving for 120 minutes, taking out the bottle after the pressure is reduced to 0, entering a sterile inoculation chamber for sterilization, installing air purification sterile inoculation equipment in the sterile inoculation chamber, and sterilizing for 40 minutes by using ultraviolet rays and ozone;
second step, inoculation
Opening an air purification filter, starting inoculation when the temperature of a strain bottle is reduced to about 40 ℃, inoculating 10 percent of solid strain P14Y6 selected by the invention, sealing the surface of the material after inoculation, and moving the material into a culture room;
the third step, culturing
Keeping the temperature of the culture room at 22-25 ℃, and culturing in a dark place; after the bottle is full of hypha in 28-29 days, reducing the temperature of the culture room to 15-18 ℃, keeping for 3 days, starting bud formation in the strain bottle, removing the bottle cap, and entering a fruiting room; a three-dimensional inclined-swinging mushroom growing mode is adopted, and an air exhaust system, an atomization and humidification system, an LED illumination system, an air conditioning system, an environment monitoring system, a hoisting machine and an automatic control system are installed in a mushroom growing chamber. The mushroom is grown out by adopting a three-dimensional inclined shelf in a room, the shelf is flexibly moved into one place, 3 baskets of strain bottles can be placed on two sides of the shelf respectively, and the basket and the shelf form an angle of 60 degrees. Directly conveying and placing the fruiting frame placed outdoors on a fruiting frame by adopting a hoisting machine, wherein the area of the fruiting chamber is 150m28000 bottles can be placed at one time, the indoor temperature is kept at 12-15 ℃, the carbon dioxide concentration is below 600ppm, and the illumination intensity is above 200 Lux. After the buds appear, the fruiting bodies grow to 7 days, and are picked up after being mature. The picked strain bottles are directly moved out of the fruiting chamber by a hoisting machine, are directly transported to a bottle digging machine, are dug, and can be directly bottled again to realize cyclic utilization.
Table 9 shows comparative data for each index under the same management conditions when different formulations are used for the compost (pure cotton seed hulls are used as compost control formulation CK):
as can be seen from the results shown in table 9: the hypha growth vigor, the yield and the input-output ratio of the culture material formulas 1-4 adopted by the invention are superior to those of pure cotton seed hulls serving as the culture material, and the input-output ratio is the highest and is 1:2.5 when the formula 3 (43% of the cotton seed hulls, 40% of corn cobs, 5% of bean pulp, 10% of bran and 2% of lime) is adopted as the culture material of the oyster mushroom strains.
Claims (6)
1. A breeding method for an industrial bottle cultivation oyster mushroom variety is characterized by comprising the following steps: the method comprises the following steps:
firstly, selecting an examined strain P99 and a wild strain NY-2 as parent strains;
secondly, respectively separating 20 monospores from the examined strain P99 and the wild strain NY-2, respectively selecting 6 representative monospores according to the difference of morphological characteristics of hyphae among the monospores, then hybridizing two by two, performing plate culture by adopting a PDA (personal digital assistant) culture medium, selecting hybrid strains at two sides of an antagonistic line after one week, respectively transferring the hybrid strains to a new culture dish for culture, and numbering; identifying the specificity between the hybrid strains and parent strains by adopting an antagonism method, and determining the hybrid strains with the specificity as test strains;
thirdly, placing the determined test strain into a cotton seed hull clinker small bag for cultivation, and performing a fruiting test: observing the uniformity of the fruiting, the shape of the fruiting body and the yield of the first-batch mushrooms; screening out strains with biological efficiency of more than 50 percent, black color, short stalk and thick pileus of a first crop of mushrooms as first round bottle cultivation test strains;
fourthly, preparing culture materials according to the proportion of 88 percent of cottonseed hulls, 10 percent of bran and 2 percent of lime, and keeping the water content at 61 percent; adopting 1000ml strain bottles as containers, filling 250g of dry materials in each bottle, inoculating 4 baskets of strains, inoculating 16 bottles/basket of strains, culturing at 22-24 ℃, filling the bottles with mycelia for 30 days, generating buds after 5-7 days of after-ripening, moving the strains into a fruiting chamber, removing a pileus, keeping the indoor temperature at 15-18 ℃, the humidity at 80-90%, controlling the concentration of carbon dioxide below 600ppm, illuminating 400-500 lux, harvesting 7-8 ripe fruiting bodies, observing fruiting uniformity, fruiting properties, weighing yield, calculating biological efficiency, and screening 1 excellent hybrid strain as a 2 nd round bottle cultivation test variety;
taking the excellent strains obtained by screening as test strains, taking the existing three high-yield strains and parent strains P99 as control strains, planting 10 baskets of each variety, culturing 20 bottles in each basket in the fourth step, randomly extracting 3 baskets of each variety as weighing samples, observing the uniformity of fruiting, the properties of fruiting bodies, recording the yield of each bottle and the number of the differentiated fruiting bodies, and calculating the biological efficiency; after the fresh mushrooms are harvested, the fresh mushrooms are dried at 50 ℃, 100g of dry products are respectively weighed for each variety, the quality of each variety is analyzed, the superiority and inferiority of the strain are checked, and the oyster mushroom varieties suitable for industrial bottle cultivation can be determined by the indexes superior to or equal to those of the high-yield strain and the parent strain P99.
2. An industrial cultivation method of an oyster mushroom variety according to claim 1, characterized in that: the method comprises the following steps:
firstly, preparing culture material for inoculating bottle
Weighing 33-68% of cottonseed hulls, 20-50% of corncobs, 5% of bean pulp, 5-10% of bran and 2% of lime by weight percent, uniformly mixing the materials, filling the mixture into a 1100 ml polypropylene plastic bottle, wherein each bottle weighs 760 + 10g, and sterilizing the mixture in an aseptic inoculation chamber after high-pressure sterilization;
second step, inoculation
When the temperature of the inoculation bottle is reduced to 40 ℃, starting inoculation, adopting the selected solid strain with the inoculation amount of 10 percent, sealing the material surface after inoculation, and moving the material surface into a culture room;
the third step, culturing
Keeping the temperature of the culture room at 22-25 ℃, and culturing in a dark place; after the bottle is full of hypha in 28-29 days, reducing the temperature of the culture room to 15-18 ℃, keeping for 3 days, starting bud formation in the strain bottle, removing the bottle cap, and entering a fruiting room; a three-dimensional oblique-swinging fruiting mode is adopted, the temperature of a fruiting chamber is kept at 12-15 ℃, the carbon dioxide concentration is below 600ppm, and the illumination intensity is 200 lux; and harvesting the fruiting body 7 after six days.
3. The industrial cultivation method of an oyster mushroom variety according to claim 2, comprising: the formula of the culture material of the inoculation bottle is as follows: 68% of cottonseed hulls, 20% of corncobs, 5% of soybean meal, 5% of bran and 2% of lime.
4. The industrial cultivation method of an oyster mushroom variety according to claim 2, comprising: the formula of the culture material of the inoculation bottle is as follows: 58% of cottonseed hulls, 30% of corncobs, 5% of soybean meal, 5% of bran and 2% of lime.
5. The industrial cultivation method of an oyster mushroom variety according to claim 2, comprising: the formula of the culture material of the inoculation bottle is as follows: 43% of cottonseed hulls, 40% of corncobs, 5% of soybean meal, 10% of bran and 2% of lime.
6. The industrial cultivation method of an oyster mushroom variety according to claim 2, comprising: the formula of the culture material of the inoculation bottle is as follows: 33% of cottonseed hulls, 50% of corncobs, 5% of soybean meal, 10% of bran and 2% of lime.
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| CN108739045A (en) * | 2018-03-26 | 2018-11-06 | 四川省农业科学院土壤肥料研究所 | No. 7 new strains of pleurotus cornucopiae and its selection and breeding and propagation method |
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| CN113475309A (en) * | 2021-08-05 | 2021-10-08 | 河南省农业科学院植物营养与资源环境研究所 | Method for controlling trichoderma pollution of bailing mushroom fungus bottles or fungus bags produced in factory |
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