CN106116883A - A kind of preparation method of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer - Google Patents

Abstract

The invention relates to a preparation method of compound microbial bacterial fertilizer for root irrigation of peony, comprising the following steps: fermenting sweet potato dregs and fruit wastes by Candida tropicalis to obtain fermented product I; mixing chicken manure, cow dung, biogas tank The slag and wheat straw were mixed and naturally fermented to obtain the fermented product II; the fermented product I was mixed with the fermented product II in an appropriate proportion, and fermented with Geotrichum candidum, Bacillus megaterium and Bacillus jelly-like to obtain the fermented product III; Add an appropriate amount of mycorrhizal fungal spores of Glomus intrarhizogi and Glomus moesei, mycelia inside and outside the root, and the mixture of infected plant root segments, as well as fish meal, zinc sulfate, and Bacillus subtilis HS5B5 bacterial liquid, and mix well That is, the compound microbial bacterial fertilizer for root irrigation of peony is obtained. According to the growth nutrition and environmental requirements of peony, the bacterial fertilizer prepared by using agricultural by-products as raw materials can provide protection for peony roots, inhibit pathogenic microorganisms, provide comprehensive nutrition for peony growth, promote growth, and prolong flowering period.

Description

A kind of preparation method of compound microbial bacterial fertilizer for root irrigation of peony

technical field

The invention relates to the field of preparation methods of microbial bacterial fertilizer, in particular to a preparation method of compound microbial bacterial fertilizer for peony root irrigation.

Background technique

Peony is a famous ornamental flower in my country. Its flowers are huge, graceful and luxurious, and are deeply loved by people. Peony grows in neutral or slightly alkaline soil, and it is not resistant to acid and heavy soil. Peony likes fertilizers, especially fertilizers with relatively complete elements such as nitrogen, phosphorus, and potassium. At present, the commonly used fertilizers are inorganic compound fertilizers or organic fertilizers such as livestock and poultry manure. Peony is a fleshy root, and the application of inorganic compound fertilizer must be applied far away from the main root. If the fertilization is too close to the main root, it will easily cause water loss in the peony root, which will affect the absorption of mineral elements. Long-term use of inorganic compound fertilizer will easily cause soil damage. Compaction and pollution of groundwater resources; the application of organic fertilizer must be retting and decomposing, and the undecomposed organic fertilizer will ferment and release organic acids, which will cause the root of peony to rot. In addition, during the growth process of peony, it is vulnerable to the influence of adverse environmental conditions and the infection of pathogenic microorganisms. The leaves, branches and rhizomes of peony can become the parts that are harmful to pathogenic microorganisms. The current solution is usually to apply chemical pesticides or Cutting off diseased branches and leaves, etc., but long-term and large-scale application of chemical pesticides can easily cause pathogenic microorganisms to develop resistance, and at the same time cause environmental pollution and ecological balance damage. As peonies are ornamental flowers, the application of chemical pesticides will also threaten the health of tourists.

With the development of agriculture, there are more and more by-products and wastes. They pile up and ferment in large quantities, causing serious pollution to the environment. The rancid smell and harmful substances released have a negative impact on people's lives. It is also a good solution to produce bio-fertilizers.

The Bacillus subtilis (Bacillus subtilis) HS5B5 in the present invention was preserved in the General Microorganism Center (CGMCC) of China Microbiological Culture Collection Management Committee (CGMCC) on May 9, 2012, address: No. 3, Courtyard No. 1, Beichen West Road, Chaoyang District, Beijing, China Institute of Microbiology, Academy of Sciences, deposit number: CGMCC No.6088 (see the description of the invention patent application with application number 201410163640.4). Bacillus subtilis HS5B5 has inhibitory effect on tree peony leaf spot with Cladosporium peony as the pathogenic fungus through biological antagonism, and also has certain inhibitory effect on other common fungal diseases.

Contents of the invention

The object of the present invention is to provide a kind of preparation method of compound microbial bacterial fertilizer for root irrigation of peony, the compound microbial bacterial fertilizer prepared by this method can not only provide various nutrients for the growth of peony, but also can inhibit the growth of pathogenic microorganisms of peony, for Peony roots provide microbial protection.

In order to solve the above-mentioned technical problems, the technical solution adopted in the present invention is: a kind of preparation method of compound microbial bacterial fertilizer for root irrigation of peony, comprising the following steps:

Step 1: Activate Candida tropicalis, cultivate it to the logarithmic growth phase, and obtain mixture A containing Candida tropicalis; take sweet potato residue and fruit waste at a mass ratio of 1:4, crush them to below 1.5 cm, and use sweet potato Based on the total mass of slag and fruit waste, add 1% urea and stir evenly to obtain mixture B; inoculate mixture A into mixture B, the inoculum amount is 1% of the mass of mixture B, stir and ferment at room temperature for 4-5 days, and prepare Obtain fermented product I;

Step 2: Take chicken manure, cow dung, biogas residue and wheat straw crushed to less than 1 cm according to the mass ratio of 1:1:2-3:2-3, and mix them evenly to obtain mixture C. Based on the quality of mixture C, Add 5% quicklime, expose to the air, and let it stand naturally for 3-5 days to obtain the fermented product II;

Step 3. Add the fermented product II prepared in step 2 to the fermented product I prepared in step 1, wherein the mass ratio of fermented product II to fermented product I is 3:1, and mix well to obtain a mixture D; in the mixture D Inoculate Geotrichum candidum, Bacillus megaterium and Bacillus jelly-like in the logarithmic growth phase, and the inoculum amount of each strain is 1% of the mass of the mixture D, stir and ferment at 30°C for 4-6 days to obtain the fermented Object III;

Step 4, take alfalfa as the host plant, propagate Glomus intrarhizoi in the propagation matrix, and the inoculum size is 3%-5%, when the Glomus intrarhizoi spores reach more than 30 per gram of soil, inoculate Broken alfalfa roots and propagation substrates to obtain a mixture E1 containing the mycorrhizal fungus spores of Glomus intrarhizoi, hyphae inside and outside the roots and infected root segments; alfalfa is used as the host plant to propagate Glomus moses in the propagation substrate, The inoculum amount is 3%-5%. When the spores of Glomus mosesi reach more than 30 per gram of soil, smash the alfalfa root and the propagation substrate to obtain the mycorrhizal fungus spores containing Glomus mosesi mycorrhizae, hyphae inside and outside the root and The mixture E2 that infects the root segment; mix the mixture E1 and the mixture E2 in a mass ratio of 1:1 to obtain the mixture E;

Step 5. Take the fermented product III, mixture E, fish meal and zinc sulfate according to the mass ratio of 100:90:2:1, add the mixture E, fish meal and zinc sulfate to the fermented product III, and mix well to obtain a semi-solid mixture F;

Step 6, taking Bacillus subtilis HS5B5, activating it in the seed medium, and transferring it to fermentation medium after activation to ferment, and when the Bacillus subtilis HS5B5 grows to the stationary phase, a mixture G containing Bacillus subtilis HS5B5 is obtained;

Step 7. Mix the mixture F prepared in step 5 with the mixture G prepared in step 6 at a mass ratio of 3-5:1, and stir evenly to obtain a compound microbial fertilizer for root irrigation of peony.

Further, the fruit waste described in step 1 is any one or any mixture of fallen apples, rotted apples, fresh apple pomace, and pineapple pomace.

Further, the fruit waste in step 1 is a mixture of fresh apple residue and fallen apple fruit, and the mass ratio of fresh apple residue and fallen apple fruit is 1:3.

Further, the sweet potato residue and fruit waste described in step one are crushed to below 0.8cm.

Further, the mass ratio of chicken manure, cow dung, biogas residue and wheat straw in step 2 is 1:1:2:3.

Further, in step 4, the propagation substrate for the propagation of Glomus intrarhizitis and Glomus mosesi is prepared by mixing river sand and soil at a mass ratio of 1:1 and then sterilizing.

Further, the seed medium described in step six is a nutrient broth medium.

Further, the activation temperature of the Bacillus subtilis HS5B5 described in step 6 is 30° C., and the activation time is 11 hours.

Further, the fermentation medium described in step 6 is based on adding 1000ml of water as a reference, and its components and additions are respectively: corn flour 15.0g, glucose 3.0g, soybean flour 15.0g, peptone 3.0g, MgSO ₄ . 7H ₂ O 0.5g, KH ₂ PO ₄ ·2H ₂ O 1.0g, water 1000ml, adjust the pH to 7.0.

Further, the fermentation condition of the Bacillus subtilis HS5B5 described in step 6 in the fermentation medium is: 30° C. for aerated fermentation for 25 hours.

Beneficial effect:

Under the treatment of various raw materials and microorganisms in the present invention, the various components in the prepared compound microbial bacterial fertilizer for tree peony root irrigation can cooperate with each other, and finally achieve the goal of promoting the growth of peony, prolonging the flowering period, and preventing and controlling peony diseases. Purpose;

The compound microbial bacterial fertilizer for root irrigation of peony prepared by the present invention enters the soil through root irrigation, and a delicate dynamic balance is formed between various microorganisms in the bacterial fertilizer and soil microorganisms, so that various organic substances in the bacterial fertilizer are transformed between microorganisms And use, and finally release the nutrients that peony can directly absorb and use into the soil, and then be absorbed and utilized by peony. During the growth and reproduction of Glomus intrarhizoi and Glomus mosesi in the soil, most of the hyphae spread around the rhizosphere of peony and symbiosis with the root system of peony, which increases the absorption area of nutrient substances by root system of peony, which is beneficial to the growth of phosphorus in peony. At the same time, Glomus intraradices, Glomus moses and Bacillus subtilis HS5B5 cooperate with each other to resist the invasion of pathogenic microorganisms on tree peony; the addition of zinc sulfate can significantly improve the stress resistance and multipotency of tree peony. Improve the resistance of various diseases, accelerate the formation of chlorophyll, luxuriant branches and leaves, and promote the vegetative growth of peony;

The raw materials used in the present invention are sweet potato dregs, fresh apple dregs, fallen apples, chicken manure, cow dung, biogas tank slag, and wheat straw, all of which are cheap and easy-to-obtain agricultural by-products, waste or household garbage. After being treated by the method provided by the invention , not only can alleviate the pressure of environmental pollution, but also the prepared bacterial fertilizer can meet the needs of peony for nutrients, and at the same time, it has a good effect on improving soil permeability and enhancing soil fertility, and can also promote the growth of peony root system and prolong the flowering period. Inhibits the growth of peony pathogenic microorganisms and provides microbial protection for peony roots.

detailed description

The present invention will be described in further detail below in conjunction with specific examples.

A preparation method for compound microbial bacterial fertilizer for root irrigation of peony, comprising the following steps:

Step 1. Activate Candida tropicalis in wort medium (add 130 g of malt extract powder and 0.1 g of chloramphenicol for every 1000 ml of water; adjust the pH to 6.0), and cultivate it to the logarithmic growth phase to obtain mixture A, mixture A contains wort medium and rapidly growing Candida tropicalis and its metabolites; the growth of Candida tropicalis with logarithmic growth is inserted into the new medium, which can make Candida tropicalis adapt faster The new growth environment shortens the lag period, thereby shortening the fermentation time;

According to the mass ratio of 1:4, sweet potato residue and fruit waste are taken, crushed to less than 1.5cm, based on the total mass of sweet potato residue and fruit waste, 1% urea is added, and the mixture is evenly stirred to obtain a mixture B; the mixture B is essentially A medium prepared from sweet potato dregs and fruit wastes that can be used to propagate Candida tropicalis, not only solves the problem of agricultural wastes (i.e. sweet potato dregs and fruit wastes) accumulating corruption and polluting the environment, but also alleviates the In addition, sweet potato dregs and fruit wastes also provide available carbon sources, nitrogen sources, inorganic salts, water and other nutrients for the growth and reproduction of Candida tropicalis, adding 1% urea to maximize the carbon-nitrogen ratio In good condition, it is conducive to the growth and reproduction of Candida tropicalis; sweet potato dregs and fruit wastes are crushed to less than 1.5cm. During stirring and fermentation, Candida tropicalis can fully contact with mixture B, which is beneficial to the growth of Candida tropicalis ;

Inoculate mixture A into mixture B, the inoculum amount is 1% of the mass of mixture B, stir and ferment at room temperature for 4-5 days to obtain fermentation product I; , Candida tropicalis decomposes most of the macromolecular nutrients in sweet potato residue and fruit waste into small molecular compounds that are easy to use, and synthesizes a large amount of bacterial protein during the growth process. After the growth cycle of Candida tropicalis, its The bacterial protein can be utilized by subsequent Geotrichum candidum, Bacillus megaterium, Bacillus jelly-like and soil microorganisms; Candida tropicalis will produce a large amount of water and carbon dioxide during the growth and reproduction process, as well as a large amount of undecomposed Organic acids, so the fermented product I obtained is a slightly acidic semi-solid mixture;

Step 2. Take chicken manure, cow dung, biogas digester residue and wheat straw crushed to less than 1cm according to the mass ratio of 1:1:2-3:2-3, and mix them evenly to obtain mixture C, chicken manure, cow dung and biogas digester The slag can be used as organic fertilizer for plants, which itself contains a large amount of organic matter. If the three are used separately, it will cause a large amount of nutrient loss, low utilization rate, and contain a large number of pathogenic spores and parasite eggs, which has a strong odor. Improper burning of seedlings is also easy to cause; chicken manure, cow dung and biogas residues contain a large number of different types of microorganisms, including aerobic microorganisms, anaerobic microorganisms and facultative anaerobic microorganisms, chicken manure, cow dung and biogas digesters The mixture of slag and slag makes the microbial species and metabolites in the mixture C more abundant, which is conducive to the transformation, utilization and release of nutrients among various microorganisms into the soil, and is beneficial to the absorption and utilization of various nutrients by peony; Adding an appropriate proportion of wheat straw crushed to less than 1cm in mixture C can maintain a certain degree of air permeability in mixture C, make the soil loose and breathable, meet the growth needs of peony fleshy roots, and at the same time meet the needs of aerobic microorganisms and facultative anaerobes Microbes’ demand for oxygen; based on the quality of mixture C, add 5% quicklime, expose to the air, and let it stand naturally for 3-5 days to obtain fermentation product II. When quicklime encounters chicken manure, cow manure and biogas pit residue The water in the mixture C will get alkaline slaked lime, and with a large amount of heat to form high temperature, the high temperature and the alkalinity of slaked lime can not only kill most of the pathogenic bacteria and parasite eggs in the mixture C, but also make the wheat straw further decompose, segregate and After 3-5 days of natural fermentation, most of the pathogenic bacteria and parasite eggs in the mixture C are killed, and the mixing of various raw materials is also more uniform, and the macromolecular substances in the raw materials are transformed into more Nutrients that can be directly absorbed and utilized by peony;

Step 3. Add the fermented product II prepared in step 2 to the fermented product I prepared in step 1. The alkalinity of the fermented product II can be neutralized with the acidity of the fermented product I. The mass ratio of the fermented product II to the fermented product I is The pH value of the mixture D obtained after mixing is 3:1, so that the pH value of the mixture D is suitable for the subsequent microbial fermentation, and the compound microbial bacterial fertilizer for peony roots obtained finally does not damage the fleshy roots of peony; the inoculation in the mixture D is in logarithmic growth candidum, Bacillus megaterium and Bacillus jelly-like, and the inoculum amount of each strain is 1% of the mass of mixture D, stirred and fermented at 30°C for 4-6 days to obtain ferment III; After the initial separation and degradation of wheat straw, macromolecular substances such as cellulose and lignin in it are further decomposed by Geotrichum candidum and Bacillus megaterium to produce dextrin, oligosaccharides and monosaccharides, which can be used by fermentation strains and soil microorganisms ; jelly-like bacillus can release potassium elements in insoluble potassium salts and other medium and trace elements during the growth and reproduction process, which is beneficial to the absorption and utilization of these elements by peony, and has a positive effect on prolonging the flowering period of peony. jelly-like bacillus The organic acids and other substances in the metabolites of the peony can be directly absorbed and utilized by the peony to promote the growth of the peony;

Step 4, take alfalfa as the host plant, multiply Glomus intrarhizoi in the propagation matrix, and the inoculum size is 3%-5%, when the Glomus intrarhiza spores reach more than 30 per gram of soil, the At that time, mycorrhizal fungi had invaded the root system of plants in large quantities, smashed alfalfa roots and propagation substrates, and obtained mixture E1 containing spores of mycorrhizal fungi of Glomus intraradices, hyphae inside and outside roots, and infected root segments; alfalfa was used as the host plant , multiply Glomus moesei in the propagation medium, the inoculum amount is 3%-5%, when the spores of Glomus moesei reach more than 30 per gram of soil, the mycorrhizal fungus has already infected the plants in large quantities For the root system, crush the alfalfa root and the propagation substrate to obtain a mixture E2 containing the mycorrhizal fungus spores of Glomus mosesii, hyphae inside and outside the root and infected root segments; mix the mixture E1 and the mixture E2 uniformly in a mass ratio of 1:1 to obtain the mixture E. During the growth and reproduction of Glomus intrarhizoi and Glomus mosei in the soil, most of the hyphae spread around the rhizosphere of peony and symbiosis with the root system of peony, which increases the absorption area of root system of peony, which is beneficial to the root system of peony. Absorption of mineral elements such as phosphorus, and at the same time, Glomus intrarhiza, Glomus moses and Bacillus subtilis HS5B5 cooperate with each other to jointly resist the invasion of pathogenic microorganisms in the soil to peony;

Step 5. Take the fermented product III, mixture E, fish meal and zinc sulfate according to the mass ratio of 100:90:2:1, add the mixture E, fish meal and zinc sulfate to the fermented product III, and mix well to obtain a semi-solid Mixture F: The protein content in fishmeal is relatively high, and a large amount of small peptides and amino acids are produced after degradation. After absorption by peony, it can enhance the metabolic function of peony, improve photosynthesis, prolong the flowering period, promote the growth of peony root system, and improve the soil micro-ecological environment; sulfuric acid Zinc can significantly improve the stress resistance of peony itself and the ability to resist various diseases, accelerate the formation of chlorophyll, luxuriant branches and leaves, and promote the vegetative growth of peony;

Step 6: Get Bacillus subtilis HS5B5, activate it in the seed culture medium, transfer it to the fermentation medium after activation to ferment, when the Bacillus subtilis HS5B5 grows to the stationary phase, obtain the mixture G containing Bacillus subtilis HS5B5, and ferment at this time Nutrients in the medium were exhausted, and mixture G mainly contained a large amount of bacteria (>10 ⁸ CFU/ml), spores and secondary metabolites; Bacillus subtilis HS5B5 had significant antagonism against pathogenic microorganisms of various diseases of tree peony Bacteriostasis, can cooperate with Glomus intraroote and Glomus mosesi to enhance the disease resistance of peony roots; at the same time, in the process of using the final compound microbial fertilizer, also spray Bacillus subtilis on the aboveground part of peony Bacillus HS5B5 bacterial agent, after this treatment, it will form an all-round protection for the roots, stems and leaves of peony, and the disease resistance effect is more obvious than that of simply using bacterial fertilizer to irrigate the roots;

Step 7. Mix the mixture F prepared in step 5 with the mixture G prepared in step 6 at a mass ratio of 3-5:1, and stir evenly to obtain a compound microbial fertilizer for root irrigation of peony.

Further, the fruit waste described in step 1 is any one or any mixture of fallen apples, rotten apples, fresh apple residues, and pineapple residues, all of which contain A large amount of acidic organic matter, its acidity and beneficial microbial flora can inhibit the reproduction of pathogenic microorganisms in peony, improve the degree of salinization of the land, and significantly increase fertility.

Further, the fruit waste described in step 1 is a mixture of fresh apple residue and fallen apple fruit, the mass ratio of fresh apple residue and fallen apple fruit is 1:3; pectin and protein in sweet potato residue, fresh apple residue and fallen apple fruit And cellulose can be degraded by microorganisms in the bacterial fertilizer to obtain substances such as amino acids, polypeptides and fatty acids, which are conducive to the absorption of peony, promote the growth of peony, and improve the stress resistance of peony.

Further, the sweet potato residue and fruit waste described in the first step are crushed to less than 0.8 cm, which is beneficial to shorten the fermentation time.

Further, the mass ratio of chicken manure, cow dung, biogas residue and wheat straw in step 2 is 1:1:2:3.

Further, in step 4, the propagation substrate for the propagation of Glomus intrarhizitis and Glomus mosesi is prepared by mixing river sand and soil at a mass ratio of 1:1 and then sterilizing.

Further, the seed medium described in step 6 is a nutrient broth medium; the fermentation medium described in step 6 takes every 1000ml of water added as a reference, and its components and additions are respectively: corn flour 15.0g, glucose 3.0g, soybean powder 15.0g, peptone 3.0g, MgSO ₄ 7H ₂ O 0.5g, KH ₂ PO ₄ 2H ₂ O 1.0g, water 1000ml, adjust the pH to 7.0; the activation temperature of Bacillus subtilis HS5B5 The temperature is 30°C, and the activation time is 11 hours; the conditions for the fermentation of Bacillus subtilis HS5B5 in the fermentation medium described in step 6 are: 30°C for aerated fermentation for 25 hours.

The compound microbial bacterial fertilizer for tree peony root irrigating prepared by the method provided by the invention sprays the tree peony stems and leaves with the Bacillus subtilis HS5B5 bacterial agent while irrigating the peony roots, which can make the peony plants strong, enhance disease resistance, and improve the peony flowers. quality, prolong the flowering period, and increase the ornamental effect.

Example:

Candida tropicalis, Geotrichum candidum, Bacillus megaterium, Bacillus jelly-like, Glomus intraradices and Glomus mosei used in the following examples were all purchased from Shandong Shenhai Biotechnology Co., Ltd. The preservation numbers are respectively: Candida tropicalis is preserved in China General Microorganism Culture Collection Management Center, and the preservation number is CGMCC 2.2735. Geotrichum candidum is preserved in the China General Microorganism Culture Collection and Management Center, and the preservation number is CGMCC 2.1132. Bacillus megaterium (Bacillus megaterium) is preserved in the China General Microorganism Culture Collection and Management Center, and the preservation number is CGMCC 1.2416. The jelly-like Paenibacillus (Bacillus mucilaginosus) is preserved in the China Agricultural Microorganism Culture Collection Management Center, and the preservation number is ACCC 10012. Glomus intraradices and Glomus mosseae are registered and preserved by the European Culture Collection, and the preservation numbers are BEG 141 and BEG 167, respectively.

Example 1

A preparation method for compound microbial bacterial fertilizer for root irrigation of peony, comprising the following steps:

Step 1: Take Candida tropicalis and activate it in the wort medium (add 130 g of malt extract powder and 0.1 g of chloramphenicol for every 1000 ml of water; adjust the pH to 6.0), cultivate it to the logarithmic growth phase, and obtain Candida tropicalis-containing Yeast mixture A; according to the mass ratio of 1:1:3, take sweet potato dregs, fresh apple dregs and fallen apples, crush them to less than 0.8cm, and add 1% urea based on the total mass of sweet potato dregs, fresh apple dregs and fallen apples , stir evenly to obtain mixture B; inoculate mixture A into mixture B, the inoculum amount is 1% of the mass of mixture B, stir and ferment at room temperature for 5 days, and obtain fermented product I;

Step 2. Take chicken manure, cow dung, biogas residue and wheat straw crushed to less than 1cm according to the mass ratio of 1:1:2:3, and mix them evenly to obtain mixture C. Based on the quality of mixture C, add 5% of quicklime, exposed to the air and placed naturally for 5 days to obtain fermented product II;

Step 3. Add the fermented product II prepared in step 2 to the fermented product I prepared in step 1, wherein the mass ratio of fermented product II to fermented product I is 3:1, and mix well to obtain a mixture D; in the mixture D Inoculate Geotrichum candidum, Bacillus megaterium and Bacillus jelly-like in the logarithmic growth phase, and the inoculum amount of each strain is 1% of the mass of the mixture D, stir and ferment at 30°C for 6 days to obtain the fermentation product III ;

Step 4, take alfalfa as the host plant, propagate Glomus intrarhizoi in the propagation matrix, the inoculum size is 5%, when the Glomus intrarhiza spores reach more than 30 in every gram of soil, smash the root of alfalfa and propagation substrate, to obtain the mixture E1 containing the mycorrhizal fungus spores of Glomus intraradices, mycelium inside and outside the root, and the infected root segment; alfalfa is used as the host plant, and Glomus mosei is propagated in the propagation substrate, and the inoculum size is 5%, when the spores of Glomus mosesi reach more than 30 per gram of soil, smash the alfalfa root and propagation substrate to obtain a mixture containing the mycorrhizal fungus spores of Glomus mosesi, hyphae inside and outside the root and infected root segments E2; mix mixture E1 and mixture E2 uniformly at a mass ratio of 1:1 to obtain mixture E; the propagation substrates used for the propagation of Glomus intrarhizoi and Glomus moesei are made of river sand and soil at a mass ratio of 1:1 Prepared by mixing and sterilizing;

Step 5. Take the fermented product III, mixture E, fish meal and zinc sulfate according to the mass ratio of 100:90:2:1, add the mixture E, fish meal and zinc sulfate to the fermented product III, and mix well to obtain a semi-solid mixture F;

Step 6: Take Bacillus subtilis HS5B5, activate it in the seed medium at 30°C for 11 hours, then transfer it to the fermentation medium at 30°C for aeration and fermentation for 25 hours, so that the Bacillus subtilis HS5B5 grows to the stationary phase, and obtains Bacillus subtilis containing Mixture G of HS5B5;

The described seed culture medium is a nutrient broth culture medium, with every 1000ml of water added as a reference, its components and additions are respectively: beef extract 5.0g, peptone 10.0g, NaCl 5.0g, water 1000ml, adjust the pH to 7.0;

The fermentation medium is based on the addition of 1000ml of water, and its components and amounts are: corn flour 15.0g, glucose 3.0g, soybean flour 15.0g, peptone 3.0g, MgSO ₄ ·7H ₂ O 0.5g , KH ₂ PO ₄ ·2H ₂ O 1.0g, water 1000ml, adjust the pH to 7.0;

Step 7. Mix the mixture F prepared in step 5 with the mixture G prepared in step 6 at a mass ratio of 5:1, and stir evenly to obtain a compound microbial fertilizer for root irrigation of peony.

Example 2

A preparation method for compound microbial bacterial fertilizer for root irrigation of peony, comprising the following steps:

Step 1: Take Candida tropicalis and activate it in the wort medium (add 130 g of malt extract powder and 0.1 g of chloramphenicol for every 1000 ml of water; adjust the pH to 6.0), cultivate it to the logarithmic growth phase, and obtain Candida tropicalis-containing Yeast mixture A; according to the mass ratio of 1:1:3, take sweet potato dregs, fresh apple dregs and fallen apples, crush them to less than 0.8cm, and add 1% urea based on the total mass of sweet potato dregs, fresh apple dregs and fallen apples , stir evenly to obtain mixture B; inoculate mixture A into mixture B, the inoculum amount is 1% of the mass of mixture B, stir and ferment at room temperature for 4 days, and obtain fermented product I;

Step 2. Take chicken manure, cow dung, biogas residue and wheat straw crushed to less than 1cm according to the mass ratio of 1:1:2:3, and mix them evenly to obtain mixture C. Based on the quality of mixture C, add 5% of quicklime, exposed to the air and placed naturally for 3 days to obtain fermented product II;

Step 3. Add the fermented product II prepared in step 2 to the fermented product I prepared in step 1, wherein the mass ratio of fermented product II to fermented product I is 3:1, and mix well to obtain a mixture D; in the mixture D Inoculate Geotrichum candidum, Bacillus megaterium and Bacillus jelly-like in the logarithmic growth phase, and the inoculation amount of each strain is 1% of the mass of the mixture D, stir and ferment at 30°C for 4 days to obtain the fermentation product III ;

Step 4, take alfalfa as the host plant, multiply Glomus intraradices in the propagation matrix, and the inoculum size is 3%, and when the Glomus intraradices spores reach more than 30 per gram of soil, smash the roots of alfalfa and propagation substrate, to obtain the mixture E1 containing the mycorrhizal fungus spores of Glomus intraradices, mycelium inside and outside the root, and the infected root segment; alfalfa is used as the host plant, and Glomus mosei is propagated in the propagation substrate, and the inoculum size is 3%, when the spores of Glomus moseae reach more than 30 per gram of soil, smash the alfalfa root and propagation substrate to obtain a mixture containing the mycorrhizal fungus spores of Glomus moseae, hyphae inside and outside the root and infected root segments E2; mix mixture E1 and mixture E2 uniformly at a mass ratio of 1:1 to obtain mixture E; the propagation substrates used for the propagation of Glomus intrarhizoi and Glomus moesei are made of river sand and soil at a mass ratio of 1:1 Prepared by mixing and sterilizing;

Step 5. Take the fermented product III, mixture E, fish meal and zinc sulfate according to the mass ratio of 100:90:2:1, add the mixture E, fish meal and zinc sulfate to the fermented product III, and mix well to obtain a semi-solid mixture F;

Step 6: Take Bacillus subtilis HS5B5, activate it in the seed medium for 11 hours at 30°C, and then transfer it to the fermentation medium: 30°C for aeration and fermentation for 25 hours, so that the Bacillus subtilis HS5B5 grows to the stable phase, and obtains subtilis containing Mixture G of Bacillus HS5B5;

The described seed culture medium is a nutrient broth culture medium, with every 1000ml of water added as a reference, its components and additions are respectively: beef extract 5.0g, peptone 10.0g, NaCl 5.0g, water 1000ml, adjust the pH to 7.0;

The fermentation medium is based on the addition of 1000ml of water, and its components and amounts are: corn flour 15.0g, glucose 3.0g, soybean flour 15.0g, peptone 3.0g, MgSO ₄ ·7H ₂ O 0.5g , KH ₂ PO ₄ ·2H ₂ O 1.0g, water 1000ml, adjust the pH to 7.0;

Step 7. Mix the mixture F prepared in step 5 with the mixture G prepared in step 6 at a mass ratio of 3:1, and stir evenly to obtain a compound microbial fertilizer for root irrigation of peony.

test:

The compound microbial bacterial fertilizer for root irrigation of peony (hereinafter referred to as special bacterial fertilizer for peony) prepared by the method provided in Example 1 and the intermediate product obtained in the preparation process were tested.

Experiment 1. One month before the peony flowering period, select 400 peonies with the same growth in the peony cultivation area of Luoyang International Peony Garden, and randomly divide them into eight groups ①, ②, ③, ④, ⑤, ⑥, ⑦ and ⑧, with 50 plants in each group , each group is processed as follows:

① Irrigate the roots with special bacterial fertilizer for peony, and spray the branches and leaves of peony with Bacillus subtilis HS5B5 bacterial agent at the same time;

② Irrigate the roots with special bacterial fertilizer for peony, and spray the branches and leaves of peony with water at the same time;

③Irrigate the roots with fermented product III, and spray the peony branches and leaves with water at the same time;

④ Irrigate the roots with the mixture D, and spray the peony branches and leaves with water at the same time;

⑤ Irrigate the roots with water, and spray the branches and leaves of peony with Bacillus subtilis HS5B5 bacterial agent simultaneously;

⑥ Irrigate the roots with water, and spray the peony branches and leaves with water at the same time;

⑦Irrigate the roots with ordinary microbial fertilizer, and spray the peony branches and leaves with the Bacillus subtilis HS5B5 bacterial agent at the same time;

⑧ Irrigate the roots with ordinary microbial fertilizer, and spray the peony branches and leaves with water at the same time;

In the above treatment, the peony special bacterial fertilizer, fermented product III, mixture D and common microbial bacterial fertilizer used for root irrigation are all used at 80g/plant, and diluted with water to 1000ml for root irrigation; root;

The Bacillus subtilis HS5B5 inoculum used for spraying tree peony branches and leaves in the above-mentioned treatment is the mixture G in the preparation process of Example 1, used by 10g/plant, diluted with water to 500ml and evenly sprayed branches and leaves; ②, ③, ④, ⑥ and ⑧, Use 500ml of water to spray branches and leaves of each peony.

Repeat processing once every 15 days, observe and record florescence, new branch length, flowering rate, flower diameter, single flower quality and infection fungal disease plant number after 30 days (the fungal diseases mainly refer to common tree peony fungal diseases such as peony brown spot disease, below Described fungal disease is identical with this), the result is as shown in table 1:

The impact of various treatments on the flowering of peony in table 1

In Table 1, peony is an ornamental flower, the longer the flowering period of the whole peony, the higher the income; the new branch length represents the growth of the flowering branch in the current year, the longer the branch and the upright flowers, the situation of hidden flowers in the leaves is obtained. Improvement can increase the flower viewing effect; flower formation rate reflects the nutrient development of flower buds; flower diameter and single flower quality both reflect the flower size, and the larger the flower, the better the viewing effect; a, b, c and d all represent different data The significance of the difference, the difference between the same letter is not significant, but the difference between different letters is significant.

The results showed that: group ① had an average entire flowering period of 8.21 days, with the longest new shoots and huge corolla. The flowering rate of 100% indicated that all new shoots in the upper part of the plant could form flowers, indicating that the nutritional status of the plants was good, and there was no abortion of flower buds. At the same time, only one tree peony was infected with fungal diseases; the difference between group ② and group ① was not obvious, but 8 tree peonies were infected with fungal diseases, indicating that root irrigation with special bacterial fertilizer for peony can protect the roots of peony, and to a certain extent, prevent fungal diseases of peony If the tree peony branches and leaves are sprayed with Bacillus subtilis HS5B5 bacterial agent at the same time, it can form all-round protection for tree peony branches, leaves, roots and stems, and can better inhibit fungal pathogens, and the control effect on tree peony fungal diseases is also better; group ③ compared with group ②, the flowering period is shortened by nearly 1 day, the flowering rate, flower diameter and single flower quality are significantly lower than group ①, and the number of infected plants is also more, indicating that the arbuscular branches in mixture E The mycorrhizal mixture, fishmeal, and zinc sulfate can increase and enrich the absorption of nutrients, promote photosynthesis, prolong the flowering period, improve the stress resistance of peony, and reduce the occurrence of diseases; the ④ group uses the mixture D, and its indicators are better than the ③ group Slightly worse, analysis shows that mixture D contains a certain amount of nitrogen, carbohydrates, protein and a small amount of biochemical fulvic acid that are not easily absorbed and utilized by peony; ⑤ group uses Bacillus subtilis HS5B5 bacterial agent (i.e. mixture G) to spray branches and leaves , can prevent the occurrence of peony fungal diseases to a certain extent, but at the same time, only water is used to irrigate the roots. Compared with the group ①, the indicators are significantly lower, and the peony lacks a lot of nutrients, which reduces the stress resistance of the plants. ① Compared with ⑦ or ② and ⑧, the average flowering period is extended by more than 20%, and the flowering rate and flower quality are also greatly increased, which shows that the special bacterial fertilizer for peony is better than ordinary microbial bacterial fertilizer for improving the quality of peony and prolonging the flowering period of peony . Based on the above analysis, the application of the special bacterial fertilizer for peony prepared in Example 1 can improve the stress resistance and flower quality of peony, prolong the flowering period, promote the flowering of peony, and increase the number of flowering. Mixture G, namely Bacillus subtilis HS5B5 inoculum, can also significantly reduce the probability of tree peonies being infected with fungal diseases after spraying branches and leaves. Because of its liquid state characteristics, it can be used in combination with spraying and root irrigation in practical applications.

Experiment 2. During the vegetative growth stage of peonies, 400 peonies with the same growth potential in the peony cultivation area of Luoyang International Peony Garden were selected and randomly divided into eight groups. The specific grouping and treatment of each group were the same as those in Experiment 1.

Repeat processing once every 15 days, record the peony vegetative growth situation after 45 days, the result is as shown in table 2:

Table 2 Effects of various treatments on the vegetative growth of peony

In Table 2, SPAD represents leaf greenness, which reflects the chlorophyll content of the leaves, and together with the net photosynthetic rate, reflects the photosynthetic ability of the plant; increasing the stomatal conductance can reduce stomatal restrictions and promote the increase of photosynthesis, but it cannot Avoided increased transpiration and water loss; the increase of water use efficiency indicates that the increase of stomatal conductance will enhance transpiration, but the effect of promoting photosynthesis is more obvious; a, b, c and d all indicate the significance of the difference between different data, The difference between the same letter is not significant, but the difference between different letters is significant.

The results showed that: group ① had the best growth of tree peonies, and only 2 tree peonies were infected with fungal diseases; the difference between group ② and group ① was not obvious, but 7 tree peonies were infected with fungal diseases. Spraying tree peony branches with Bacillus subtilis HS5B5 can better antagonize fungal pathogens and have a better control effect on tree peony fungal diseases; compared with group ②, the photosynthetic rate is significantly lower, indicating the lack of arbuscular mycorrhiza and fish meal , zinc sulfate, not only reduces the absorption area of the rhizosphere, but also lacks amino acids and zinc, resulting in a decrease in photosynthesis and plant stress resistance, and various indicators of peony vegetative growth have been significantly reduced; The growth indicators were worse than those in group ③, which further indicated that the nutrients in mixture D were not as easily absorbed and utilized by peonies as in group ③; in group ⑤, spraying branches and leaves with mixture G could prevent the occurrence of fungal diseases of tree peony to a certain extent, but the nutrients were insufficient , reducing the stress resistance of the plant. Compared with ⑦ or ② and ⑧, the chlorophyll content is higher and the photosynthesis is enhanced, indicating that the plant is more robust and the disease resistance is enhanced. Based on the above analysis, the special bacterial fertilizer for peony can provide rich nutrients, promote photosynthesis and vegetative growth of peony, improve the stress resistance of plants, and effectively prevent the occurrence of fungal diseases of peony.

In summary, compared with the four groups ②, ③, ④ and ⑥, the chlorophyll content of group ② is the highest, the photosynthetic rate is the strongest, the average flowering period is the longest, and the number of susceptible plants is the least, which shows that the method provided by Example 1 The prepared bacterial fertilizer can promote the growth of peony roots, prolong the flowering period, and also have a certain control effect on peony fungal diseases; compared with the three groups ①, ② and ⑤, the peony in group ① has the best growth, the longest average flowering period, the longest new branches, and the corolla. It is huge, with a flowering rate of 100% (meaning that all new branches on the upper part of the plant can form flowers), and only one or two peonies are infected with the disease, indicating that the use of bacterial fertilizers combined with Bacillus subtilis HS5B5 bacterial agent has a better effect on preventing fungal diseases on peonies; ① and ⑦ Or ② compared with ⑧, it shows that the peony special bacterial fertilizer prepared in Example 1 is more targeted than common microbial bacterial fertilizer, and has good effect on the vegetative growth of peony and prolonging the flowering period.

Claims (10)

1. the preparation method of a Paeonia suffruticosa pouring root composite microbic bacterial fertilizer, it is characterised in that comprise the following steps:

Step one, take candida tropicalis and activate, cultivate to exponential phase, obtain the mixture containing candida tropicalis A；1:4 in mass ratio takes sweet potato waste and fruit garbage, is crushed to below 1.5cm, with sweet potato waste and fruit garbage gross mass On the basis of, add the carbamide of 1%, be uniformly mixing to obtain mixture B；Being inoculated in mixture B by mixture A, inoculum concentration is mixing The 1% of thing B mass, is stirred at room temperature fermentation 4-5 days, prepares fermented product I；

Step 2, in mass ratio 1:1:2-3:2-3 take chicken manure, cattle manure, biogas scum and are crushed to the wheat stalk of below 1cm, Mix homogeneously obtains mixture C, on the basis of the quality of mixture C, adds the quick lime of 5%, is exposed in air, naturally puts Put 3-5 days, prepare fermented product II；

Step 3, the fermented product II that step 2 prepares is joined in the fermented product I that step one prepares, wherein fermented product II with send out Ferment thing I mass ratio is 3:1, obtains mixture D after mixing；In mixture D, inoculation is in the geotrichum candidum of exponential phase, huge Bacillus cereus and bacillusmusilaginosiengineering, and the inoculum concentration of each bacterial strain is the 1% of mixture D quality, stirs under the conditions of 30 DEG C Mix fermentation 4-6 days, prepare fermented product III；

Step 4, with Herba Medicaginis as host plant, expanding propagation Glomus intraradices in expanding propagation substrate, inoculum concentration is 3%-5%, when ball in root When the mould spore of capsule reaches to contain more than 30 in every gram of soil, smash Radix Medicaginis and expanding propagation substrate, it is thus achieved that containing Glomus intraradices mycorhiza Mycelia and infect the mixture E 1 of root segment inside and outside fungal spore, root；With Herba Medicaginis as host plant, expanding propagation Moses in expanding propagation substrate Sacculus is mould, and inoculum concentration is 3%-5%, when during Glomus mosseae spore reaches every gram of soil containing more than 30, smashes Radix Medicaginis and expansion Numerous substrate, it is thus achieved that containing mycelia inside and outside Glomus mosseae mycorrhizal fungal spore, root and the mixture E 2 that infects root segment；By mixture E1 and mixture E 2 mix homogeneously of 1:1 in mass ratio obtain mixture E；

Step 5, in mass ratio 100:90:2:1 take fermented product III, mixture E, fish flour and zinc sulfate, by mixture E, fish flour and Zinc sulfate joins in fermented product III, prepares the mixture F of semi-solid after mixing；

Step 6, take bacillus subtilis HS5B5, activate in seed culture medium, proceed to after activation fermentation medium ferments, When bacillus subtilis HS5B5 grows to stable phase, obtain the mixture G containing bacillus subtilis HS5B5；

The mixture G 3-5:1 in mass ratio that step 7, mixture F step 5 prepared prepare with step 6 mixes, and stirring is all Even i.e. prepare Paeonia suffruticosa pouring root composite microbic bacterial fertilizer.

The preparation method of a kind of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer the most according to claim 1, it is characterised in that: step Fruit garbage described in rapid one be in Fructus Mali pumilae shedding, Fructus Mali pumilae corruption fruit, the fresh slag of Fructus Mali pumilae, pineapple bran any one or arbitrarily mix Thing.

The preparation method of a kind of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer the most according to claim 2, it is characterised in that: step Fruit garbage described in rapid one is the fresh slag of Fructus Mali pumilae and the mixture of Fructus Mali pumilae shedding, and the fresh slag of Fructus Mali pumilae and the mass ratio of Fructus Mali pumilae shedding For 1:3.

The preparation method of a kind of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer the most according to claim 1, it is characterised in that: step Sweet potato waste described in rapid one and fruit garbage are crushed to below 0.8cm.

The preparation method of a kind of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer the most according to claim 1, it is characterised in that: step Described in rapid two, the mass ratio of chicken manure, cattle manure, biogas scum and wheat stalk is 1:1:2:3.

The preparation method of a kind of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer the most according to claim 1, it is characterised in that: step Expanding propagation substrate used by rapid four expanding propagation Glomus intraradices and Glomus mosseae is gone out after being effluent sand and soil 1:1 in mass ratio mixing Bacterium prepares.

The preparation method of a kind of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer the most according to claim 1, it is characterised in that: step Seed culture medium described in rapid six is nutrient broth medium.

The preparation method of a kind of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer the most according to claim 7, it is characterised in that: step The activation temperature of the bacillus subtilis HS5B5 described in rapid six is 30 DEG C, during activation a length of 11 hours.

The preparation method of a kind of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer the most according to claim 1, it is characterised in that: step Fermentation medium described in rapid six is often to add 1000ml water as reference, and its component and addition thereof are respectively as follows: Semen Maydis powder 15.0g, glucose 3.0g, Semen sojae atricolor powder 15.0g, peptone 3.0g, MgSO₄·7H₂O 0.5g, KH₂PO₄·2H₂O 1.0g, water 1000ml, regulates pH to 7.0.

The preparation method of a kind of Paeonia suffruticosa pouring root composite microbic bacterial fertilizer the most according to claim 9, it is characterised in that: The condition that bacillus subtilis HS5B5 described in step 6 ferments in the fermentation medium is: 30 DEG C of aerobic fementations 25 hours.