CN110791459B

CN110791459B - Bacillus subtilis for preventing and controlling continuous cropping lily soil-borne blight and application thereof

Info

Publication number: CN110791459B
Application number: CN201911227973.8A
Authority: CN
Inventors: 黄启为; 沈其荣; 任胜林; 梅慧玲; 李静
Original assignee: Nanjing Agricultural University
Current assignee: GENLIDUO BIO-TECH Corp.,Ltd.
Priority date: 2019-12-04
Filing date: 2019-12-04
Publication date: 2020-09-08
Anticipated expiration: 2039-12-04
Also published as: CN110791459A

Abstract

The invention discloses a bacillus subtilis for preventing and controlling continuous cropping lily soil-borne wilt and application thereof, wherein the antagonistic bacterium HQWBH1 is obtained by separation, and is inoculated into a mixture of an organic fertilizer and acid hydrolyzed amino acid for solid fermentation to prepare a microbial fertilizer product, and the product contains 1 × 10 percent⁸The total nitrogen content of the antagonistic bacteria of more than cfu/g is 4-5 percent (more than 90 percent is organic nitrogen), the total nitrogen phosphorus potassium nutrient is 6-10 percent, and the organic matter content is more than or equal to 45 percent. The pot experiment result shows that the prevention and control rate of the lily soil-borne blight reaches over 75 percent, and if the fertilizer product is applied to soil where lily is planted in successive years, the lily soil-borne blight can be effectively controlled.

Description

Bacillus subtilis for preventing and controlling continuous cropping lily soil-borne blight and application thereof

Technical Field

The invention belongs to the field of agricultural microorganisms, and relates to bacillus subtilis for preventing and controlling continuous cropping lily soil-borne blight and application thereof.

Background

Crop blight is a typical soil-borne disease, which is extremely harmful and can cause crop failure in severe cases. Although crop blight is caused by infection of a pathogenic bacterium, namely fusarium oxysporum, fusarium oxysporum is caused by different specialization types of infection of the fusarium oxysporum. If watermelon fusarium wilt is caused by fusarium oxysporum watermelon specialization infection, cucumber fusarium wilt is caused by fusarium oxysporum cucumber specialization infection, sweet melon fusarium wilt is caused by fusarium oxysporum melon specialization infection, bitter melon fusarium wilt is caused by fusarium oxysporum bitter melon specialization infection, banana fusarium wilt is caused by fusarium oxysporum banana specialization middle-4 physiological race infection, lily fusarium wilt is caused by fusarium oxysporum lily specialized infection, and the like.

The lily wilt disease is also called stem rot and root rot, is the most common and serious disease in more than 20 disease types of lily, and is caused by dip-dyeing of soil-borne fungal pathogenic bacteria (fusarium oxysporum, fusarium moniliforme and fusarium solani). Among them, fusarium oxysporum lily specialization type is a main pathogenic bacterium, which seriously affects the yield and quality of lily. The lily wilt mainly damages underground tubers, the disease can be developed after 2-3 continuous cropping, and the more continuous cropping, the more serious the disease. The key problem of lily wilt is not nutrient deficiency or unbalance, nor secondary salinization of soil, but rather toxicity of microflora and secondary substances thereof in soil.

201210119093.0 discloses a bacillus subtilis and its application in preventing and treating cucumber fusarium wilt, the bacillus subtilis can inhibit cucumber specialization of cucumber fusarium oxysporum which is a pathogen of cucumber fusarium wilt, and the prevention effect on cucumber fusarium wilt in seedling stage can reach 58.66%. 201810488484.7 provides a new strain Bacillus subtilis hfkc07, which can be applied in preventing and treating watermelon fusarium wilt and banana fusarium wilt. 201410440682.8 discloses a strain for antagonizing cotinus coggygria fusarium oxysporum, which has obvious plate inhibition effect on the cotinus coggygria fusarium oxysporum. However, the pathogenic bacteria of the wilt disease are species-specific for infection of the host. Although the pathogenic bacteria are fusarium oxysporum, the specialization types of the pathogenic bacteria are different, and the types of crops causing the blight are different. For example, the pathogenic bacteria causing cucumber fusarium wilt are fusarium oxysporum cucumber specialization types, which cannot cause banana fusarium wilt. The pathogenic bacteria causing banana wilt are No. 4 physiological race in fusarium oxysporum banana specialization type, which cannot cause watermelon wilt or lily wilt. Because the antagonistic strains (i.e. antagonistic strains) of pathogenic bacteria of cucumber fusarium wilt have no inhibiting effect on pathogenic bacteria of fusarium wilt in other species, different antagonistic bacteria need to be screened for specialized pathogenic bacteria of fusarium wilt of different crops to realize effective prevention and control. On the other hand, the biocontrol strain is required to be capable of being practically applied to the preparation of biopesticides or biofertilizers and to be capable of being colonized at the rhizosphere of crops. The soil for continuous cropping of a certain crop usually contains the specific secretion of the crop, and a specific microbial environment is formed in the soil, so that foreign microorganisms are difficult to effectively colonize.

The biological organic fertilizer is a fertilizer which is compounded by microorganisms with specific functions and organic materials mainly prepared from animal and plant residues (such as crop straws, livestock and poultry manure and the like) through harmless treatment and decomposition. The biological organic fertilizer is essentially characterized by containing beneficial microorganisms with specific functions and showing certain fertilizer effects, and the vital activities of the specific beneficial microorganisms are key factors of the biological organic fertilizer being superior to other fertilizers. Numerous studies have shown that: the biological organic fertilizer plays a positive role in enhancing crop stress resistance, providing crop nutrients, increasing fertilizer efficiency, improving nutrient absorption of crops, regulating soil pH value and the like in agricultural production.

The Chinese medicine dregs contain rich organic and inorganic matters. Chinese medicine resources are rich, and medicine dregs generated after the effective components are extracted in a large scale every year are discarded everywhere, so that the environment is seriously polluted, and the raw materials which can be used as organic fertilizer products are greatly wasted. If the solid organic wastes are fermented at high temperature to prepare decomposed compost and then the decomposed compost is used as a carrier for preparing biological (containing specific beneficial microorganisms) inoculants, the application prospect is good.

Disclosure of Invention

The invention aims to provide a bacillus subtilis for preventing and controlling continuous cropping lily soil-borne blight aiming at the defects in the prior art.

Another purpose of the invention is to provide application of the bacillus subtilis.

The invention also aims to provide a special microbial organic fertilizer for preventing and controlling continuous cropping soil-borne blight and a preparation method thereof.

A Bacillus subtilis HQWBH1 for preventing and controlling continuous cropping lily soil-borne wilt is preserved in China general microbiological culture Collection center in 11 months and 01 days in 2019, and the strain preservation number is as follows: CGMCC NO. 18779.

The invention discloses application of bacillus subtilis HQWBH1 in preparation of a microbial organic fertilizer for preventing and controlling continuous cropping soil-borne wilt.

A preparation method of a microbial organic fertilizer special for preventing and controlling continuous cropping soil-borne blight is characterized in that fermentation liquor of bacillus subtilis HQWBH1 is inoculated into a mixed fermentation product of an organic fertilizer and full-value amino acid for solid secondary fermentation to obtain the microbial organic fertilizer.

Preferably, the preparation method of the bacillus subtilis HQWBH1 fermentation liquor comprises the following steps of inoculating antagonistic strain HQWBH1 into LB culture solution for liquid fermentation production, wherein the conditions of the fermentation production are that the pH is 7.0, the fermentation temperature range is 30-35 ℃, the stirring speed is 180-300 r/min, the fermentation time is 48hr, and the bacteria or spore content of the fermentation liquor is more than or equal to 1 × 10¹⁰One per ml.

Preferably, the solid secondary fermentation process comprises turning over the pile for 1 time per day at a temperature not higher than 50 deg.C for 5 days, and the content of HQWBH1 reaches 1 × 10⁸The water content is controlled to be below 30 ℃ per gram, and the microbial organic fertilizer for preventing and controlling continuous cropping soil-borne blight is obtained.

Preferably, the preparation method comprises the following steps: (1) automatically crushing livestock and poultry dead of diseases in a closed container, automatically transferring all solid matters and liquid into a closed hydrolysis tank, and then performing hydrolysis at an initial acid concentration c (1/2H)₂SO₄) Is 2.5 to 5mol L^-1Hydrolyzing at 80-100 deg.C under 1-2 atmospheric pressure for 2-5 hr; (2) standing for layering after the solution in the hydrolysis tank is cooled to below 80 deg.C, and collecting the amino acid solution in the middle layer; or the complete amino acid is an animal protein amino acid water-soluble fertilizer produced by Union biotechnology limited of Jiangyun city of Jiangsu province.

Preferably, the organic fertilizer is decomposed compost or organic fertilizer sold in the market; the decomposed compost is preferably decomposed straw compost and decomposed traditional Chinese medicine residue compost. The decomposed traditional Chinese medicine residue compost can be automatically decomposed by extracting waste materials from purchased medicine enterprises, and the types of traditional Chinese medicines in the traditional Chinese medicine residues are not limited.

The germination index of the mixed fermentation product of the organic fertilizer and the full-value amino acid is more than 98 percent, the content of organic matters is more than or equal to 50 percent, the content of free amino acid is more than or equal to 0.50 percent, the content of organic nitrogen is 1.2 to 2.0 percent, and the water content is 25 to 30 percent.

Preferably, the mixed fermentation product of the organic fertilizer and the full-value amino acid is prepared by the following method: (1) adding full-valence amino acid into the organic fertilizer according to 3-8% of the weight of the organic fertilizer, and fully and uniformly mixing; (2) turning for 1 time every 2 days, and continuously turning for 3-5 times to obtain the final product.

The microbial organic fertilizer for preventing and controlling continuous cropping soil-borne blight is prepared according to the preparation method.

The microbial organic fertilizer product preferably contains 1 × 10⁸The antagonistic bacteria HQWBH1 with the total nitrogen content of more than cfu/g is 4-5%, the total nitrogen phosphorus potassium nutrient content is 6-10%, and the organic matter content is more than or equal to 45%.

The microbial organic fertilizer disclosed by the invention is applied to the prevention and treatment of continuous cropping lily soil-borne blight and the improvement of lily yield.

Has the advantages that:

compared with the organic fertilizer products on the current market, the invention has the following advantages:

1) the invention screens a specific high-efficiency strain HQWBH1 for inhibiting the growth of the lily soil-borne wilt pathogenic fungi, and the inhibition effect is very obvious. The pot experiment result shows that the prevention and control rate of the lily soil-borne blight reaches over 78 percent, and if the fertilizer product is applied to soil where lily is planted in successive years, the lily soil-borne blight can be effectively controlled. The strain is derived from rhizosphere soil of healthy lily plants in lily continuous cropping fields, and after the microbial fertilizer prepared is applied to the lily continuous cropping soil, the microbial fertilizer can be well adapted to soil microbial environment and can be colonized in a large amount at the lily rhizosphere.

2) The microbial product contains rich organic matters (not less than 40%), the total nitrogen content is 4-5% (more than 90% of total nitrogen by mass is organic nitrogen), and most of the total nitrogen is amino acid and micromolecular polypeptide. The product has the total nitrogen, phosphorus and potassium nutrient mass ratio of 8-10%, the content of free amino acid not less than 0.3%, and particularly high biological effectiveness.

3) The application of the microbial product has no problems caused by the use of chemical pesticides, can avoid or reduce the use amount of other chemical pesticides for preventing and controlling lily wilt, and is beneficial to the pollution-free production of lily.

4) Because the biological strain preparation is a biological strain preparation, a series of problems caused by the use of chemical pesticides are completely avoided, so that the pollution-free production of the lily is facilitated, farmers can not use or reduce the use amount of other chemical pesticides, the expenditure of the farmers can be saved, and the export of agricultural products is facilitated.

5) The microbial product also has the function of increasing yield, thereby not only saving the expenditure, but also increasing the income of farmers.

6) The microbial product not only has the function of an organic fertilizer, but also has the function of antagonistic bacteria contained in the organic fertilizer through various biocontrol mechanisms, thereby achieving the effect of preventing and controlling the blight transmitted by the soil.

Drawings

FIG. 1 antagonistic Effect of HQWBH1 Strain

A: growth of a plate with a special type of Fusarium oxysporum and Bulbus Lilii

B: the confronting effect of HQWBH1 strain and fusarium oxysporum lily specialized flat plate

Biological material preservation information

HQWBH1, classified and named as Bacillus subtilis, and deposited in China general microbiological culture Collection center, with the deposit address of the institute of microbiology, China academy of sciences, No. 3, West Lu No.1 institute of North Chen, west way, the south facing Yang, Beijing, the date of deposit is 2019, 11 months and 01 days, and the strain deposit number is: CGMCC NO. 18779.

Detailed Description

Example 1

(I) isolation of antagonistic strains

Primary screening of antagonistic strains:

collecting rhizosphere soil of healthy lily plants in lily continuous cropping fields, adding Fusarium oxysporum lily specialized (Fusarium oxysporum sp.f.liii) which is a pathogen of blight and is capable of transmitting blight to lily soil by adopting PDA + NA, separating and purifying the diseased plants collected from the fields with serious blight and blight diseases by Jiangsu Yixing shu lily soil, primarily screening antagonistic strains by using a double-layer flat plate of spore suspension, primarily screening 30 antagonistic strains, freeze-drying the strains at-20 ℃, and storing for further screening.

Re-screening antagonistic strains:

taking the preliminarily screened antagonistic strains and pathogenic bacteria, carrying out a confrontation test on a PDA (personal digital assistant) plate, and carrying out rescreening. A piece of pathogenic bacteria block is dug from the flat plate full of pathogenic bacteria to the center of the PDA flat plate by a puncher (5mm), and the PDA flat plate is placed in an incubator at 28 ℃ for culture. Inoculating the antagonistic bacteria to be tested in the center of the PDA plate after 1d (24h) and 2cm away from the periphery of the pathogenic bacteria block with a sterilized toothpick, culturing at 28 ℃ for 3-5 d, and observing the inhibition condition of the bacterial colony of the antagonistic bacteria to the pathogenic bacteria. The final screening yielded the strain HQWBH1, the antagonistic effect of which is shown in fig. 1.

Identification of antagonistic strains

The HQWBH1 is characterized by gram-positive thallus, sprout-producing arms, elliptical shape of the sprout arms, expanded bag, aerobic growth, milk peptonization, hydrolysis of starch and casein, reduction of nitrate, no hydrolysis of tyrosine, and positive in catalase test, dihydroxyacetone from glycerol test and VP test; MR is negative and can utilize fructose, cellobiose, mannose, glucose, sucrose and citrate. The antagonistic strain HQWBH1 was identified as Bacillus subtilis based on 16S rRNA, with Genbank number MN611445 for the 16S rRNA gene sequence.

EXAMPLE 2 microbial Fertilizer production

(1) Inoculating an antagonistic strain HQWBH1 into LB culture solution for liquid fermentation production, wherein the conditions of the fermentation production are that the pH is 7.0, the fermentation temperature range is 30-35 ℃, the stirring speed is 180-300 r/min, the fermentation time is 48hr, and the bacteria or spore content of the fermentation liquor is more than or equal to 1 × 10¹⁰One per ml.

(2) Inoculating 50L/ton fermentation liquid of HQWBH1 into decomposed amino acid Chinese medicinal residue compost (mixed fermentation product of Chinese medicinal residue compost and full-value amino acid) for solid secondary fermentationTurning over and piling for 1 time every day during fermentation at a temperature not higher than 50 deg.C for 5 days, and making the HQWBH1 content reach 1 × 10⁸The microbial fertilizer 1 containing the antagonistic strain HQWBH1 is prepared by controlling the water content to be below 30 ℃ per gram. The germination index of the decomposed traditional Chinese medicine residue compost is more than 98%, the content of organic matters is more than or equal to 50%, the content of free amino acids is more than or equal to 0.50%, the content of organic nitrogen is 1.2-2.0%, and the water content is 25-30%.

The full-value amino acid is directly purchased from the animal protein amino acid water-soluble fertilizer produced by the Peng ray river cloudy city biotechnology limited company.

The amino acid traditional Chinese medicine residue compost is prepared by the following method: (1) adding full-value amino acid into the thoroughly decomposed traditional Chinese medicine residue compost according to 3-8% of the weight of the thoroughly decomposed traditional Chinese medicine residue compost, and fully and uniformly mixing; (2) turning the pile for 1 time every 2 days, and continuously turning the pile for 3-5 times.

The special compound microbial fertilizer for preventing and controlling the blight transmitted by the soil contains 1 × 10⁸The mass ratio of the total nitrogen content of HQWBH1 of more than one/g is 4-5%, the mass ratio of more than 90% of the total nitrogen is organic nitrogen, the mass ratio of total nitrogen, phosphorus and potassium nutrients is 8-10%, the content of free amino acid is not less than 0.3%, and the mass ratio of the organic matter content is not less than 40%.

EXAMPLE 3 microbial Fertilizer production

(2) Inoculating 50L/ton fermentation liquid of strain HQWBH1 into decomposed amino acid corn straw compost (mixed fermentation product of straw compost and full-value amino acid) for solid secondary fermentation, turning over for 1 time every day at a temperature not higher than 50 deg.C for 5 days, and allowing the content of strain HQWBH1 to reach 1 × 10⁸The water content is controlled to be below 30 ℃ per gram, and the compound microbial fertilizer 2 containing the antagonistic strain HQWBH1 is obtained. The germination index of the decomposed straw compost is more than 98 percent, and the content of organic matters is more than or equal to50 percent, more than or equal to 0.50 percent of free amino acid, 1.2 to 2.0 percent of organic nitrogen and 25 to 30 percent of water content.

The full-valence amino acid is prepared by the following method: (1) automatically crushing livestock and poultry dead of diseases in a closed container, automatically transferring all solid matters and liquid into a closed hydrolysis tank, and then performing hydrolysis at an initial acid concentration c (1/2H)₂SO₄) Is 2.5 to 5mol L^-1Hydrolyzing at 80-100 deg.C under 1-2 atmospheric pressure for 2-5 hr; (2) and (3) standing for layering after the hydrolysis is finished and the solution in the hydrolysis tank is cooled to below 80 ℃, and collecting the amino acid solution in the middle layer.

The amino acid corn straw compost is prepared by the following method: (1) adding full-value amino acid into the decomposed corn straw compost according to 5% of the weight of the decomposed corn straw compost, and fully and uniformly mixing; (2) turning the pile for 1 time every 2 days, and continuously turning the pile for 3-5 times.

EXAMPLE 4 microbial Fertilizer production

(2) Inoculating 50L/ton fermentation liquid of strain HQWBH1 into decomposed amino acid pig manure compost (mixed fermentation product of pig manure compost and full-value amino acid) for solid secondary fermentation, turning over for 1 time each day during fermentation at a temperature not higher than 50 deg.C for 5 days, and allowing the content of strain HQWBH1 to reach 1 × 10⁸The water content is controlled to be below 30 ℃ per gram, and the compound microbial fertilizer 3 containing the antagonistic strain HQWBH1 is obtained. The germination index of the decomposed pig manure compost is more than 98 percent, the organic matter content is more than or equal to 50 percent, and the decomposed pig manure compost is freeAmino acid not less than 0.50%, organic nitrogen content of 1.2-2.0%, and water content of 25-30%.

The amino acid pig manure compost is prepared by the following method: (1) adding complete amino acid into the decomposed pig manure compost according to 8% of the weight of the decomposed pig manure compost, and fully and uniformly mixing; (2) turning the pile for 1 time every 2 days, and continuously turning the pile for 3-5 times.

Example 5 potted plant control efficacy test

The experiment was set up with 4 treatments: treatment 1 (CF): applying a fertilizer without applying a compound microbial fertilizer; and (3) treatment 2: application of composite microbial fertilizer 1 containing HQWBH1 strain (prepared in example 2); and (3) treatment: application of composite microbial fertilizer 2 containing HQWBH1 strain (prepared in example 3); and (4) treatment: composite microbial fertilizer 3 (prepared in example 4) containing HQWBH1 strain was applied. Three replicates were used.

Transplanting the grown lily into lily soil-borne blight continuous cropping soil (pathogen spore of lily soil-borne blight reaches 1.7 × 10⁴cfu/g soil), and the compound microbial fertilizer is applied in the treatment 2 according to the dosage of 1 percent of the weight of the soil. The dosage of all the treated nitrogen, phosphorus and potassium is consistent, and the conventional management is carried out. Then, the disease incidence is observed, the disease incidence is recorded, and whether the antagonistic strain has the effect of inhibiting lily wilt is determined.

After the lily seedlings are transplanted for 38 days, plants are found to be attacked in the control (CF, the compound microbial fertilizer is not applied), and the lily seedlings are allowed to grow continuously, namely 85% of the plants are attacked in the control 60 days after the lily seedlings are transplanted. The incidence and control effect results of the blight of the plants 60 days after the lily seedlings are transplanted are shown in table 1. The pot experiment result shows that: by applying the compound microbial fertilizer containing HQWBH1, the number of bacteria and fungi for preventing the linear diseases in the soil for continuous cropping of the lily can be obviously increased, the number of pathogenic fungi is obviously reduced (Table 1), the occurrence of the soil-borne blight of the lily can be effectively prevented and controlled, and the prevention effect is more than 77% (Table 2).

TABLE 1 influence of the specific compound microbial fertilizer on the microbial system of lily continuous cropping blight soil

Note: 1: comparison: chemical fertilizer; 2: a composite microbial fertilizer 1 (prepared in example 2) containing an antagonist against soil-borne blight of lily (HQWBH 1); 3: a composite microbial fertilizer 2 (prepared in example 3) containing an antagonist against soil-borne blight of lily (HQWBH 1); 4: a composite microbial fertilizer 3 (prepared in example 4) containing an antagonist against soil-borne blight of lily (HQWBH 1). All the soil to be tested was the lily soil-borne blight continuous cropping soil (the same below).

TABLE 2 Effect of the compound microbial fertilizer on prevention and control of lily wilt

Example 6 field biological assay

In 2018, a lily biological effect test is arranged in Yixing city of Jiangsu province in 9 months. Set 4 treatments: treatment 1 (CF): applying a fertilizer without applying a compound microbial fertilizer; and (3) treatment 2: application of composite microbial fertilizer 1 containing strain HQWBH1 (prepared in example 2); and (3) treatment: application of composite microbial fertilizer 2 containing HQWBH1 strain (prepared in example 3); and (4) treatment: composite microbial fertilizer 3 (prepared in example 4) containing HQWBH1 strain was applied. The three times of the treatment are repeated, and the total amount of nitrogen, phosphorus and potassium of the four treatments is consistent.

Before planting lily, spreading special compound microbial fertilizer for preventing and controlling blight transmitted by lily soil according to the amount of 400 kg/mu, carrying out rotary tillage by using a rotary cultivator for 30 centimeters of soil, fully and uniformly mixing, ridging (the width is 1.8 meters), and ditching (the width is 60 centimeters and the depth is 50 centimeters). Planting lily seeds on the ridges according to the row spacing of 30 cm, the plant spacing of 15-18 cm and the depth of 15 cm, covering soil, and performing conventional water and fertilizer management.

The total application amount of the compound fertilizer is 50-100 kg/mu: spreading 15-30 kg/mu of compound fertilizer (16-16-16) before overwintering (11 months bottom), and covering soil (1-2 cm); before sprouting (2-3 months in the next year), 35-70 kg/mu of compound fertilizer (16-16-16) is broadcast and covered with soil (1-2 cm).

Yield in 2019, 8 and 24 months, and the results (shown in Table 3) show that: the application of the compound microbial fertilizer can obviously increase the yield of the field lily, and the yield is increased by 13.9% compared with a control. 1000 lily bulbs were randomly picked and weighed one by one (table 4) and found: after the fertilizer is applied, the bulbs of the lily are small, and statistics shows that the number of the bulbs not exceeding 125 g accounts for 55%, and the number of the bulbs exceeding 125 g accounts for 45% (wherein no bulbs exceeding 150 g); treatment 2 (application of compound microbial fertilizer 1, prepared in example 2) had a bulb count ratio of 15% without more than 125 g and a bulb count ratio of 85% with more than 125 g (of which the bulb count of more than 150 g was 25%); treatment 3 (application of compound microbial fertilizer 2, prepared in example 3) had a bulb count ratio of 14.9% without exceeding 125 g and a bulb count ratio of 85.1% with exceeding 125 g (of which the bulb count of exceeding 150 g accounted for 24.6%); the treatment 4 (application of the compound microbial fertilizer 3, prepared in example 4) had a bulb count ratio of 14.2% without exceeding 125 g and a bulb count ratio of 85.8% with exceeding 125 g (wherein the bulb count of exceeding 150 g was 24.9%).

Table 3 influence of application of compound microbial fertilizer on yield of field lily (tea town in lixing city, Jiangsu province, 8 months and 24 days in 2019)

Table 4 influence of the application of the compound microbial fertilizer on the bulb weight of lily in the field (tea town in lixing city, Jiangsu province, 8 months in 2019 and 24 days in 2019)

According to the purchase price of farmers: when the weight of lily bulbs of less than 125 grams is 10 yuan per kilogram, the weight of lily bulbs of more than 125 grams is 15 yuan per kilogram, and the average yield per mu is 8000 yuan, the yield value of the treatment 2 (applying the compound microbial fertilizer 1, prepared in example 2) is 8045.5 yuan per mu, the yield value of the treatment 2 is 11545.5 yuan, the income per mu is increased by 3500 yuan, the cost of the compound microbial fertilizer is deducted to be 800 yuan per mu, and farmers can obtain 2700 yuan per mu (the net income of the treatment 3 and the treatment 4 is more than 2700 yuan per mu, because the proportions of the lily bulbs of the two treatments of more than 125 grams are higher than that of the treatment 2), so that the large-area application of the special compound microbial fertilizer for lily becomes possible.

The invention starts from the key problem of the soil microbial community (Table 1), develops a special compound microbial fertilizer product capable of remarkably preventing and controlling the occurrence of lily continuous cropping blight, and the biological prevention and control rate is more than 78%. The action mechanism of the product lies in that the effective carbon and the effective nitrogen source in the special product provide a good material basis for the cultivation of functional bacteria and soil microflora in the product, so that the ecology and the food chain of exogenous beneficial functional bacteria microorganisms are quickly established; on the other hand, the product contains higher content of organic nitrogen and organic phosphorus, and the nutrient substances are very beneficial to the growth of lily plants in the continuous cropping obstacle soil, thereby greatly improving the seedling establishment rate and the stress resistance of the lily plants.

Claims

1. A Bacillus subtilis HQWBH1 for preventing and controlling continuous cropping lily soil-borne wilt is preserved in China general microbiological culture Collection center in 11 months and 01 days in 2019, and the strain preservation number is as follows: CGMCC number 18779.

2. The application of the bacillus subtilis HQWBH1 in preparing the microbial organic fertilizer for preventing and controlling continuous cropping soil-borne blight.

3. A preparation method of a microbial organic fertilizer special for preventing and controlling continuous cropping soil-borne blight is characterized in that fermentation liquor of bacillus subtilis HQWBH1 of claim 1 is inoculated into a mixed fermentation product of an organic fertilizer and full-value amino acid for solid secondary fermentation to obtain the microbial organic fertilizer; the full-valence amino acid is prepared by the following method: (1) automatically crushing livestock and poultry dead of diseases in a closed container, automatically transferring all solid matters and liquid into a closed hydrolysis tank, and then performing hydrolysis at an initial acid concentration c (1/2H)₂SO₄) Is 2.5 to 5 mol.L^-1Hydrolyzing at 80-100 deg.C under 1-2 atmospheric pressure for 2-5 hr; (2) and (3) standing for layering after the hydrolysis is finished and the solution in the hydrolysis tank is cooled to below 80 ℃, and collecting the amino acid solution in the middle layer.

4. The preparation method of claim 3, wherein the fermentation broth of Bacillus subtilis HQWBH1 is prepared by inoculating antagonistic strain HQWBH1 into LB culture solution for liquid fermentation at pH7.0, fermentation temperature of 30-35 deg.C, stirring speed of 180-300 r/min, fermentation time of 48hr, and bacteria or spore content of the fermentation broth of not less than 1 × 10¹⁰One per ml.

5. The method of claim 3, wherein the solid secondary fermentation process comprises turning over the pile 1 time per day at a temperature not higher than 50 deg.C for 5 days, and the content of HQWBH1 reaches 1 × 10⁸The water content is controlled to be below 30 ℃ per gram, and the microbial organic fertilizer for preventing and controlling continuous cropping soil-borne blight is obtained.

6. The method according to claim 3, wherein the organic fertilizer is decomposed compost or a commercially available organic fertilizer.

7. The method according to claim 6, wherein the decomposed compost is decomposed straw compost, decomposed herb residue compost, and decomposed pig manure compost.

8. The method according to claim 3, wherein the mixed fermentation product of organic fertilizer and full-value amino acid is prepared by the following steps: (1) adding full-value amino acid into the organic fertilizer according to the weight of 3-8% of the organic fertilizer, and fully and uniformly mixing; (2) turning for 1 time every 2 days, and continuously turning for 3-5 times to obtain the final product.

9. The microbial organic fertilizer for preventing and controlling continuous cropping soil-borne blight, which is prepared by the preparation method of any one of claims 3-8.

10. The microbial organic fertilizer of claim 9, characterized in that said microbial organic fertilizer product comprises 1 × 10⁸The antagonistic bacteria HQWBH1 with the total nitrogen content of more than cfu/g is 4-5%, the total nitrogen phosphorus potassium nutrient content is 6-10%, and the organic matter content is more than or equal to 45%.

11. The use of the microbial organic fertilizer of claim 9 or 10 in the control of continuous cropping lily soil-borne wilt and in increasing lily yield.