Microbial agent for promoting growth of Chinese wolfberry fruits and application of microbial agent

Abstract

The invention discloses a microbial agent for promoting growth of Chinese wolfberry fruits and application of the microbial agent, and belongs to the technical field of agricultural biology. The invention provides a microbial agent, which is prepared from streptomyces albidoflavus, bacillus mucilaginosus, bacillus velezensis, bacillus mucilaginosus and bacillus velezensis, and the microbial agent is prepared from streptomyces albidoflavus, bacillus mucilaginosus, bacillus mucilaginosus, bacillus velezensis, bacillus mucilaginosus, bacillus mucilaginosus, bacillus mucilaginosus, bacillus mucilaginosus, bacillus mucilaginosus, bacillus mucilaginosus, bacillus mucilaginosus, bacillus mucilaginosus, bacillus mucilaginosus, bacillus mucilaginosus and bacillus subtilis. The streptomyces albidoflavus, the bacillus mucilaginosus and the bacillus velezensis are inoculated into a fermentation culture medium in a seed liquid form; the inoculum size volume ratio of the seed solution of the streptomyces albidoflavus to the seed solution of the bacillus mucilaginosus to the seed solution of the bacillus velezensis is 4: 3: 2. The microbial agent disclosed by the invention provides an innovative biological solution for agricultural sustainable development, and is beneficial to reducing the use of chemical inputs and promoting the development of ecological agriculture.

Description

Technical Field

[0001] The invention relates to the field of agricultural biotechnology, in particular to a microbial agent for promoting wolfberry growth and application thereof. Background Art

[0002] In modern agricultural production, wolfberry (Lycium barbarum L.) is an important economic crop and is widely planted around the world for its rich nutritional value and medicinal value. However, the wolfberry planting industry is facing multiple challenges, including soil degradation, frequent pests and diseases, and soil microecological imbalance caused by unreasonable fertilization. These problems not only affect the yield and quality of wolfberry, but also threaten the sustainability of the ecological environment.

[0003] Traditional agricultural management methods, such as the large-scale application of chemical fertilizers and pesticides, can increase production in the short term, but have serious negative effects in the long run. Excessive use of chemical fertilizers can easily lead to soil salinization and acidification, destroy the balance of soil microorganisms, reduce soil fertility, and may also cause water pollution. Although frequent application of pesticides can temporarily control pests and diseases, it promotes the enhancement of pest resistance, reduces natural enemies, destroys the natural balance of the ecosystem, and increases food safety risks.

[0004] In recent years, with the rapid development of biotechnology, microbial agents have been increasingly valued as a green and environmentally friendly auxiliary means of agricultural production. Microbial agents use beneficial microorganisms in nature to improve soil physical and chemical properties, inhibit pathogens, promote crop growth, and enhance crop resistance. However, there are still few microbial agents specially developed for the growth characteristics and planting environment of wolfberry on the market, and their efficacy varies, making it difficult to meet the comprehensive needs of high-yield, high-quality and ecological safety of wolfberry. Therefore, it is particularly important to develop a safe, environmentally friendly microbial agent that can effectively promote the growth of wolfberry. Summary of the invention

[0005] The purpose of the present invention is to provide a microbial agent for promoting the growth of wolfberry and its application, so as to solve the problems existing in the above-mentioned prior art. The microbial agent of the present invention can not only promote the growth of wolfberry, but also increase the yield of wolfberry.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] Technical solution 1: A microbial agent, including Streptomyces albidoflavus, Bacillus mucilaginosus Krassilnikov and Bacillus subtilis.

[0008] Furthermore, the bacterial content of the seed liquid of Streptomyces albicans, Bacillus colloids and Bacillus velezii was 1.0×10 8 CFU / mL.

[0009] Furthermore, the Streptomyces albus, Bacillus subtilis and Bacillus velezii are inoculated into the fermentation medium in the form of biological seed liquid; the inoculation volume ratio between the seed liquids of the Streptomyces albus, Bacillus subtilis and Bacillus velezii is 4:3:2.

[0010] Technical Solution 2: Application of the microbial agent in promoting crop growth.

[0011] Furthermore, the crops include wolfberry.

[0012] Technical solution three: Application of the microbial agent in increasing crop yield.

[0013] Furthermore, the crops include wolfberry.

[0014] The present invention discloses the following technical effects:

[0015] The present invention provides a microbial agent that combines three beneficial microorganisms with unique biological activities - Streptomyces albus, Bacillus colloids and Bacillus velez. In the specific preparation process, the three strains are cultivated to a high density state to form their own seed liquid to ensure that the number of colony forming units (CFU) contained in each milliliter of seed liquid is sufficient. Based on the study of the functional characteristics of each strain and the interaction between them, the present invention mixes and inoculates the seed liquids of the three strains in a ratio of 4:3:2, which can maximize the overall biological effect.

[0016] The present invention promotes plant growth and increases crop yields through biotechnology, provides an innovative biological solution for sustainable agricultural development, helps reduce the use of chemical inputs, and promotes the development of ecological agriculture. In short, the microbial agent of the present invention can not only promote the healthy growth of plants, increase crop yield and quality, but also reduce dependence on pesticides, which is of great significance to environmental protection and ecological balance. DETAILED DESCRIPTION

[0017] 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.

[0018] 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.

[0019] 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.

[0020] 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.

[0021] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0022] Streptomyces albidoflavus product number BMZ099271, Bacillus mucilaginosus Krassilnikov product number BMZ102180, Bacillus subtilis product number BMZ135815 and Bacillus velezensis product number B64646 were purchased from Ningbo Mingzhou Biotechnology Co., Ltd.

[0023] The purity of sorbitol, Tween-80, methylparaben and sodium carboxymethyl cellulose must be industrial purity or above.

[0024] Example 1

[0025] A liquid inoculant containing microbial inoculants

[0026] The preparation method of liquid bacterial agent is as follows: the strains of Streptomyces albicans, Bacillus colloids and Bacillus velez are activated on LB plates and then inoculated into LB culture solution for liquid fermentation. The concentration of the liquid bacterial agent is determined to be 1.0×10 8 CFU / mL, the micro-white yellow Streptomyces seed liquid, the gelatinous Bacillus seed liquid and the Velez Bacillus seed liquid were mixed in a volume ratio of 4:3:2, and the seed liquid was formed and continued to ferment. After the fermentation was completed, 0.5% of the sterilized UV protective agent sorbitol, 1% of the sterilized surfactant Tween-80, 0.5% of the sterilized preservative methyl paraben and 1% of the sterilized stabilizer sodium carboxymethyl cellulose were added according to the volume ratio of the bacterial liquid to prepare a liquid bacterial agent. After the mixture was stabilized for 48 hours, the concentration was measured to reach the technical indicators of agricultural microbial inoculants in the national standard GB20287-2006, forming a liquid bacterial agent 1.

[0027] Comparative Example 1

[0028] A liquid inoculant containing microbial inoculants

[0029] The preparation method of liquid bacterial agent is as follows: the strains of Streptomyces albicans, Bacillus colloids and Bacillus velez are activated on LB plates and then inoculated into LB culture solution for liquid fermentation. The concentration of the liquid bacterial agent is determined to be 1.0×10 8 CFU / mL, the micro-white yellow Streptomyces seed liquid, the gelatinous Bacillus seed liquid and the Velez Bacillus seed liquid were mixed in a volume ratio of 1:1:1, and the seed liquid was formed and continued to ferment. After the fermentation was completed, 0.5% of the sterilized UV protective agent sorbitol, 1% of the sterilized surfactant Tween-80, 0.5% of the sterilized preservative methyl paraben and 1% of the sterilized stabilizer sodium carboxymethyl cellulose were added according to the volume ratio of the bacterial liquid to prepare a liquid bacterial agent. After the mixture was stabilized for 48 hours, the concentration was measured to reach the technical indicators of agricultural microbial inoculants in the national standard GB20287-2006, forming a liquid bacterial agent 2.

[0030] Comparative Example 2

[0031] A liquid inoculant containing microbial inoculants

[0032] The preparation method of liquid bacterial agent is as follows: after activating Bacillus mucilaginosus and Bacillus velez on LB plate, inoculate into LB culture solution for liquid fermentation, and determine the concentration of the liquid bacterial agent to reach 1.0×10 8CFU / mL, the micro-white yellow Streptomyces seed liquid and the gelatinous Bacillus seed liquid were mixed in a volume ratio of 4:3, and the seed liquid was formed and continued to ferment. After the fermentation was completed, 0.5% sterilized UV protective agent sorbitol, 1% sterilized surfactant Tween-80, 0.5% sterilized preservative methyl paraben and 1% sterilized stabilizer sodium carboxymethyl cellulose were added according to the volume ratio of the bacterial liquid to prepare a liquid bacterial agent. After mixing and stabilizing for 48 hours, the concentration was measured to meet the technical indicators of agricultural microbial inoculants in the national standard GB20287-2006, forming a liquid bacterial agent 3.

[0033] Comparative Example 3

[0034] A liquid inoculant containing microbial inoculants

[0035] The preparation method of liquid microbial agent is as follows: the strains of Streptomyces albicans, Bacillus subtilis and Bacillus velez are activated on LB plates and then inoculated into LB culture solution for liquid fermentation. The concentration of the liquid microbial agent is determined to be 1.0×10 8 CFU / mL, the micro-white yellow Streptomyces seed liquid, the gelatinous Bacillus seed liquid and the Velez Bacillus seed liquid were mixed in a volume ratio of 4:3:2, and the seed liquid was formed and continued to ferment. After the fermentation was completed, 0.5% of the sterilized UV protective agent sorbitol, 1% of the sterilized surfactant Tween-80, 0.5% of the sterilized preservative methyl paraben and 1% of the sterilized stabilizer sodium carboxymethyl cellulose were added according to the volume ratio of the bacterial liquid to prepare a liquid bacterial agent. After the mixture was stabilized for 48 hours, the concentration was measured to reach the technical indicators of agricultural microbial inoculants in the national standard GB20287-2006, forming a liquid bacterial agent 4.

[0036] Experimental example

[0037] Field test report on liquid microbial agent 1-4 for promoting the growth of wolfberry

[0038] 1. Experimental crops: wolfberry, variety Ningqi No. 1.

[0039] 2. Environmental conditions

[0040] The experiment was set up in Hantianling Village, Hexi Town, Tongxin County, Wuzhong City, Ningxia Hui Autonomous Region. The terrain is flat and the fertility is medium. The wolfberry trees are 8 years old and the row spacing is 1.5 meters × 2.0 meters. The experiment was carried out in 2022. During the experiment, watering was carried out three times on May 6, June 8 and July 11, and other field management was carried out as usual.

[0041] 3. Experimental Design and Arrangement

[0042] 3.1 Bacterial Agents

[0043] 3.1.1 Test microbial agents

[0044] Liquid bacterial agents 1-4 prepared in Example 1 and Comparative Examples 1-3.

[0045] 3.1.2 Dosage of microbial agent

[0046] 500kg / mu, irrigate roots once.

[0047] 3.2 Experimental arrangements

[0048] The experiment adopted a large-scale plot design with an area of ​​1.0 mu and 30 plants in each treatment group.

[0049] 3.3 Sterilization method

[0050] 3.3.1 Usage

[0051] In spring, before wolfberry sprouts, root irrigation should be carried out on April 22. Select 3 points around each tree, about 50 cm away from the base of the stem, insert a topdressing gun into the soil, pause for 5 seconds at each point, and pour the liquid medicine into the roots of the wolfberry.

[0052] 3.3.2 Sterilization equipment

[0053] Tractor, bucket (1500kg), fertilizer gun.

[0054] 3.3.3 Sterilization time and frequency

[0055] Root irrigation was carried out on April 17, and the bacteria were applied once.

[0056] 3.4 Usage capacity

[0057] According to the experimental design, before wolfberry sprouts, the experimental bacterial agent is prepared into a bacterial solution according to the designed dosage, and the roots are irrigated at 500kg / mu (4-5L / plant).

[0058] 4 Investigation, recording and measurement methods

[0059] 4.1 Survey time and frequency

[0060] Record the crown and stem diameter of the wolfberry tree when irrigating the roots; measure and record the yield at several times (6-7 times) when picking wolfberries; record the crown and stem diameter again after the wolfberry leaves fall.

[0061] 4.2 Survey Methods

[0062] 4.2.1 Investigation of tree crown, stem diameter and yield

[0063] Avoid the edge rows and select the middle rows according to the characteristics of the plot. In each treatment, 1-3 rows were surveyed according to the row length, with an interval of 1-2 rows.

[0064] 4.3 Direct impact on crops

[0065] According to irregular field observations, plants in each treatment area of ​​the test microbial agent grew normally without any adverse effects, indicating that the test microbial agent is safe for wolfberry within the test dosage range.

[0066] 4.4 Impact on crop yield

[0067] After wolfberry matured, one row was selected for each treatment (two rows were selected for short plots) to record the number of plants. Five harvests were carried out on wolfberry in the experimental field according to the number of harvests. The fresh weight yield was recorded for each harvest, and the total yield was calculated by merging them. The yield per plant was then averaged.

[0068] 5 Results and Analysis

[0069] The effects of liquid microbial agents 1-3 on the growth of wolfberry are shown in Table 1. The blank control was root irrigation without any microbial agent.

[0070] Table 1

[0071]

[0072] The data in Table 1 show that the stem diameter of wolfberry trees has almost no change between the beginning and the end of the year, and the application of microbial agent has no difference in effect on 8-year-old wolfberry trees. However, the crown width has changed greatly, and the application of liquid microbial agent 1 in Example 1 has the greatest increase in the crown width of wolfberry trees.

[0073] Table 2 shows the effects of the bacterial agents in each treatment group on wolfberry yield.

[0074] Table 2

[0075]

[0076] The results in Table 2 show that the application of the liquid bacterial agent prepared in Example 1 can increase the yield of wolfberry, with a single plant yield of 8.93 kg. Compared with the single plant yield of the liquid bacterial agent prepared in Comparative Examples 1-3, the maximum increase per plant reached 2.69 kg.

[0077] 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 agent, characterized in that: These include Streptomyces albidoflavus, Bacillus mucilaginosus Krassilnikov, and Bacillus subtilis.

2. The microbial agent according to claim 1, characterized in that The bacterial content of the seed liquid of Streptomyces albicans, Bacillus colloids and Bacillus velezii was 1.0×10 8 CFU / mL.

3. The microbial agent according to claim 2, characterized in that: The Streptomyces albus, Bacillus colloids and Bacillus velezii are inoculated into a fermentation medium in the form of seed liquid; the inoculation volume ratio between the seed liquids of the Streptomyces albus, Bacillus colloids and Bacillus velezii is 4:3:

2.

4. Use of the microbial agent according to any one of claims 1 to 3 in promoting crop growth.

5. The use according to claim 4, characterized in that: The crops include wolfberry.

6. Use of the microbial agent according to any one of claims 1 to 3 in increasing crop yield.

7. The use according to claim 6, characterized in that: The crops include wolfberry.

Images