CN108048346B - Compound microbial inoculum for morchella cultivation and external nutrition and application thereof - Google Patents
Compound microbial inoculum for morchella cultivation and external nutrition and application thereof Download PDFInfo
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Abstract
The invention discloses a compound microbial inoculum for external nutrition of morchella cultivation and application thereof, belonging to the field of edible mushroom cultivation. The composite microbial inoculum used by the nutrient material comprises cellulomonas flavigena, candida utilis and geotrichum candidum. Adding a compound microbial inoculum according to a formula of 30-35 parts of rice hulls, 30-35 parts of straws, 30-35 parts of mushroom bran, 1-2 parts of gypsum, 2-3 parts of glucose, 1-2 parts of salt and 0.1-0.3 part of monopotassium phosphate for fermentation, filling the mixture into a plastic mushroom bag, perforating the same side of the material bag, arranging 2 lines on two sides of a morchella esculenta mushroom bed, and tightly adhering and covering soil at the perforated part, wherein the aperture is 1-2 cm. The complex microbial inoculum of the invention has symbiotic coordination without mutual antagonism, and can provide sufficient external nutrition after being fermented with the substrate. The invention utilizes the compound microbial inoculum and the mushroom bran rich in growth promoting substances as external nutrition, and can obviously improve the yield and accelerate the germination speed of the morchella esculenta.
Description
Technical Field
The invention belongs to the field of edible fungus cultivation and microorganism utilization, and particularly relates to a morchella cultivation externally-applied nutrient composite microbial inoculum and a nutrient material prepared by using the same.
Background
Morchella esculenta (L.) Pers is a rare edible fungus and medicinal fungus, which was discovered in 1818 and named for its shape like morchella esculenta. The nutrient components of the morchella can be equivalent to cow milk, meat and fish meal. Therefore, it is internationally often referred to as one of "health foods". The morchella esculenta also has the functions of benefiting intestines and stomach, digesting and assisting food, reducing phlegm and regulating qi, tonifying kidney, strengthening yang, tonifying brain and refreshing, and has good treatment effects on weakness of spleen and stomach, dyspepsia, excessive phlegm, short breath, dizziness and insomnia. There is a saying that the lamb tripe is eaten every year and the eighty mountains are walked as usual in folk.
The earliest record about the cultivation of the morchella is 1883 years, and a plurality of morchella cultivation fruiting reports are found over 100 years later, wherein the symbolic breakthrough is the success of the lower artificial control cultivation in 1982, and then the research on the cultivation of the morchella enters a rapid development stage. 24 months in 1994, 8.5.8.5.5.5, Terry farm Inc. announced to be built into the first morchella cultivation factory in the world, 2002, Yulong county, Ludian county, Lihongbin, etc. adopted fresh and dried fruit bodies as strains, and then used round-leaf poplar as an auxiliary material to cultivate morchella; in 2003, Zhaoqi et al used purely cultured morchella conica mycelia to be sown in farmlands and return to forests, and a small amount of round-leaved poplar was added as an auxiliary material, and the success was achieved in 2004. The commercial cultivation of morchella esculenta is realized in 2012 by the research of the product of the year 27.
Although the cultivation of the morchella esculenta has made great progress, many scientific and technical problems are not solved, the industrial development of the morchella esculenta is severely restricted, the cultivation risk is also the point, and the manufacture of the nutrition bag for the cultivation of the morchella esculenta is one of the key problems. At present, the nutrition bag plays a key role in field cultivation and development of morchella, before the cultivation of morchella has not been broken through for many years, the domestic field cultivation technology is realized in a differentiated way by the concept of several patents in the United states 80 years for the Tan Fang and the teacher in Sichuan academy of forestry, the technology treatment is called as hypha touch technology at first, and the Guoyong teacher in Sichuan academy of agriculture at that time changes the vertical bag into the horizontal swing bag and changes the vertical bag into the external aid nutrition bag. In the commercial cultivation stage of the morchella, cultivation without external nutrition is basically failed, and the loss of external nutrition bags under the current technical system has great influence on yield. Scientific experiments also show that the manufacturing process of the nutrition bag also has a crucial influence on the yield.
Disclosure of Invention
The invention aims to provide a compound microbial inoculum for morel cultivation external aid nutrition, aiming at the problems of the existing morel cultivation external aid nutrition.
The invention also aims to provide a nutrient material for the external nutrition of morchella cultivation.
The invention also aims to provide a using method of the nutrient material for the morchella cultivation external aid nutrition.
In order to achieve the purpose, the invention adopts the technical scheme that: a compound microbial preparation for supplementing nutrition for Morchella esculenta cultivation comprises Cellulomonas flavigena, Candida utilis and Geotrichum candidum. Preferably, the feed additive comprises 3-5 parts of cellulomonas flavigena, 2-4 parts of candida utilis and 2-4 parts of geotrichum candidum in parts by weight.
A Morchella esculenta cultivation external-aid nutritional material is prepared from matrix, the above compound microbial inoculum and water; the substrate consists of rice hulls, straws, mushroom bran, gypsum, glucose, salt and potassium dihydrogen phosphate; the addition amount of the composite microbial inoculum is 4-6% of the weight of the matrix; the addition amount of the water is to adjust the water content of the nutrient material to 60-65%.
The substrate comprises, by weight, 30-35 parts of rice hulls and 30-35 parts of straws; 30-35 parts of mushroom bran, 1-2 parts of gypsum, 2-3 parts of glucose, 1-2 parts of salt and 0.1-0.3 part of monopotassium phosphate.
A morchella cultivation external-aid nutrient material is prepared by the following steps:
1) respectively carrying out expanded culture on cellulomonas flavigena, candida utilis and geotrichum candidum in a liquid culture medium; preferably, the liquid culture medium is: 1-20g of soluble starch, 1-10g of yeast extract, 1-5g of peptone, 0.1-3g of potassium nitrate, 0.1-10 mg/L of manganese sulfate, 0.1-10 mg/L of sodium molybdate, 1-6g of dipotassium hydrogen phosphate, 0.8-5 g of sodium chloride, 0.5-4 g of magnesium sulfate and 1000 ml of water, and the pH value is adjusted to 7.2-7.4.
2) Straw pretreatment: crushing the straws to 0.5-1 cm;
3) pretreatment of mushroom bran: drying the fungus bran until the water content is lower than 10%;
4) taking rice hulls, pretreated straws, pretreated mushroom bran, gypsum, glucose, salt and potassium dihydrogen phosphate, uniformly mixing, adding water to adjust the water content to be 60-65%, adding a composite microbial inoculum with the weight of 4-6% of the matrix, uniformly mixing, piling into a round pile, then covering with a film, fermenting, turning once when the material temperature reaches 55-58 ℃, continuing to ferment, and spreading out and cooling when the material temperature reaches 55-58 ℃ again.
A method for using exogenous nutrient material for culturing morel in culturing morel. The method comprises the following steps: filling the morchella cultivation external-aid nutrient material into a plastic fungus bag, pricking a plurality of holes on the same side of the material bag, wherein the hole diameter is 1-2cm, and fastening two ends; 2 lines are sequentially arranged on two sides of the morchella bed, and the opening is tightly covered with soil.
The invention has the beneficial effects that: the complex microbial inoculum of the invention has symbiotic coordination without mutual antagonism, and can provide sufficient external nutrition after being fermented with the substrate. The invention utilizes the compound microbial inoculum and the mushroom bran rich in growth promoting substances as external nutrition, and can obviously improve the yield and accelerate the germination speed of the morchella esculenta. The manufacturing process and the using method have the advantages of simplicity, convenience, practicability, good effect and low cost.
Detailed Description
The active ingredients of the complex microbial inoculum comprise the following strains: cellulomonas flavigena (preservation number ACCC 04313), Candida utilis (preservation number ACCC 20059), Geotrichum candidum (preservation number ACCC 21170), all purchased from China agricultural microbial culture Collection management center.
Preparing a slant culture medium. The specific formula is as follows: 1-20g of soluble starch, 1-10g of yeast extract, 1-5g of peptone, 0.1-3g of potassium nitrate, 0.1-10 mg/L of manganese sulfate, 0.1-10 mg/L of sodium molybdate, 1-6g of dipotassium hydrogen phosphate, 0.8-5 g of sodium chloride, 0.5-4 g of magnesium sulfate, 20g of agar and 1000 ml of water, and the pH value is adjusted to 7.2-7.4. Heating the culture medium to dissolve in water, sterilizing at 121 deg.C for use, sterilizing for 30min, and packaging into culture dish.
Respectively inoculating the strains of cellulomonas flavigena, candida utilis and geotrichum candidum to a slant culture medium, standing and culturing at a constant temperature of 38 ℃ for 24h, and detecting no infectious microbe infection by a microscope, wherein the spore rate is more than 90%. The cultured strain is preserved at a temperature below 3 ℃.
And (4) preparing a liquid culture medium. The specific formula is as follows: 1-20g of soluble starch, 1-10g of yeast extract, 1-5g of peptone, 0.1-3g of potassium nitrate, 0.1-10 mg/L of manganese sulfate, 0.1-10 mg/L of sodium molybdate, 1-6g of dipotassium hydrogen phosphate, 0.8-5 g of sodium chloride, 0.5-4 g of magnesium sulfate and 1000 ml of water, and the pH value is adjusted to 7.2-7.4. Heating the culture medium to dissolve in water, packaging into strain bottles, sterilizing at 121 deg.C for use, and sterilizing for 30 min.
Inoculating the strain cultured in the slant culture medium into a liquid culture medium, and performing constant temperature shaking culture at the conditions of 160-200rpm and 33-35 ℃ for 22-24 h to respectively obtain the liquid strains of cellulomonas flavigena, candida utilis and geotrichum candidum.
Example 1A Morchella esculenta cultivation external-aid nutrient
Composite bacterial preparation
PDA slant culture medium: the formula of the culture medium is as follows: 15g of soluble starch, 5g of yeast extract, 3g of peptone, 1g of potassium nitrate, 5mg/L of manganese sulfate, 6mg/L of sodium molybdate, 6g of dipotassium hydrogen phosphate, 3g of sodium chloride, 4 g of magnesium sulfate, 20g of agar and 1000 ml of water, and the pH value is adjusted to 7.2. Sterilizing at 121 deg.C for 30min, and packaging into culture dish.
Inoculating strains of cellulomonas flavigena, candida utilis and geotrichum candidum to a slant culture medium, standing and culturing at a constant temperature of 38 ℃ for 24 hours, detecting no infectious microbe infection by a microscope, and ensuring that the spore rate is more than 90 percent. The cultured strain is preserved at a temperature below 3 ℃.
Liquid culture medium: the formula of the culture medium is as follows: 15g of soluble starch, 5g of yeast extract, 3g of peptone, 1g of potassium nitrate, 1mg/L of manganese sulfate, 3mg/L of sodium molybdate, 3g of dipotassium hydrogen phosphate, 5g of sodium chloride, 1.5 g of magnesium sulfate and 1000 ml of water, and the pH value is adjusted to 7.4. Subpackaging in strain bottles, sterilizing the culture medium at 121 deg.C for use, and sterilizing for 30 min.
Inoculating the strain cultured in the PDA slant culture medium into a liquid culture medium, and performing constant temperature oscillation culture at 33 ℃ at 180rpm for 24h to respectively obtain the liquid strains of cellulomonas flavigena, candida utilis and geotrichum candidum.
Carrying out liquid strains of cellulomonas flavigena, candida utilis and geotrichum candidum in a weight ratio of 5: 3: 3 to obtain the composite microbial inoculum.
Preparation and application of external-aid nutrient material for morchella cultivation
1) Straw pretreatment: crushing the straws to 0.5-1 cm;
2) pretreatment of mushroom bran: drying the fungus bran until the water content is lower than 10%;
3) the matrix formula is as follows: 35 parts of rice hulls; 30 parts of straw; 30 parts of mushroom bran; 3 parts of edible glucose; 2 parts of gypsum; 1 part of table salt; and 0.1 part of monopotassium phosphate.
4) Mixing rice hull, straw, mushroom bran, edible glucose, gypsum, salt and potassium dihydrogen phosphate, adding water after stirring uniformly, adjusting the water content to 65%, and adding a composite microbial inoculum, wherein the adding amount of the composite microbial inoculum is 5% of the weight of the matrix.
5) Uniformly mixing, piling into a round pile, covering with a layer of plastic film, turning over once when the material temperature reaches 56 ℃, then continuing fermentation, and spreading out and cooling when the material temperature reaches 56 ℃ again to obtain the morchella cultivation external aid nutrient material.
6) The morchella cultivation external nutrient material is filled into a plastic fungus bag with the thickness of 15cm multiplied by 55cm multiplied by 0.04cm, and two ends are tied tightly. 4 round holes with the diameter of 1cm are tied at the same side of the material bag, 2 lines are sequentially placed at two sides of the morchella bed, the hole is downward, and the material bag is tightly attached to the bed surface and covered with soil.
Example 2A Morchella esculenta cultivation external-aid nutrient
Composite bacterial preparation
PDA slant culture medium: the formula of the culture medium is as follows: 8g of soluble starch, 3g of yeast extract, 2g of peptone, 2g of potassium nitrate, 1mg/L of manganese sulfate, 3mg/L of sodium molybdate, 3g of dipotassium hydrogen phosphate, 5g of sodium chloride, 1.5 g of magnesium sulfate, 20g of agar and 1000 ml of water, and the pH value is adjusted to 7.3. The medium was sterilized at 121 ℃ for 30min and then dispensed into petri dishes.
Inoculating strains of cellulomonas flavigena, candida utilis and geotrichum candidum to a slant culture medium, standing and culturing at a constant temperature of 38 ℃ for 24 hours, detecting no infectious microbe infection by a microscope, and ensuring that the spore rate is more than 90 percent. The cultured strain is preserved at a temperature below 3 ℃.
Liquid culture medium: the formula of the culture medium is as follows: 15g of soluble starch, 5g of yeast extract, 3g of peptone, 1g of potassium nitrate, 1mg/L of manganese sulfate, 3mg/L of sodium molybdate, 3g of dipotassium hydrogen phosphate, 5g of sodium chloride, 1.5 g of magnesium sulfate and 1000 ml of water, and the pH value is adjusted to 7.4. Subpackaging in strain bottles, sterilizing the culture medium at 121 deg.C for use, and sterilizing for 30 min.
Inoculating the strain cultured in the PDA slant culture medium into a liquid culture medium, and performing constant temperature oscillation culture at 200rpm and 35 deg.C for 22h to obtain liquid strains of Cellulomonas flavigena, Candida utilis and Geotrichum candidum respectively.
Carrying out liquid strains of cellulomonas flavigena, candida utilis and geotrichum candidum in a weight ratio of 4: 4: 3 to obtain the composite microbial inoculum.
Preparation and application of external-aid nutrient material for morchella cultivation
1) Straw pretreatment: crushing the straws to 0.5-1 cm;
2) pretreatment of mushroom bran: drying the fungus bran until the water content is lower than 10%;
3) the matrix formula is as follows: 32 parts of rice hull, 30 parts of straw, 30 parts of mushroom residue, 2 parts of edible glucose, 1 part of gypsum, 1 part of salt and 0.1 part of monopotassium phosphate.
4) Mixing rice hull, straw, mushroom bran, edible glucose, gypsum, salt and potassium dihydrogen phosphate, adding water after stirring uniformly, adjusting the water content to 65%, and adding a composite microbial inoculum, wherein the adding amount of the composite microbial inoculum is 5% of the weight of the matrix.
5) Uniformly mixing, piling into a round pile, covering with a layer of plastic film, turning over once when the material temperature reaches 56 ℃, then continuing fermentation, and spreading out and cooling when the material temperature reaches 56 ℃ again to obtain the morchella cultivation external aid nutrient material.
6) The morchella cultivation external nutrient material is filled into a plastic fungus bag with the thickness of 75cm multiplied by 50cm multiplied by 0.02cm, and two ends are fastened. 4 round holes with the diameter of 1.5cm are tied at the same side of the material bag, 2 rows are sequentially placed at two sides of the fungus bed, and the hole positions face downwards and tightly cling to the bed surface.
Carrying out Morchella cultivation test on different exogenous nutrients, wherein the production mode, the cultivation conditions and other conditions are consistent, and the formula of the conventional nutrition bag is as follows: 33% of wheat grains, 33% of rice straws, 33% of sawdust and 1% of gypsum, and the test results are shown in table 1.
TABLE 1
| Example 1 | Example 2 | Control group (conventional medium) | |
| Mean time to initial germination | Day 1.5 | Day 2 | Day 4 |
| Contamination ratio (%) | 0.4 | 0.5 | 4.5 |
| Growth of hypha in bag | ++++ | ++++ | + |
| Fruiting time after placing the nutrition bag | Day 15 | Day 17 | Day 29 |
| Yield (kg/m)2) | 7.5 | 7.2 | 5.1 |
"+, + + + +" indicates the growth of morchella hypha
As is apparent from Table 1, the external-aid nutrient material has the advantages that the effect is outstanding compared with the conventional method, the average time of initial germination is shortened by 2-2.5 days, the pollution rate is reduced by 4-4.1%, the fruiting time is shortened by 12-14 days, and the yield is improved by 2.1-2.4kg/m2。
Claims (5)
1. A morchella cultivation external-aid nutrient material is characterized by being prepared from a matrix, a compound microbial inoculum and water; the substrate comprises, by weight, 30-35 parts of rice hulls, 30-35 parts of straws, 30-35 parts of mushroom bran, 1-2 parts of gypsum, 2-3 parts of glucose, 1-2 parts of salt and 0.1-0.3 part of monopotassium phosphate; the composite microbial inoculum comprises 3-5 parts of cellulomonas flavigena, 2-4 parts of candida utilis and 2-4 parts of geotrichum candidum in parts by weight; the addition amount of the composite microbial inoculum is 4-6% of the weight of the matrix; the addition amount of the water is to adjust the water content of the nutrient material to 60-65%.
2. A preparation method of the morchella cultivation external-aid nutrient material is characterized by comprising the following steps:
1) respectively carrying out expanded culture on cellulomonas flavigena, candida utilis and geotrichum candidum in a liquid culture medium;
2) straw pretreatment: crushing the straws to 0.5-1 cm;
3) pretreatment of mushroom bran: drying the fungus bran until the water content is lower than 10%;
4) taking rice hulls, pretreated straws, pretreated mushroom bran, gypsum, glucose, salt and monopotassium phosphate, uniformly mixing, adding water to adjust the water content to be 60-65%, adding a composite microbial inoculum with the weight of 4-6% of the matrix, uniformly mixing, piling into a circular pile, covering with a film, fermenting, turning once when the material temperature reaches 55-58 ℃, continuing to ferment, and spreading and cooling when the material temperature reaches 55-58 ℃ again.
3. The method according to claim 2, wherein the liquid medium is: 1-20g of soluble starch, 1-10g of yeast extract, 1-5g of peptone, 0.1-3g of potassium nitrate, 0.1-10 mg/L of manganese sulfate, 0.1-10 mg/L of sodium molybdate, 1-6g of dipotassium hydrogen phosphate, 0.8-5 g of sodium chloride, 0.5-4 g of magnesium sulfate and 1000 ml of water, and the pH value is adjusted to 7.2-7.4.
4. The use of the morchella esculenta cultivation external-aid nutrient according to claim 1 for cultivating morchella esculenta.
5. Use according to claim 4, characterized in that the method is as follows: filling the morchella cultivation external-aid nutrient material into a plastic fungus bag, pricking a plurality of holes on the same side of the fungus bag, wherein the hole diameter is 1-2cm, and fastening two ends; 2 lines are sequentially arranged on two sides of the morchella bed, and the opening is tightly covered with soil.
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| CN110073893A (en) * | 2019-05-10 | 2019-08-02 | 绵阳市安州区保兴食用菌农业科技有限公司 | A kind of implantation methods of hickory chick |
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