Compound bacterial fertilizer for reducing soil obstacles as well as preparation method and application of compound bacterial fertilizer

Abstract

The invention discloses a compound bacterial fertilizer for reducing soil obstacles as well as a preparation method and application thereof, and relates to the technical field of biology. The compound bacterial fertilizer is prepared from a mortierella indurata bacterial agent, a trichoderma hedgehog bacterial agent, an osmanthus fragrans geotrichum bacterial agent and a bacillus cereus bacterial agent, the mortierella cinerea preparation is obtained by uniformly mixing a mortierella cinerea spore suspension and corn straw powder and then drying; the trichoderma hedgehog fungicide is obtained by uniformly mixing a trichoderma hedgehog spore suspension and corn straw powder and then drying; the osmanthus fragrans geotrichum fungicide is obtained by uniformly mixing osmanthus fragrans geotrichum spore suspension and corn straw powder and then drying, the bacillus cereus microbial inoculum is obtained by freeze-drying a bacillus cereus fermentation thallus. By applying the compound bacterial fertilizer to the surface layer of ginseng continuous cropping soil, the soil condition of the ginseng continuous cropping soil can be improved, ginseng growth is promoted, ginseng root rot is reduced, and the survival rate of ginseng plants is increased.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a composite bacterial fertilizer for reducing soil barriers, a preparation method and an application thereof. Background Art

[0002] Soil continuous cropping disorder, also known as replanting disease, is widely present in the cultivation of vegetables, Chinese herbal medicines and fruit trees. It refers to abnormal crop growth and development caused by continuous cultivation of the same crops or closely related crops on the same soil. Symptoms are generally poor growth and development, decreased yield and quality. In extreme cases, local seedling death, no seedlings or weak seedlings; the roots of most affected plants turn brown, branches decrease, vitality decreases, and the distribution range is narrow, resulting in a decreased ability to absorb water and nutrients. There are many reasons for the occurrence of continuous cropping disorder, including excessive nutrient consumption, deterioration of soil physical and chemical properties, increase in pests and diseases, and accumulation of toxic substances (including allelopathic substances). Its occurrence is affected by various environmental conditions, and the number of continuous cropping, soil properties and improper management of subsequent water and fertilizer will aggravate the disorder.

[0003] Ginseng (Panax ginseng CAmey.) is a perennial herbaceous plant of the Araliaceae family and the genus Panax. Planting ginseng is extremely sensitive to soil conditions and is prone to continuous cropping problems. The occurrence of continuous cropping problems for ginseng may lead to reduced production and income at the least, or even total crop failure at the worst, which seriously restricts the sustainable development of the ginseng industry. The present invention intends to develop a composite bacterial fertilizer to reduce soil continuous cropping problems. Summary of the invention

[0004] The purpose of the present invention is to provide a composite bacterial fertilizer for reducing soil barriers, a preparation method and an application thereof, so as to solve the problems existing in the above-mentioned prior art. By applying the composite bacterial fertilizer provided by the present invention on the surface of the soil of ginseng continuous cropping, the soil conditions of the ginseng continuous cropping land can be improved, the growth of ginseng can be promoted, the occurrence of ginseng root rot can be reduced, and the survival rate of ginseng plants can be improved.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The invention provides a microbial combination for reducing soil barriers, comprising Mortierella indohii, Trichoderma erinaceum, Geotrichum fragrans and Bacillus cereus.

[0007] Furthermore, the Mortierella indula is Mortierella indula CGMCC 3.12113; the Trichoderma erinaceus is Trichoderma erinaceus CGMCC 5.1548; the Geotrichum osmanthus is Geotrichum osmanthus CICC 31843; and the Bacillus cereus is Bacillus cereus CICC 21167.

[0008] The present invention also provides the use of the above-mentioned microbial combination in preparing a composite bacterial fertilizer for reducing soil barriers.

[0009] The present invention also provides a composite bacterial fertilizer for reducing soil barriers, characterized in that it comprises a fungal agent of Mortierella indica, a fungal agent of Trichoderma hedgehog, a fungal agent of Geotrichum osmanthus and a fungal agent of Bacillus cereus;

[0010] The indusia fungicide is obtained by uniformly mixing a suspension of indusia spores with corn stalk powder and then drying the mixture;

[0011] The Trichoderma argentea agent is obtained by uniformly mixing a suspension of Trichoderma argentea spores with corn stalk powder and then drying the mixture;

[0012] The osmanthus fungal agent is obtained by uniformly mixing a suspension of osmanthus fungi spores with corn stalk powder and then drying the mixture;

[0013] The Bacillus cereus agent is obtained by freeze-drying the fermented cells of Bacillus cereus;

[0014] The mass ratio of the Mortierella indula, the Trichoderma hedgehog, the Geotrichum osmanthus and the Bacillus cereus is 1:2:1:0.5.

[0015] Furthermore, the Mortierella indula is Mortierella indula CGMCC 3.12113; the Trichoderma erinaceus is Trichoderma erinaceus CGMCC 5.1548; the Geotrichum osmanthus is Geotrichum osmanthus CICC 31843; and the Bacillus cereus is Bacillus cereus CICC 21167.

[0016] Further, the concentrations of the spore suspension of Mortierella indula, the spore suspension of Trichoderma occidentalis and the spore suspension of Geotrichum osmanthus are all 10 7 CFU / mL;

[0017] The mass ratio of the Mortierella indula spore suspension, the Trichoderma hedgehog spore suspension, the Geotrichum osmanthus spore suspension and the corn straw powder is 1:1.

[0018] The present invention also provides a method for preparing the above-mentioned composite bacterial fertilizer for reducing soil barriers, comprising the following steps:

[0019] The spore suspension of Mortierella indusiae and corn stalk powder are uniformly mixed, and then dried to obtain the Mortierella indusiae fungicide;

[0020] The suspension of Trichoderma spores is mixed evenly with corn stalk powder, and then dried to obtain a Trichoderma agent;

[0021] The osmanthus fragrans spore suspension is mixed evenly with corn stalk powder, and then dried to obtain an osmanthus fragrans fungicide;

[0022] The fermented bacterial cells of Bacillus cereus are freeze-dried to obtain a Bacillus cereus agent;

[0023] The Mortierella indica agent, the Trichoderma hedgehog agent, the Geotrichum osmanthus agent and the Bacillus cereus agent are uniformly mixed in a mass ratio of 1:2:1:0.5 to obtain the composite bacterial fertilizer.

[0024] Furthermore, the Mortierella indula is Mortierella indula CGMCC 3.12113; the Trichoderma erinaceus is Trichoderma erinaceus CGMCC 5.1548; the Geotrichum osmanthus is Geotrichum osmanthus CICC 31843; and the Bacillus cereus is Bacillus cereus CICC 21167.

[0025] The present invention also provides application of the above-mentioned composite bacterial fertilizer in reducing soil continuous cropping obstacles.

[0026] The present invention also provides a method for reducing soil continuous cropping obstacles, comprising the step of applying the above-mentioned composite bacterial fertilizer to the continuous cropping soil.

[0027] The present invention discloses the following technical effects:

[0028] The present invention develops a composite bacterial fertilizer for reducing soil barriers, which is prepared by using Mortierella indica, Trichoderma hedgehog, Geotrichum osmanthus and Bacillus cereus in a mass ratio of 1:2:1:0.5. By applying the composite bacterial fertilizer provided by the present invention on the surface of ginseng continuous cropping soil, the soil conditions of ginseng continuous cropping land can be improved, ginseng growth can be promoted, the occurrence of ginseng root rot can be reduced, and the survival rate of ginseng plants can be improved. DETAILED DESCRIPTION

[0029] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0030] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0031] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0032] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0033] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0034] The strains and culture media used in the following examples are as follows:

[0035] Mortierella indohii CGMCC 3.12113 and Trichoderma erinaceum CGMCC 5.1548 were purchased from China General Microbiological Culture Collection (CGMCC); Geotrichum fragrans CICC 31843, Trichoderma harzianum CICC 13010, Bacillus cereus CICC 21167 and Bacillus subtilis CICC24713 were purchased from China Industrial Microbiological Culture Collection (CICC).

[0036] PDA liquid culture medium: 200 g / L potato and 20 g / L sucrose.

[0037] PDA solid culture medium: potato 200 g / L, sucrose 20 g / L and agar 20 g / L.

[0038] LB liquid medium: 10 g / L tryptone, 5 g / L yeast extract and 10 g / L sodium chloride.

[0039] LB solid medium: tryptone 10 g / L, yeast extract 5 g / L, sodium chloride 10 g / L and agar 20 g / L.

[0040] Solid fermentation medium: composed of the following components by mass: 80 parts of rice husk, 10 parts of corn stalk powder, 4 parts of corn flour, 4 parts of soybean meal, 1 part of MgSO4 and 100 parts of water.

[0041] Example 1 Preparation of composite bacterial fertilizer

[0042] 1. Preparation of Mortierella indica

[0043] (1) Solid slant culture: Mortierella indula CGMCC 3.12113 was inoculated into PDA solid medium and cultured at 28°C for 3 days to activate the strain.

[0044] (2) Primary seed culture: The strain activated in step (1) was inoculated into PDA liquid culture medium and cultured at 28° C. and 120 rpm for 3 days to obtain a primary seed solution.

[0045] (3) Secondary seed culture: The primary seed solution obtained in step (2) was inoculated into a 5 L Erlenmeyer flask (containing PDA liquid culture medium) at an inoculum size of 8%, and cultured at 28° C. and 120 rpm for 3 days to obtain the secondary seed solution.

[0046] (4) The secondary seed liquid was inoculated into a container containing sterile solid fermentation medium at an inoculum rate of 3%, and stirred every 48 hours to prevent the medium from caking. After culturing at 28°C for 7 days, sterile water was added and microwaved for 30 minutes to obtain a spore suspension with a concentration of 10 8 CFU / mL and above.

[0047] (5) The concentration of the spore suspension obtained in step (4) was adjusted to 10 7 CFU / mL, and evenly mixed with corn straw powder in a mass ratio of 1:1, and dried at 40°C to obtain the Mortierella indica fungicide.

[0048] 2. Preparation of Trichoderma serrata

[0049] (1) Solid slant culture: Trichoderma erinaceus CGMCC 5.1548 was inoculated into PDA solid culture medium and cultured at 28°C for 3 days to activate the strain.

[0050] (2) Primary seed culture: The strain activated in step (1) was inoculated into PDA liquid culture medium and cultured at 28° C. and 120 rpm for 3 days to obtain a primary seed solution.

[0051] (3) Secondary seed culture: The primary seed solution obtained in step (2) was inoculated into a 5 L Erlenmeyer flask (containing PDA liquid culture medium) at an inoculum size of 8%, and cultured at 28° C. and 120 rpm for 3 days to obtain the secondary seed solution.

[0052] (4) The secondary seed liquid was inoculated into a container containing sterile solid fermentation medium at an inoculum rate of 3%, and stirred every 48 hours to prevent the medium from caking. After culturing at 28°C for 7 days, sterile water was added and microwaved for 30 minutes to obtain a spore suspension with a concentration of 10 8 CFU / mL and above.

[0053] (5) The concentration of the spore suspension obtained in step (4) was adjusted to 10 7 CFU / mL, mixed evenly with corn straw powder in a mass ratio of 1:1, and dried at 40°C to obtain the hedgehog Trichoderma agent.

[0054] 3. Preparation of Osmanthus fragrans fungicide

[0055] (1) Solid slant culture: Geotrichum osmanthus CICC 31843 was inoculated into PDA solid medium and cultured at 28°C for 3 days to activate the strain.

[0056] (2) Primary seed culture: The strain activated in step (1) was inoculated into PDA liquid culture medium and cultured at 28° C. and 120 rpm for 3 days to obtain a primary seed solution.

[0057] (3) Secondary seed culture: The primary seed solution obtained in step (2) was inoculated into a 5 L Erlenmeyer flask (containing PDA liquid culture medium) at an inoculum size of 8%, and cultured at 28° C. and 120 rpm for 3 days to obtain the secondary seed solution.

[0058] (4) The secondary seed liquid was inoculated into a container containing sterile solid fermentation medium at an inoculum rate of 3%, and stirred every 48 hours to prevent the medium from caking. After culturing at 28°C for 7 days, sterile water was added and microwaved for 30 minutes to obtain a spore suspension with a concentration of 10 8 CFU / mL and above.

[0059] (5) The concentration of the spore suspension obtained in step (4) was adjusted to 10 7 CFU / mL, mixed evenly with corn straw powder in a mass ratio of 1:1, and dried at 40°C to obtain the osmanthus fungicide.

[0060] 4. Preparation of Bacillus cereus inoculant

[0061] (1) Solid slant culture: Bacillus cereus CICC 21167 was inoculated into LB solid medium and cultured at 37°C for 1 day to activate the strain.

[0062] (2) Primary seed culture: The strain activated in step (1) was inoculated into LB liquid culture medium and cultured at 37° C. and 160 rpm for 1 day to obtain a primary seed solution.

[0063] (3) Secondary seed culture: The primary seed solution obtained in step (2) was inoculated into a 5 L Erlenmeyer flask (containing LB liquid culture medium) at an inoculum size of 8%, and cultured at 37° C., 160 rpm for 1 day to obtain the secondary seed solution.

[0064] (4) The secondary seed liquid obtained in step (3) is inoculated into a fermentation medium (i.e., LB liquid medium) at an inoculum rate of 10%, and fermented at 37° C. and 180 rpm for 2 days, and then the fermentation liquid is centrifuged to obtain bacterial cells, which are freeze-dried to obtain a Bacillus cereus inoculum.

[0065] 5. Preparation of compound fertilizer

[0066] The fungal agent of Mortierella indole, the fungal agent of Trichoderma hedgehog, the fungal agent of Geotrichum osmanthus and the fungal agent of Bacillus cereus were uniformly mixed in a mass ratio of 1:2:1:0.5 to obtain a composite bacterial fertilizer.

[0067] Comparative Example 1

[0068] The same as Example 1, except that the Mortierella indoleica agent, the Trichoderma hedgehog agent, the Geotrichum osmanthus agent and the Bacillus cereus agent are uniformly mixed in a mass ratio of 1:1:1:0.5 to obtain a composite bacterial fertilizer.

[0069] Comparative Example 2

[0070] Same as Example 1, except that Trichoderma harzianum CGMCC 5.1548 was replaced by Trichoderma harzianum CICC 13010.

[0071] Comparative Example 3

[0072] Same as Example 1, except that Bacillus cereus CICC 21167 is replaced by Bacillus subtilis CICC24713.

[0073] Example 2

[0074] 1. Experimental Materials

[0075] The ginseng used in the planting experiment was 2-year-old ginseng seedlings; the planting experiment was carried out in a field where ginseng had been planted for 5 consecutive years.

[0076] 2. Experimental Methods

[0077] After deep plowing of 30 cm in the ginseng planting field, the field was randomly divided into 5 experimental areas, and the composite bacterial fertilizer prepared in Example 1 and Comparative Examples 1-3 was evenly spread on the soil surface at an application rate of 1.5 kg / mu, and then 2-year-old ginseng seedlings were planted (the planting density in each area was the same), and then conventional field management was carried out according to local management methods; at the same time, plants without the addition of composite bacterial fertilizer were used as controls. After one year of planting, the ginseng yield, root weight per plant, and survival rate of each experimental area were counted.

[0078] 3. Experimental results

[0079] One year after planting, the statistical results of the number of plants infected with root rot, the root weight of a single plant and the survival rate in each experimental area are shown in Table 1. As can be seen from Table 1, the composite bacterial fertilizer designed by the present invention, prepared by using the indusia fungicide, the hedgehog fungicide, the osmanthus fungicide and the Bacillus cereus fungicide in a mass ratio of 1:2:1:0.5, can effectively increase the single root fresh weight and survival rate of ginseng plants, and reduce the incidence of root rot, thereby effectively reducing soil continuous cropping obstacles.

[0080] Table 1 Number of plants infected with root rot, root weight per plant and survival rate in each experimental area

[0081] Experimental Area Fresh weight of single root (g) Survival rate (g) Root rot incidence (%) Control group 9.20 86.4 16.7 Example 1 <![CDATA[11.38 * ]]> <![CDATA[95.7 ** ]]> <![CDATA[6.2 ** ]]> Comparative Example 1 9.35 <![CDATA[90.4 * ]]> <![CDATA[9.3 * ]]> Comparative Example 2 9.87 88.7 13.9 Comparative Example 3 9.31 87.5 <![CDATA[9.2 * ]]>

[0082] Note: Compared with the control group, * P<0.05, ** P<0.01.

[0083] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A microbial combination for reducing soil barriers, characterized in that: These include Mortierella indohii, Trichoderma erinaceum, Geotrichum fragrans and Bacillus cereus.

2. The microbial combination according to claim 1, characterized in that The Mortierella indula is Mortierella indula CGMCC 3.12113; the Trichoderma occidentalis is Trichoderma occidentalis CGMCC 5.1548; the Geotrichum osmanthus is Geotrichum osmanthus CICC31843; and the Bacillus cereus is Bacillus cereus CICC 21167.

3. Use of the microbial combination as claimed in claim 1 or 2 in the preparation of a compound bacterial fertilizer for reducing soil barriers.

4. A composite bacterial fertilizer for reducing soil barriers, characterized in that: Including Mortierella indula, Trichoderma hedgehog, Geotrichum osmanthus and Bacillus cereus; The indusia fungicide is obtained by uniformly mixing a suspension of indusia spores with corn stalk powder and then drying the mixture; The Trichoderma argentea agent is obtained by uniformly mixing a suspension of Trichoderma argentea spores with corn stalk powder and then drying the mixture; The osmanthus fungal agent is obtained by uniformly mixing a suspension of osmanthus fungi spores with corn stalk powder and then drying the mixture; The Bacillus cereus agent is obtained by freeze-drying the fermented cells of Bacillus cereus; The mass ratio of the Mortierella indula, the Trichoderma hedgehog, the Geotrichum osmanthus and the Bacillus cereus is 1:2:1:0.

5.

5. The composite bacterial fertilizer according to claim 4, characterized in that: The Mortierella indula is Mortierella indula CGMCC3.12113; the Trichoderma erinaceus is Trichoderma erinaceus CGMCC 5.1548; the Geotrichum osmanthus is Geotrichum osmanthus CICC 31843; and the Bacillus cereus is Bacillus cereus CICC 21167.

6. The composite bacterial fertilizer according to claim 4, characterized in that: The concentrations of the spore suspension of Mortierella indula, the spore suspension of Trichoderma occidentalis and the spore suspension of Geotrichum osmanthus were all 10 7 CFU / mL; The mass ratio of the Mortierella indula spore suspension, the Trichoderma hedgehog spore suspension, the Geotrichum osmanthus spore suspension and the corn straw powder is 1:

1.

7. A method for preparing a composite bacterial fertilizer for reducing soil barriers as claimed in any one of claims 4 to 6, characterized in that: The following steps are involved: The spore suspension of Mortierella indusiae and corn stalk powder are uniformly mixed, and then dried to obtain the Mortierella indusiae fungicide; The suspension of Trichoderma spores is mixed evenly with corn stalk powder, and then dried to obtain a Trichoderma agent; The osmanthus fragrans spore suspension is mixed evenly with corn stalk powder, and then dried to obtain an osmanthus fragrans fungicide; The fermented bacterial cells of Bacillus cereus are freeze-dried to obtain a Bacillus cereus agent; The Mortierella indica agent, the Trichoderma hedgehog agent, the Geotrichum osmanthus agent and the Bacillus cereus agent are uniformly mixed in a mass ratio of 1:2:1:0.5 to obtain the composite bacterial fertilizer.

8. The preparation method according to claim 7, characterized in that: The Mortierella indula is Mortierella indula CGMCC3.12113; the Trichoderma erinaceus is Trichoderma erinaceus CGMCC 5.1548; the Geotrichum osmanthus is Geotrichum osmanthus CICC 31843; and the Bacillus cereus is Bacillus cereus CICC 21167.

9. Use of the composite bacterial fertilizer as claimed in any one of claims 4 to 6 in reducing soil continuous cropping obstacles.

10. A method for reducing soil continuous cropping obstacles, characterized in that: The method comprises the step of applying the compound bacterial fertilizer according to any one of claims 4 to 6 to the continuously cropped soil.