CN112919982A - Compound microbial agent and fertilizer special for eucalyptus - Google Patents
Compound microbial agent and fertilizer special for eucalyptus Download PDFInfo
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- CN112919982A CN112919982A CN202110211054.2A CN202110211054A CN112919982A CN 112919982 A CN112919982 A CN 112919982A CN 202110211054 A CN202110211054 A CN 202110211054A CN 112919982 A CN112919982 A CN 112919982A
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F3/00—Fertilisers from human or animal excrements, e.g. manure
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G5/00—Fertilisers characterised by their form
- C05G5/10—Solid or semi-solid fertilisers, e.g. powders
- C05G5/12—Granules or flakes
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Abstract
The invention provides a compound microbial agent and a fertilizer special for eucalyptus, and belongs to the technical field of biological fertilizers. The compound microbial agent comprises bacillus megatherium, paenibacillus kribbensis and paenibacillus mucilaginosus, wherein the preservation number of the paenibacillus kribbensis is CGMCC No.17248, and the preservation number of the paenibacillus mucilaginosus is CGMCC No. 3995. The compound microbial fertilizer comprises a compound microbial agent, an organic fertilizer and a compound inorganic fertilizer. The compound microbial agent and the fertilizer can improve the utilization rate of the fertilizer, improve the quality of soil and crops and accelerate the yield of eucalyptus.
Description
Technical Field
The invention relates to the technical field of biological fertilizers, in particular to a compound microbial agent special for eucalyptus and a fertilizer.
Background
Eucalyptus is one of the most various, fastest growing and most widely used species of trees in the world, is the most widely cultivated broad-leaved tree species in the world at present, more than 100 countries have been introduced and developed in a large scale in the world, the area of the eucalyptus breaks through 2000 million hectares, and the eucalyptus becomes an important resource concerned in the world. The wood provided every year accounts for 37% of the global artificial forest wood yield, and plays an important role in relieving the contradiction between supply and demand of wood, guaranteeing the global wood safety and coping with global climate change.
At present, a large number of artificial cultivated species of eucalyptus are used, the variety of the used fertilizer is more, the quick-acting fertilizer is mainly used, and the nutrient components required by the rapid growth of the eucalyptus are met. For example, patent CN1718565A provides a eucalyptus fertilizer, which is used for fertilizing eucalyptus artificial forests. The special compound fertilizer containing N, P, K elements and Mn, Mo, Zn, Cu, Fe, B and other elements is provided according to the growth and nutritional requirements of eucalyptus, so that the high growth of eucalyptus can be increased by 20% on average, and the yield of single plant can be increased by 25% on average. Patent CN103553781B discloses a eucalyptus fertilizer additive, which comprises 3-6 parts of humic acid; 3-10 parts of trace elements; 3-9 parts of rare earth elements; 2-4 parts of a slow release agent; 3-6 parts of a water-retaining agent; 6-12 parts of organic matters can meet the requirement of the growth cycle of eucalyptus on a large amount of various nutrient elements, improve the utilization rate of the fertilizer and improve the volume of eucalyptus. Patent CN105198589A discloses a eucalyptus controlled-release fertilizer, which comprises the following raw materials in parts by weight: 60-160 parts of a wrapping material and 1400 parts of fertilizer particles, wherein the eucalyptus controlled-release fertilizer is prepared by the steps of crushing, extracting, filtering, concentrating, fermenting, granulating, drying, cooling, wrapping and the like. The fertilizer can effectively improve the utilization efficiency of the fertilizer, promote the eucalyptus to grow fast and healthily and improve the growth speed of the eucalyptus.
The application of the slow release material can prolong the using time of the fertilizer, reduce the fertilizing times and reduce the loss rate of the fertilizer. However, as the afforestation area of eucalyptus is continuously enlarged, the land capability is reduced, the composition, structure and function of soil microbial communities are changed, the ecological system is degraded in multiple functions, the yield of eucalyptus is reduced, the soil fertility is reduced, and the subsequent planting requirements are influenced. The multifunctional microbial fertilizer is applied to fundamentally solve the problem of soil biological community restoration and the technical problem of increasing the growth speed of eucalyptus which is urgently needed to be solved.
Disclosure of Invention
In view of the above, the invention aims to provide the compound microbial agent and the fertilizer special for eucalyptus, which can promote the rhizosphere development of crops, provide all-round nutrition for eucalyptus, improve the disease resistance of eucalyptus, and have certain quality-improving and yield-increasing effects on eucalyptus.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a compound microbial agent special for eucalyptus, which comprises bacillus megatherium, paenibacillus kribbensis and paenibacillus mucilaginosus, wherein the paenibacillus kribbensis is bacillus kribbensis of 021 type, and the preservation number is CGMCC No. 17248; the paenibacillus mucilaginosus is paenibacillus mucilaginosus No. 01, and the preservation number is CGMCC No. 3995.
The composite microbial agent comprises fermentation liquor of bacillus megatherium, bacillus kribbensis and bacillus mucilaginosus, the total number of effective live bacteria of the bacillus megatherium fermentation liquor is 200-300 hundred million/ml, the number of effective live bacteria of the bacillus kribbensis fermentation liquor is 0.5-2 hundred million/ml, and the number of effective live bacteria of the bacillus mucilaginosus fermentation liquor is 2.0-5.0 hundred million/ml.
Further preferably, the volume ratio of the bacillus megaterium fermentation liquid to the bacillus kribbensis fermentation liquid to the paenibacillus mucilaginosus fermentation liquid is 6-10: 2-4: 3-5.
Preferably, the composite microbial powder is obtained by performing centrifugal dehydration and spray drying on fermentation liquor of bacillus megatherium, bacillus kribbensis and bacillus mucilaginosus.
The invention also provides a compound microbial fertilizer special for eucalyptus, which comprises, by mass, 10-20% of any one of the compound microbial agents, 50-70% of an organic fertilizer and 20-30% of an inorganic compound fertilizer.
According to a preferable technical scheme, the organic fertilizer is obtained by fermenting agricultural wastes through a microbial agent, wherein the agricultural wastes comprise the following components in parts by weight: 35-60 parts of feces, 35-60 parts of mushroom residues, 15-30 parts of one or more organic wastes of straws, wheat bran, bran coat, traditional Chinese medicine residues, seaweed mud and tea residues, wherein the addition amount of the microbial agent is 2-10 parts by weight.
Preferably, the fermentation temperature is 55-65 ℃, and the fermentation time is 15-20 days.
Further preferably, the fermentation also comprises an after-ripening stage of 15-20 d.
Preferably, the inorganic compound fertilizer is nitrogen-phosphorus-potassium fertilizer, and the mass ratio of nitrogen, phosphorus and potassium is 10-14: 4-6: 6-10.
The invention also comprises the application of the compound microbial agent or the compound microbial fertilizer in promoting the growth of eucalyptus.
The invention has the beneficial effects that:
the strain contained in the compound microbial agent special for eucalyptus can generate a plurality of plant endogenous hormones such as indoleacetic acid and the like in the fermentation culture process, promotes the cell division and the elongation of eucalyptus, strengthens metabolism, accelerates the root development of eucalyptus, improves the water and nutrient absorption capacity of eucalyptus, enhances the stress resistance capacity of eucalyptus and improves the growth rate of eucalyptus.
The compound microbial fertilizer special for eucalyptus contains functional microorganisms, nitrogen, phosphorus and potassium and organic matters, has excellent biocontrol characteristics, has a synergistic effect and promotes each other. The compound microbial agent disclosed by the invention can be mixed with a chemical fertilizer to improve the utilization rate of the chemical fertilizer and improve the quality of soil and crops. After the fertilizer is applied, the soil can be loosened, the soil hardening caused by applying the fertilizer can be improved, the formation of a soil granular structure is accelerated, the good air permeability of the soil is ensured, and the eucalyptus lumber production is accelerated.
Detailed Description
The invention provides a compound microbial agent special for eucalyptus, which comprises bacillus megatherium, paenibacillus kribbensis and paenibacillus mucilaginosus, wherein the paenibacillus kribbensis is bacillus kribbensis of 021 type, and the preservation number is CGMCC No. 17248; the paenibacillus mucilaginosus is paenibacillus mucilaginosus No. 01, and the preservation number is CGMCC No. 3995.
The bacillus megaterium is a phosphate-solubilizing bacterium and has a good effect of degrading organic phosphorus in soil. The source of Bacillus megaterium in the present invention is not particularly limited, and Bacillus megaterium species known in the art can be used. In the specific embodiment of the invention, the bacillus megaterium is a commercial product.
The Paenibacillus mucilaginosus No. 01 has the capacity of activating potassium and silicon in silicate minerals and is called as silicate bacteria (potassium bacteria); the bacterium has the following effects:
(1) the fertilizer can play roles of fixing nitrogen, dissolving phosphorus and potassium and releasing soluble phosphorus and potassium elements and medium and trace elements such as calcium, sulfur, magnesium, iron, zinc, molybdenum, manganese and the like during the propagation and growth in soil, thereby not only improving the soil fertility, but also providing absorbable and utilizable comprehensive nutrient elements for crops and preventing and improving the physiological nutrient deficiency lesions of the crops.
(2) The stress resistance and growth promotion function is as follows: the fertilizer also generates various physiological active substances such as gibberellin, indoleacetic acid, cytokinin and the like and protein amino acid substances in the soil metabolism process, can increase the chlorophyll content of crops in a same ratio, obviously enhances the photosynthesis of the crops, promotes the root systems of the crops to be developed and grow healthily, enhances the cold resistance and drought resistance of the crops, secretes chemical substances for inhibiting the growth of plant pathogenic bacteria to the rhizosphere, improves the disease resistance of the crops, improves the yield of the crops and improves the product quality.
(3) Space and nutrient competition: the bacterium has root tropism, occupies ecological sites, isolates pathogenic bacteria, and has iron competitive advantage. The paenibacillus mucilaginosus preservation strain of the invention produces siderophin, quickly exhausts the trace available iron in soil, so that pathogenic bacteria lack iron and cannot propagate, and iron nutrition is provided for plants through the iron-siderophin, so that the plants benefit.
The bacillus like bacillus kribbensis, the three-torch 021 variety, has control effects on various plant diseases, such as sclerotinia rot of colza, rice sheath blight, alternaria alternata and the like; the secondary metabolite polyfructose generated in the fermentation process of the preserved strain also has the growth promoting property, and meanwhile, the strain also has the phosphate-solubilizing effect.
Optionally, the three microorganisms in the microbial agent of the invention are microbial agents obtained by respectively fermenting strains of functional bacteria to prepare fermentation broth, and mixing the obtained fermentation broths according to a certain proportion to obtain a certain proportion of viable bacteria.
As an alternative embodiment, the fermentation method of Bacillus megaterium of the present invention comprises the following steps:
1) activating bacillus megatherium strains, and preparing a seed solution;
2) and inoculating the activated bacillus megaterium prepared into seed solution into a liquid culture medium in a fermentation tank for liquid fermentation, and stopping fermentation when the total number of live bacteria reaches 200-300 hundred million/ml to obtain the bacillus megaterium fermentation solution.
The activation of the bacillus megaterium strain is to inoculate the preserved bacillus megaterium strain into an eggplant-shaped bottle filled with a Chaudhur culture medium for activation culture, wherein the activation culture condition is preferably culture at 28-35 ℃ for 24-36 h, and more preferably culture at 30-33 ℃ for 28-34 h.
The liquid culture medium inoculated by the liquid fermentation of the bacillus megaterium comprises the bacillus megaterium liquid culture medium which is conventionally used in the field, and the liquid culture medium is preferably used after being sterilized for 30-45min at 120-130 ℃. The fermentation equipment is not particularly limited, and the conventional fermentation tank in the technical field can be adopted.
In the invention, the temperature of the liquid fermentation of the bacillus megaterium is preferably 28-35 ℃, and more preferably 30-32 ℃; the pH value of the fermentation is preferably 5.8-6.5, more preferably 6.0; the pressure in the tank is preferably 0.05-0.1 MPa, and more preferably 0.07-0.08 MpPa; the ventilation volume of the liquid fermentation is preferably 0.5-0.8V/V.min, and more preferably 0.6V/V.min; the time for the liquid fermentation is preferably 36 to 48 hours, more preferably 40 hours.
The method preferably detects the total number of the live fermentation bacteria every 3-4 hours after the bacillus megaterium is fermented for 36-40 hours. Preferably, the fermentation is stopped when the total number of the live bacteria reaches 200-300 hundred million/ml, and more preferably, the fermentation is stopped when the total number of the live bacteria reaches 250-299 hundred million/ml, so that the bacillus megaterium fermentation liquid is obtained.
As an alternative embodiment, the fermentation of Paenibacillus kribbensis according to the present invention comprises the activation of the species, shake flask fermentation and fermentor fermentation. Activating the preserved Paenibacillus krikoides with a beef extract peptone culture medium, selecting a single colony or lawn, inoculating the single colony or lawn into a shake flask filled with an improved liquid Cnahs culture medium, and carrying out shake culture at 28-32 ℃ and 180-200 r/min for 36-48h to enable the Paenibacillus krikoides to grow rapidly. When the strain grows to meet the production conditions, namely no sundry bacteria is detected under the microscopic examination, the strain is inoculated into a liquid fermentation tank for liquid fermentation culture, and the raw materials used for fermentation are proportioned as follows:
| media composition | g/L | Secondary seed tank usage |
| Sucrose | 25-30 | 9-10kg |
| Sodium nitrate | 1.0-2.0 | 450-500g |
| Dipotassium hydrogen phosphate | 0.5-1.5 | 300-350g |
| Magnesium sulfate | 0.3-0.6 | 150-200g |
| Potassium chloride | 0.5-1.0 | 150-200g |
| Ferric sulfate | 0.01 | 3-5g |
| Defoaming agent | 900-1000ml |
And (4) culturing for 4-5 days according to the fermentation amount of 50% after inoculation. And in the fermentation process, controlling the temperature at 28-30 ℃, continuously introducing oxygen, continuously stirring, and taking out of the tank when the bacillus Paenibacillus Clarkii fermentation liquor is in a viscous state to obtain the bacillus Paenibacillus Clarkii fermentation liquor. The effective viable count of the fermentation liquor is 0.5-2 hundred million/ml, preferably 2 hundred million/ml.
As an alternative embodiment, the fermentation method of paenibacillus mucilaginosus provided by the invention comprises the following steps:
1) activating the strain of the Paenibacillus mucilaginosus, and preparing a liquid;
2) inoculating the activated paenibacillus jelly sample into a liquid culture medium in a fermentation tank for liquid fermentation to obtain the paenibacillus jelly sample fermentation liquid.
The activation of the paenibacillus mucilaginosus strain in the invention is to inoculate the preserved paenibacillus mucilaginosus strain in an eggplant-shaped bottle filled with a Chao's culture medium, and culture the strain at 28-32 ℃ for 36-48h for activation culture.
The composition of the liquid culture medium inoculated by the liquid fermentation of the paenibacillus mucilaginosus in the invention is the paenibacillus mucilaginosus liquid culture medium which is conventionally used in the field, and the raw material formula used for the fermentation is as follows:
| media composition | Percentage of | Secondary seed tank usage | Preference is given to |
| Starch | 0.03％ | 90-100g | 100g |
| Yeast powder | 0.03％ | 90-110g | 100g |
| Soybean meal powder | 0.025％ | 75-85g | 80g |
| Magnesium sulfate | 0.007％ | 21-27g | 24g |
| Dipotassium hydrogen phosphate | 0.003％ | 9-11g | 10g |
| Calcium carbonate | 0.003％ | 9-11g | 10g |
| Sodium chloride | 0.0005％ | 1.5-2.5g | 2g |
| Defoaming agent | 3％ | 900-1000g | 950g |
The liquid culture medium is preferably used after being sterilized for 30-45min at 120-130 ℃. The fermentation equipment is not particularly limited, and the conventional fermentation tank in the technical field can be adopted.
In the invention, the temperature of the liquid fermentation of the Paenibacillus mucilaginosus is preferably 28-32 ℃, and more preferably 30 ℃; the pH value of the liquid fermentation is preferably 5.8-6.5, more preferably 6.0; the pressure in the tank is preferably 0.05-0.1 MPa, and more preferably 0.08 MpPa; the ventilation volume of the liquid fermentation is preferably 0.5-0.8V/V.min, and more preferably 0.6V/V.min; the time for the liquid fermentation is preferably 48 to 60 hours, more preferably 52 hours.
In the invention, the stirring mode during fermentation of the Paenibacillus mucilaginosus is preferably that the fermentation tank is not opened for the first 12 hours, is opened once an hour for 12-24 hours for about 10 seconds each time, is opened once a half hour for about 10 seconds each time after 24 hours, is 10 minutes/time and is 10 seconds once after the fermentation tank is taken out for several hours before the fermentation tank is taken out, and is stirred all the time. The microscopic examination is carried out every 4 hours after 32 hours, the fermentation liquor can not be taken out of the tank (standard position for tank taking) after 56 hours of observation, and if the fermentation liquor can not be taken out of the tank for continuous culture, the effective viable count of the fermentation liquor of the Paenibacillus mucilaginosus produced by the method is 2.0-5.0 hundred million/ml, and preferably 5.0 hundred million/ml.
In the microbial agent, the volume ratio of the bacillus megaterium fermentation liquid to the bacillus kribbensis fermentation liquid to the bacillus mucilaginosus fermentation liquid is 6-10: 2-4: 3 to 5, and more preferably 8:3: 4.
The compound microbial agent can also be microbial powder. And respectively carrying out centrifugal dehydration and spray drying on the fermentation liquor of the bacillus megatherium, the bacillus kribbensis and the bacillus mucilaginosus to obtain fermentation bacteria powder of each microorganism, and then mixing the bacteria powder according to a proportion to obtain the compound microorganism bacteria powder. Or mixing the fermentation liquor of the bacillus megatherium, the bacillus kribbensis and the bacillus mucilaginosus in proportion, and then carrying out centrifugal dehydration and spray drying to obtain the composite microbial powder.
The invention also provides a compound microbial fertilizer special for eucalyptus, which comprises, by mass, 10-20% of the compound microbial agent, 50-70% of an organic fertilizer and 20-30% of an inorganic compound fertilizer.
In the invention, the organic fertilizer is obtained by fermenting agricultural wastes through microorganisms, wherein the agricultural wastes comprise the following components in parts by weight: 35-60 parts of feces, 35-60 parts of mushroom residues, 15-30 parts of one or more organic wastes of straws, wheat bran, bran coat, traditional Chinese medicine residues, seaweed mud and tea residues, wherein the addition amount of the microbial fermentation liquid is 2-10 parts by weight. Preferably, the agricultural waste comprises the following components in parts by weight: 40-50 parts of feces, 40-50 parts of mushroom residues, 20-25 parts of one or more organic wastes of straws, wheat bran, bran coat, traditional Chinese medicine residues, seaweed mud and tea residues, wherein the microorganism is preferably fermentation liquor of the microorganism, and the addition amount is preferably 3-5 parts by weight. The invention has no special limitation on the types of organic fertilizer fermentation microorganisms, and the conventional organic fertilizer fermentation microorganisms in the field can be adopted. The fermentation of the organic fertilizer adopts oxygen-introducing fermentation, and preferably adopts an oxygen-exposing fermentation tank for oxygen-introducing fermentation. Controlling the temperature in the fermentation process at 55-65 ℃, preferably 60-62 ℃; keeping aeration and turning over the plate, and fermenting at a temperature of not more than 75 ℃. Keeping recording the changes of water content, material pile height, material pile porosity and color in the fermentation; the fermentation period in the whole tank is controlled to be 15-20 days, and the after-ripening time after the tank is taken out is controlled to be 15-20 days. Sieving, crushing, sterilizing and drying the after-ripening materials to prepare the organic fertilizer.
In the invention, the inorganic compound fertilizer is preferably a nitrogen-phosphorus-potassium fertilizer, and the mass ratio of nitrogen, phosphorus and potassium is 10-14: 4-6: 6-10, more preferably 12:5: 8.
the compound microbial agent, the organic fertilizer and the inorganic compound fertilizer are mixed according to a proportion and granulated to obtain the granular compound microbial fertilizer.
The specific method for granulating is not particularly limited, and the conventional granulating method in the field is adopted, for example, the mixed semi-finished product is sent into an extrusion type granulator for granulating, so that the compound microbial fertilizer special for eucalyptus is obtained.
The compound microbial agent and the fertilizer special for eucalyptus contain functional microorganisms, nitrogen, phosphorus and potassium and organic matters, have excellent biocontrol characteristics, have synergistic effects with each other, promote each other, can effectively improve the growth speed of eucalyptus and improve the height and the breast diameter of eucalyptus.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Preparing functional bacteria fermentation liquor:
preparing a bacillus megaterium fermentation liquid: the preserved bacillus megaterium strain is inoculated into an eggplant-shaped bottle filled with a Chaudhur culture medium and activated and cultured for 24 hours at 32 ℃. Inoculating the activated bacillus megatherium into a fermentation tank for liquid fermentation, wherein the temperature of the liquid fermentation is 32 ℃, and the pH value of the liquid fermentation is 6.0; the pressure in the tank was 0.07MPa, and the ventilation was 0.6V/V.min. And detecting the total number of the fermented live bacteria every 3-4 hours after fermenting for 36-40 hours, and stopping fermenting when the total number of the live bacteria reaches 300 hundred million/ml to obtain the bacillus megaterium fermentation liquor.
Preparing a Paenibacillus fermentation liquid of Krebs: activating bacillus Paenibacillus Clarke with the preservation number of CGMCC No.17248 with a beef extract peptone culture medium, selecting a single colony or lawn, inoculating the single colony or lawn into a shake flask containing an improved liquid Cnahs culture medium, and carrying out shake culture at 30 ℃ and 200r/min for 48h to ensure that the bacillus Paenibacillus Clarke grows rapidly. When no foreign bacteria are detected in microscopic examination, the culture medium is inoculated into a liquid fermentation tank for liquid fermentation culture, and the raw materials used for fermentation are proportioned as follows: 30g/L of sucrose, 2.0g/L of sodium nitrate, 1.0g/L of dipotassium phosphate, 0.5g/L of magnesium sulfate, 0.8g/L of potassium chloride and 0.01g/L of ferric sulfate. And in the fermentation process, controlling the temperature at 30 ℃, continuously introducing oxygen, continuously stirring, and taking out the fermentation liquor of the Paenibacillus kribbensis from the tank when the fermentation liquor of the Paenibacillus kribbensis is in a viscous state to obtain the fermentation liquor of the Paenibacillus kribbensis, wherein the effective viable count of the fermentation liquor is 2 hundred million/ml.
Preparing a paenibacillus mucilaginosus fermentation liquid: inoculating the Bacillus mucilaginosus strain with the preservation number of CGMCC No.3995 into an eggplant-shaped bottle filled with a Chaudhur culture medium, and performing activation culture at 30 ℃ for 48 h. The activated Paenibacillus mucilaginosus is fermented in the following liquid culture media: 0.03% of starch, 0.03% of yeast powder, 0.025% of soybean meal, 0.007% of magnesium sulfate, 0.003% of dipotassium hydrogen phosphate, 0.003% of calcium carbonate, 0.0005% of sodium chloride, 3% of defoaming agent and water, and the pH value of the fermentation liquor is kept to be 6.0. The temperature of liquid fermentation is 30 ℃, the pressure in the tank is 0.08MPa, the ventilation volume is 0.6V/V.min, the time of liquid fermentation is 52 hours, and the effective viable count of the fermentation liquid of the gel-like paenibacillus is 5.0 hundred million/ml.
Example 2
Compound microbial agent
The paenibacillus kribbensis fermentation liquor, the paenibacillus mucilaginosus fermentation liquor and the arthrobacter nicotianae fermentation liquor in the embodiment 1 are uniformly mixed according to the volume ratio of 8:3:4 to obtain the compound microbial agent.
Example 3
Uniformly mixing the paenibacillus kribbensis fermentation liquor, the paenibacillus mucilaginosus fermentation liquor and the arthrobacter nicotianae fermentation liquor in the volume ratio of 6:2:5, and carrying out centrifugal dehydration and spray drying on the mixed fermentation liquor to obtain the compound microbial agent.
Example 4
And (3) respectively carrying out centrifugal dehydration on the paenibacillus kribbensis fermentation liquor, the paenibacillus mucilaginosus fermentation liquor and the arthrobacter nicotianae fermentation liquor in the volume ratio of 10:4:3, carrying out spray drying, and mixing to obtain the microbial powder to obtain the compound microbial agent.
Example 5
Composite microbial fertilizer
According to the following steps of 35: 50: 15, weighing the excrement, the mushroom dregs and the seaweed mud, and crushing to obtain 100-mesh particles. Adding the crushed raw materials into an organic material decomposing microbial inoculum, conveying the mixture to an oxygen aeration fermentation tank for oxygen aeration fermentation, and controlling the temperature in the fermentation process to be 65 ℃; keeping aeration and turning over the plate, and fermenting at a temperature of not more than 75 ℃. Recording the changes of water content, material pile height, material pile porosity and color in the fermentation; the fermentation period in the whole tank is controlled at 15d, and the after-ripening time after discharging the tank is controlled at 15 d. Sieving, crushing, sterilizing and drying the after-ripening materials to prepare the organic fertilizer.
By weight percentage, 10% of the compound microbial agent of the embodiment 2, 70% of the organic fertilizer and 20% of the inorganic compound fertilizer are mixed, and are granulated by an extrusion type granulator to obtain the compound microbial fertilizer. Wherein the mass ratio of nitrogen, phosphorus and potassium in the organic compound fertilizer is 12:5: 8.
Example 6
According to the following steps of 45: 35: 10: 10, weighing the excrement, the mushroom residue, the seaweed mud and the straws in a weight ratio, and crushing to obtain 100-mesh particles. Adding the crushed raw materials into an organic material decomposing microbial inoculum, conveying the mixture to an oxygen aeration fermentation tank for oxygen aeration fermentation, and controlling the temperature in the fermentation process to be 55 ℃; keeping aeration and turning over the plate, and fermenting at a temperature of not more than 75 ℃. Recording the changes of water content, material pile height, material pile porosity and color in the fermentation; the fermentation period in the whole tank is controlled at 20d, and the maturation after discharging is controlled at 20 d. Sieving, crushing, sterilizing and drying the after-ripening materials to prepare the organic fertilizer.
And (2) mixing 20% of the compound microbial agent in the embodiment 2, 50% of the organic fertilizer and 30% of the inorganic compound fertilizer in percentage by weight, and granulating by using an extrusion type granulator to obtain the compound microbial fertilizer. Wherein the mass ratio of nitrogen, phosphorus and potassium in the organic compound fertilizer is 10:6: 9.
Example 7
Effect of compound microbial fertilizer on growth of eucalyptus
1. The test eucalyptus variety is Eucalyptus grandis with the age of 2.5 years.
Test soil: the soil of the test field is red and red soil, the soil layer of 0-60cm has 12.1g/kg of organic matters, 0.57g/kg of total nitrogen, 10.9mg/kg of alkaline hydrolysis nitrogen, 3.8mg/kg of available phosphorus, 10.8mg/kg of quick-acting potassium and the pH value of 4.8. The soil has low content of available nutrients and low fertility level.
And (3) experimental design: the experiment was performed in a random block array with 4 treatments and 3 repetitions, 40 plants per cell (242.4 m area)2) Experimentally at a rate of 666.7m2And planting 110 eucalyptus plants.
Treatment 1: example 5 composite microbial fertilizer + 80% conventional fertilization;
and (3) treatment 2: sterilized example 5 composite microbial fertilizer + 80% conventional fertilization;
and (3) treatment: conventional fertilization;
and (4) treatment: blank control.
And (4) fertilization and management record: two ditches are respectively arranged between each eucalyptus plant, the fertilizing ditch is 60cm long, 20cm wide and 20-30 cm deep, and fertilizer is evenly applied into the ditches and then covered with soil. Example 5 composite microbial fertilizer, sterilized example 5 composite microbial fertilizer per 666.7m2The using amount is 24kg, which is converted into 8.7kg of compound microbial fertilizer or compound microbial fertilizer without live bacteria applied to each cell, and 0.22kg of compound microbial fertilizer is applied to each eucalyptus. The normal fertilizer is 666.7m in each2Special compound fertilizer (N: P) for applying 35% eucalyptus2O5∶K2O is 18:7:10)55kg, calculated by conversion20kg of eucalyptus oil is applied to each cell, and 0.5kg of eucalyptus oil is applied to each cell. 80% of conventional fertilizer is used for every 666.7m244kg of 35% compound fertilizer special for eucalyptus is applied, and the application amount is converted into 16.0kg of compound fertilizer for each cell and 0.4kg of compound fertilizer for each eucalyptus.
The test was carried out in 2019, 3 months and 18 days, and the tree vigor was investigated once each 3 months, 6 months, 9 months and 12 months after fertilization before fertilization. And collecting soil samples during the last investigation, partially air-drying, and storing part of fresh soil in a refrigerator at 4 ℃ to be tested.
The tree vigor investigation method comprises the following steps: 6 eucalyptus trees with similar growth vigor are selected for tracking and monitoring in each cell, and the tree height and the diameter at breast height are measured by using a check ruler of each tree. And performing data statistical analysis by using an EXCEL table.
2. Results
(1) Effect of different treatments on the Tree height of Eucalyptus at different times
As can be seen from table 1, the eucalyptus height was significantly increased after the investigation in 2019, 6/18 th, 2019, 9/18 th, 2019, 12/18 th and 2020, 3/18 th compared to before the fertilization. The tree heights of 3 months, 6 months, 9 months and 12 months after the treatment of 1 fertilization are respectively increased by 0.96m, 1.23m, 2.50m and 3.16m than those before the fertilization, and the tree heights of 2 months, 3 months and 4 months after the treatment of 1 fertilization are respectively increased by 7.0%, 8.9% and 53.3%, and the tree heights are obviously increased; the height of the trees after the treatment 2 and the fertilization of 3 months, 6 months, 9 months and 12 months is respectively increased by 0.85m, 0.94m, 2.09m and 2.73m compared with the height of the trees before the fertilization of 2 months, and the height of the trees after the treatment 2 and the fertilization of 12 months is respectively increased by 1.7 percent and 43.3 percent compared with the height of the trees after the treatment 2 and the fertilization of 4 months; the tree heights of 3 months, 6 months, 9 months and 12 months after the treatment of 3 fertilizers are respectively increased by 0.63m, 0.90m, 2.10m and 2.61m compared with the tree heights before the fertilization, and the tree heights of 12 months after the treatment of 3 fertilizers are increased by 40.9% compared with the tree heights of 4 months after the treatment of 3 fertilizers, so that the tree heights are obviously increased.
TABLE 1 Effect of different treatments on Eucalyptus Tree height
Remarking: the data in the table are the average values of the cells.
(2) Analysis of variance of tree height after fertilization of eucalyptus with different treatments for 12 months
The tree height condition table (shown in table 2) shows that after the eucalyptus trees are fertilized for 12 months in different treatment modes, the average tree height of the eucalyptus trees in the treatment mode 1 is 5.81m, which is increased by 0.38m and 7.0% compared with the treatment mode 2, and is increased by 0.47m and 8.9% compared with the treatment mode 3; an increase of 2.02m over treatment 4, an increase of 53.3%.
TABLE 2 Table of the height of the tree after applying fertilizer to eucalyptus for 12 months
(3) Influence of different treatments on the diameter of the breast at different times of eucalyptus
As can be seen from Table 3, the chest diameters of eucalyptus trees in different time periods after fertilization are obviously increased compared with those before fertilization. After the treatment of 1, the chest diameter ratios of 3 months, 6 months, 9 months and 12 months after fertilization are respectively increased by 1.63cm, 2.82cm, 2.95cm and 3.39cm before fertilization, and after the treatment of 1, the chest diameter ratios of 12 months after fertilization are respectively increased by 6.1%, 8.9% and 49.0% for the treatments of 2, 3 and 4; after the treatment 2, the chest diameter ratios of 3 months, 6 months, 9 months and 12 months after fertilization are respectively increased by 1.53cm, 2.54cm, 2.89cm and 3.12cm before fertilization, and after the treatment 2, the chest diameter ratios of 12 months after fertilization are respectively increased by 2.6 percent and 40.4 percent after treatment 3 and 4; the chest diameter ratios of 3 months, 6 months, 9 months and 12 months after the treatment of 3 fertilizers are respectively increased by 1.37cm, 2.46cm, 2.71cm and 2.97cm before the fertilizers are applied, and the chest diameter ratios of 12 months after the treatment of 3 fertilizers are increased by 36.8 percent and are obviously increased.
TABLE 3 Effect of different treatments on the diameter at breast height of Eucalyptus
Remarking: the data in the table are the average values of the cells.
(4) Analysis of variance of breast diameter after fertilization of eucalyptus with different treatments for 12 months
The chest diameter condition table (shown in table 4) shows that after the eucalyptus is fertilized for 12 months in different treatments, the average chest diameter of the treatment 1 is 5.30cm, which is increased by 0.30cm and 6.1% compared with the treatment 2; the increase is 0.43cm and 8.9 percent compared with the treatment 3; an increase of 1.74cm, 49.0% over treatment 4.
TABLE 4 chest diameter after fertilization of eucalyptus for 12 months with different treatments
To summarize: after the eucalyptus is applied with the compound microbial fertilizer and 80% of conventional fertilization for 12 months, the difference of the tree height is obvious and increased by 70% compared with the tree height obtained by the non-viable bacterial fertilizer and 80% of conventional fertilization, and the breast diameter is greatly increased by 6.1%; compared with the conventional fertilization treatment, the tree height and the breast diameter are obviously increased by 8.9 percent and 8.9 percent respectively.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. The compound microbial agent special for eucalyptus is characterized by comprising bacillus megatherium, paenibacillus kribbensis and paenibacillus mucilaginosus, wherein the paenibacillus kribbensis is bacillus kribbensis No. 021, and the preservation number is CGMCC No. 17248; the paenibacillus mucilaginosus is paenibacillus mucilaginosus No. 01, and the preservation number is CGMCC No. 3995.
2. The compound microbial agent as claimed in claim 1, wherein the compound microbial agent comprises fermentation liquor of bacillus megaterium, bacillus kribbensis and bacillus mucilaginosus, the total effective viable bacteria of the bacillus megaterium fermentation liquor is 200-300 hundred million/ml, the effective viable bacteria number of the bacillus kribbensis fermentation liquor is 0.5-2 hundred million/ml, and the effective viable bacteria number of the bacillus mucilaginosus fermentation liquor is 2.0-5.0 hundred million/ml.
3. The compound microbial inoculant according to claim 2, wherein the volume ratio of the bacillus megaterium fermentation broth to the bacillus kribbensis fermentation broth to the bacillus mucilaginosus fermentation broth is 6-10: 2-4: 3-5.
4. The compound microbial agent according to any one of claims 1 to 3, wherein fermentation broth of Bacillus megaterium, Bacillus kribbensis and Bacillus mucilaginosus is subjected to centrifugal dehydration and spray drying to obtain the compound microbial powder.
5. A compound microbial fertilizer special for eucalyptus is characterized by comprising, by mass, 10-20% of the compound microbial agent as claimed in any one of claims 1-4, 50-70% of an organic fertilizer, and 20-30% of an inorganic compound fertilizer.
6. The compound microbial fertilizer as claimed in claim 5, wherein the organic fertilizer is obtained by fermenting agricultural wastes with a microbial agent, and the agricultural wastes comprise the following components in parts by weight: 35-60 parts of feces, 35-60 parts of mushroom residues, 15-30 parts of one or more organic wastes of straws, wheat bran, bran coat, traditional Chinese medicine residues, seaweed mud and tea residues, wherein the addition amount of the microbial agent is 2-10 parts by weight.
7. The compound microbial fertilizer as claimed in claim 6, wherein the fermentation temperature is 55-65 ℃ and the fermentation time is 15-20 d.
8. The composite microbial fertilizer according to claim 6, further comprising an after-ripening stage of 15-20d after fermentation.
9. The compound microbial fertilizer as claimed in claim 5, wherein the inorganic compound fertilizer is nitrogen, phosphorus and potassium fertilizer, and the mass ratio of nitrogen, phosphorus and potassium is 10-14: 4-6: 6-10.
10. The use of the compound microbial inoculant of any one of claims 1 to 4 or the compound microbial fertilizer of any one of claims 5 to 9 for promoting the growth of eucalyptus.
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