CN104230563B - Microbial organic fertilizer containing bacillus cereus - Google Patents

Abstract

The invention particularly relates to a microbial organic fertilizer containing bacillus cereus, which comprises 1-5% of bacillus cereus liquid, 35-40% of tobacco residue, 35-45% of peat, 8-10% of oil cake and 9-12% of flower residue, wherein the raw materials are uniformly mixed, added with water accounting for 10% of the total volume, and subjected to stacking fermentation. Turning the piles once every 5 days, and fermenting for 40 days to obtain the biological organic fertilizer, wherein the effective active bacteria (bacillus cereus) of the bacterial fertilizer is more than or equal to 0.5 hundred million/g (cfu). The biological organic fertilizer is applied as a base fertilizer, and the bacillus cereus zymocyte liquid is diluted by 5000 times, and is irrigated to the root of a tobacco plant or sprayed to a page after being transplanted, so that the tobacco black shank can be effectively prevented and controlled, and the environmental pollution caused by the use of chemical agents is reduced.

Description

A kind of microbial organic fertilizer containing Bacillus cereus

technical field

The invention belongs to the technical field of biopesticides, and relates to a bio-organic fertilizer containing Bacillus cereus. Specifically, the microbial fertilizer produced by Bacillus cereus is combined with the fermentation liquid of the bacteria to jointly inhibit the occurrence of tobacco black shank.

Background technique

Tobacco black shank (Phytophthora parasitica var.nicotiane) is a devastating disease of tobacco, and it is also one of the most serious diseases in the world's tobacco production. pervasive and severe. In recent years, due to the expansion of continuous cropping tobacco fields and the increase of continuous cropping years, tobacco black shank disease has become more prevalent in many places. Over the years, in order to reduce the losses caused by the disease, people have tried various methods such as agricultural control, chemical control, and disease-resistant breeding to control tobacco black shank. The post-plant incubation period is short, the onset is fast, and multiple re-infections can occur in one growing season. At the same time, the disease sometimes occurs in combination with other diseases, resulting in the loss of resistance of some blackleg resistant varieties; moreover, the disease resistance Large-scale single planting of varieties in consecutive years, unreasonable layout, easy to lose the resistance of good varieties, shorten the service life of disease-resistant varieties, and once the disease is prevalent, it is difficult to control. At present, most of the chemical agents used to prevent and control tobacco black shank in production are phenylamides and fosfo-aluminum or several systemic fungicides with the same mechanism of action, which are prone to drug resistance and cause drug resistance. Efficacy decreased and dosage increased. Tobacco is an economic crop for leaves, which has special requirements on the appearance and internal quality of tobacco leaves. Pesticide residues, resistance of pathogenic bacteria and impact on cigarette hygiene greatly limit the large-scale use of pesticides. Under the trend of tobacco and tobacco products gradually developing towards pollution-free direction, biological control has been paid more and more attention as a safe and effective method.

The antagonistic bacteria of tobacco black shank bacteria include Pseudomonas spp. and Ba-cillus spp., which mainly dissolve the mycelia of tobacco black shank fungus by producing antibiotics; Bacteria found to significantly reduce the chances of tobacco black shank infection, thereby reducing the incidence of disease. The main method adopted at present is to make the antagonistic bacteria into concentrated liquid preparation, pure spore powder or composite solid powder, and then apply them before transplanting the tobacco seedlings. These methods are relatively simple, because the biocontrol strains are affected by complex soil ecological factors, the stability of the control effect of the biocontrol agent cannot be guaranteed, and the control effect is not obvious. What this patent adopts is the method of "combined drug use". First, the base fertilizer is applied, and then the biocontrol bacteria can be colonized in the soil through multiple sprays and finally form a dominant flora, thereby preventing tobacco black shank. to a better control effect.

Contents of the invention

The purpose of the invention is to solve the deficiency of the existing technology of using a single antagonistic agent (liquid) to prevent and treat tobacco black shank. Aiming at providing a bio-organic fertilizer containing Bacillus cereus, the new technology of combining the antibacterial fertilizer with antagonistic bacteria to prevent and control diseases can improve the colonization ability of antagonistic bacteria in soil and effectively improve its ability to Disease resistance. It is of great significance to improve the microenvironment of tobacco fields and promote the development of biological control technology.

The present invention is realized through the following technical solutions:

A bio-organic fertilizer containing Bacillus cereus, comprising 1%-5% of Bacillus cereus liquid, 35%-40% of soot residue, 35%-45% of peat, and 8% of oil bacillus by mass percentage ～10%, flower residue 9%～12%, preferably Bacillus cereus bacterial liquid 1%, smoke residue 40%, peat 40%, oil 10%, flower residue 9%.

The Bacillus cereus is fermented and cultivated in a liquid culture medium, the composition of which is: glucose 2.0g/L-2.8g/L, malt extract 2.0g/L-2.5g/L, peptone 1.0g/L ～1.2g/L, NaCl1.0mg/L～1.5mg/L, (NH4) ₂ SO ₄ 1.0mg/L～1.5mg/L, anhydrous CaCl ₂ 0.1mg/L～0.5mg/L, ZnSO ₄ · 7H ₂ O0.1mg/L～0.5mg/L, FeCl3 _{· 6H2O0.01mg} /L～0.05mg/L, pH6.5～7.2. Preferably, the medium components are: glucose 2.0g/L, malt extract 2.0g/L, peptone 1.0g/L, NaCl 1.2mg/L, (NH4) ₂ SO ₄ 1.5mg/L, anhydrous CaCl ₂ 0.2 mg/L, ZnSO ₄ ·7H ₂ O0.3mg/L, FeCl3 _{· 6H2O0.02mg} /L, pH6.8, _H2O1000m1 .

The liquid culture medium is prepared by the following method: Weigh the above-mentioned components and dissolve them in 1L of distilled water, adjust the pH to 6.8 with a pH meter after dissolving, put the above-mentioned solutions in a triangular flask and place them in a sterilizing pot at Sterilize at 121°C for 20 minutes. After the culture medium is cooled, dip 1ml of Bacillus cereus into the culture medium with an inoculation loop, and culture it on a shaker at 37°C at 100rpm for 48hrs.

The glucose can also be sucrose;

The peptone can also be replaced by corn steep liquor or tryptone.

The bacillus cereus colonies protrude on the LB solid medium, with smooth and fine edges, milky white, and a diameter of 1.1 mm to 1.8 mm.

The microbial organic fertilizer is prepared by the following method: uniformly mix the Bacillus cereus bacterial liquid with peat moss, oil dryness, flower residue and carbon ammonia water respectively, add 10% of the total volume of water, and carry out heaping, every 5 days The bio-organic fertilizer is obtained by turning the compost once and fermenting for 40 days.

The use of the microbial organic fertilizer for jointly inhibiting tobacco black shank is as follows: first, the biological organic fertilizer is applied as a base fertilizer, 80g-100g per pond, and 90kg-110kg per mu. At the same time, dilute the Bacillus cereus fermented bacterial liquid by 4000-5500 times, and water the roots of the tobacco plants after transplanting, with an amount of 9kg-12kg per mu. In the seedling stage of tobacco plant growth, watering is used when it is relatively sparse, and sprinkler irrigation is used when tobacco leaves are dense in the later stage of tobacco plant growth. Use a sprinkler system to spray on the leaf surface of the tobacco plant, and the leaves and roots must be soaked every time it is sprayed, once every 15 days, and twice a month.

Beneficial effects of the present invention: The microbial organic fertilizer of the present invention has positive significance for the growth-promoting effect of tobacco. At the same time, the joint application of the bacterial liquid of the biological bacteria can effectively prevent tobacco black shank, achieve no chemical pesticides and can effectively inhibit tobacco. The occurrence of black shank disease or the reduction of the degree of harm, this technology meets the requirements of the current transformation of agricultural production to green, environmental protection and sustainable development, and it is also important for improving the safety of tobacco and tobacco products and reducing the harm caused by smoking to health. is of great significance.

Detailed ways

The present invention will be further described below in conjunction with the embodiments. The following descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention to other forms. Changes or modifications are equivalent embodiments with equivalent changes. Any simple modifications, equivalent changes and modifications made to the above embodiments according to the technical essence of the present invention without departing from the solution content of the present invention all fall within the protection scope of the present invention.

Example 1

A bio-organic fertilizer containing Bacillus cereus, which consists of 1kg of Bacillus cereus bacterial liquid, 35kg of soot residue, 45kg of peat, 8kg of oil soot and 9kg of flower residue.

The bio-organic fertilizer is prepared by the following methods:

1) Weigh glucose 2.0g/L, malt extract 2.0g/L, peptone 1.2g/L, NaCl 1.0mg/L, (NH4) ₂ SO ₄ 1.5mg/L, anhydrous CaCl ₂ 0.3mg/L , ZnSO ₄ 7H ₂ O0.5mg/L, FeCl ₃ 6H ₂ O0.01mg/L, pH6.9;

2) Dissolve the above components in 1L of distilled water, and adjust the pH to 6.8 with a pH meter after dissolution;

3) Put the above solution in a triangular flask and sterilize it at 121°C for 20 minutes in a sterilizing pot. After the culture medium is cooled, dip 1ml of Bacillus cereus into the medium with an inoculation loop and transfer it to the culture medium at 37°C. The shaker was shaken at 100 rpm for 48 hours.

4) Uniformly mix the prepared Bacillus cereus bacterial liquid with peat carbon, oil dryness, flower residue and carbon ammonia water respectively, add 10% of the total volume of water, and carry out heap covering, turn over the heap every 5 days, and ferment for 40 Days later, the bio-organic fertilizer was prepared, and the effective active bacteria (Bacillus cereus) of the bacterial fertilizer was more than or equal to 0.5 billion/g (cfu).

Example 2

A bio-organic fertilizer containing Bacillus cereus, which consists of 4kg of Bacillus cereus bacterial liquid, 39kg of soot residue, 35kg of peat, 10kg of oil soot and 12kg of flower residue.

The bio-organic fertilizer is prepared by the following methods:

1) Weigh glucose 2.3g/L, malt extract 2.4g/L, peptone 1.0g/L, NaCl 1.3mg/L, (NH4) ₂ SO ₄ 1.5mg/L, anhydrous CaCl ₂ 0.2mg/L , ZnSO ₄ 7H ₂ O0.1mg/L, FeCl ₃ 6H ₂ O0.03mg/L, pH7.0, H ₂ O1000m1;

2) Dissolve the above components in 1L of distilled water, and adjust the pH to 6.8 with a pH meter after dissolution;

3) Put the above solution in a triangular flask and sterilize it at 121°C for 20 minutes in a sterilizing pot. After the culture medium is cooled, dip 1ml of Bacillus cereus into the medium with an inoculation loop and transfer it to the culture medium at 37°C. The shaker was shaken at 100 rpm for 48 hours.

4) Uniformly mix the prepared Bacillus cereus bacterial liquid with peat carbon, oil dryness, flower residue and carbon ammonia water respectively, add 10% of the total volume of water, and carry out heap covering, turn over the heap every 5 days, and ferment for 40 Days later, the bio-organic fertilizer was prepared, and the effective active bacteria (Bacillus cereus) of the bacterial fertilizer was more than or equal to 0.5 billion/g (cfu).

Example 3

A bio-organic fertilizer containing Bacillus cereus, which consists of 1kg of Bacillus cereus bacterial liquid, 40kg of soot residue, 40kg of peat, 10kg of oil soot and 9kg of flower residue.

The bio-organic fertilizer is prepared by the following methods:

1) Weigh glucose 2.0g/L, malt extract 2.0g/L, peptone 1.0g/L, NaCl 1.2mg/L, (NH4) ₂ SO ₄ 1.5mg/L, anhydrous CaCl ₂ 0.2mg/L , ZnSO ₄ 7H ₂ O0.3mg/L, FeCl ₃ 6H ₂ O0.02mg/L, pH6.8, H ₂ O1000m1;

2) Dissolve the above components in 1L of distilled water, and adjust the pH to 6.8 with a pH meter after dissolution;

3) Put the above solution in a triangular flask and sterilize it at 121°C for 20 minutes in a sterilizing pot. After the culture medium is cooled, dip 1ml of Bacillus cereus into the medium with an inoculation loop and transfer it to the culture medium at 37°C. The shaker was shaken at 100 rpm for 48 hours.

4) Uniformly mix the prepared Bacillus cereus bacterial liquid with peat carbon, oil dryness, flower residue and carbon ammonia water respectively, add 10% of the total volume of water, and carry out heap covering, turn over the heap every 5 days, and ferment for 40 Days later, the bio-organic fertilizer was prepared, and the effective active bacteria (Bacillus cereus) of the bacterial fertilizer was more than or equal to 0.5 billion/g (cfu).

Embodiment 4 microbial organic fertilizer detects and identifies

The biological fertilizer was prepared according to the formula in Example 3. Under the same conditions, the biological fertilizer containing Bacillus cereus was tested against the traditional organic fertilizer, and the content and activity of each component of the two fertilizers were compared.

Table 1 Detection data of biological fertilizer containing Bacillus cereus and traditional organic fertilizer

As can be known from Table 1, the nitrogen content of the Bacillus cereus-containing biological fertilizer of the present invention is higher than that of traditional organic fertilizers, and the total fertilizer is also not much different. Within the scope of plant absorption, other trace components also fully meet the requirements of fertilization. Requirements, but because biological fertilizers are more easily absorbed by the soil during implementation, their fertilizer efficiency is better than organic fertilizers.

Embodiment 5 containing bacillus cereus biological fertilizer is used for the effect of tobacco

This experiment was arranged in Yuxi City, Yunnan Province. The experimental biological fertilizer was developed and provided by the applicant. The control fertilizer was a conventional organic compound fertilizer on the market.

Fertilizer money is used to investigate the condition of soil black shank, select two test sites with similar conditions, use biological fertilizers and organic compound fertilizers according to a certain amount, investigate the disease index and calculate the control effect 30 days and 100 days after fertilization. The experimental results are as follows:

Table 2 Control effect of biological fertilizer containing Bacillus cereus on tobacco black shank

It can be seen from Table 2 that biological fertilizers can effectively prevent and control tobacco black shank, and the highest control effect can reach 98.46%, which has great advantages compared with organic compound fertilizers. Secondly, biological fertilizers can improve the yield and product quality It has a great promotion effect, greatly promotes the income of farmers, and has market economic benefits.

Claims (3)

1. A microbial organic fertilizer containing Bacillus cereus, characterized in that: in terms of mass percentage, it comprises 1% to 5% of Bacillus cereus bacterial liquid, 35% to 40% of soot residue, and 35% to 45% of peat. %, 8%~10% of oily dryness, 9%~12% of flower dregs; The Bacillus cereus is fermented and cultured in a liquid medium, and the components of the medium are as follows: glucose 2.0g/L, malt extract 2.0g/L, peptone 1.0g/L, NaCl 1.2mg/L, (NH ₄ ) ₂ SO ₄ 1.5mg/L, anhydrous CaCl ₂ 0.2mg/L, ZnSO ₄ 7H ₂ O 0.3mg/L, FeCl ₃ 6H ₂ O 0.02mg/L, H ₂ O 1000mL; The microbial organic fertilizer is prepared by the following method: uniformly mix the Bacillus cereus bacterial liquid with peat, oil dryness, flower residue and carbon ammonia water respectively, add 10% of the total volume of water, and carry out heaping, once every 5 days After turning over and fermenting for 40 days, the bio-organic fertilizer is obtained. 2. a kind of microbial organic fertilizer containing Bacillus cereus according to claim 1 is characterized in that: by mass percent, comprising Bacillus cereus bacterium liquid 1%, soot residue 40%, peat 40%, Oil dry 10%, flower dregs 9%. 3. A microbial organic fertilizer containing Bacillus cereus according to claim 1 or 2, characterized in that: said microbial organic fertilizer is used for joint suppression of tobacco black shank.