Method for promoting soybean growth by inducing bacteriophage

Abstract

The invention belongs to the technical field of agricultural products, and particularly relates to a method for promoting soybean growth by inducing bacteriophages. The method specifically comprises the step of applying a phage inducer as an accelerant in a soil environment for planting the soybeans so as to promote the growth of the soybeans. According to the invention, phage induced release is utilized, soil phage abundance is increased, soil nutrient condition is improved, and root growth is promoted in combination with chemical fertilizer addition. In addition, due to the wide existence of the bacteriophage, the resistance ability of soybeans to pathogens and other environmental stress is enhanced, and soil health and agricultural sustainable development are facilitated.

Description

Technical Field

[0001] This invention belongs to the field of agricultural product technology, specifically relating to a method for promoting soybean growth by inducing bacteriophages. Background Technology

[0002] Excessive use of pesticides and irrational farming practices in agricultural production significantly impact soil health and productivity. Maintaining soil health is one of the most critical issues for sustainable agricultural development. While biological methods such as applying growth-promoting bacteria and probiotics are increasingly used for soil health maintenance and plant growth enhancement, they often compete with native microbial communities after application, hindering microbial colonization and hindering their effectiveness. Therefore, their widespread application still faces certain challenges.

[0003] Soil bacteriophages, due to their regulatory role in microbial communities, have emerged as a novel tool for bioremediation, with technologies gradually being developed for controlling soil-borne diseases. Furthermore, the role of bacteriophages in maintaining soil health and promoting nutrient cycling is increasingly recognized; their lytic action against soil bacteria effectively accelerates the turnover of soil nutrients. However, due to the specificity of bacteriophages in infecting their host, there are currently no effective technologies for developing and applying them in this area. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the prior art and provide a method for promoting soybean growth by inducing bacteriophages.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] A method for promoting soybean growth by inducing bacteriophages involves applying a bacteriophage inducer as a promoter in the soil environment where soybeans are grown, thereby promoting soybean growth.

[0007] The phage inducer is one or more of mitomycin C and / or N-acylhomoserine lactone.

[0008] Among them, N-acyl homoserine lactones include one or more of N-butyroyl, N-hexanoyl, N-heptanoyl, N-octanoyl, N-3-oxo-hexanoyl, N-3-oxo-octanoyl, and N-3-oxo-decanoyl-L-homoserine lactones.

[0009] The phage inducer is mixed with fertilizer and applied to the soil for planting soybeans at the time of sowing.

[0010] The amount of the phage inducer added is 0.5‰-1‰ of the fertilizer mass; after the phage inducer is mixed with the fertilizer, the amount applied per acre when the mixture is applied to the soil is 50 kg.

[0011] The fertilizer is an N-P2O5-K2O compound fertilizer with a formula ratio of 25-18-15.

[0012] Application of a bacteriophage inducer as a soybean growth promoter.

[0013] The phage inducer is one or more of mitomycin C and / or N-acylhomoserine lactone.

[0014] As can be seen from the above description of the present invention, compared with the prior art, the beneficial effects of the present invention are:

[0015] This invention utilizes chemical inducers to release bacteriophages, which in turn promote the activity of soybean rhizosphere microorganisms and nutrient turnover, enhance rhizosphere nitrogen fixation, and further promote soybean rhizosphere development. Furthermore, the presence of bacteriophages can potentially regulate the microbial community structure, inhibit the spread of soil-borne pathogens, and enhance stress resistance. Attached Figure Description

[0016] Figure 1 This is a comparison chart showing the number of soil viruses with and without the addition of an inducer, provided in an embodiment of the present invention.

[0017] Figure 2 This is a comparison diagram of soybean root growth with and without the addition of an inducing agent, provided in an embodiment of the present invention.

[0018] Figure 3 This is a comparison diagram of soil nutrient status with and without the addition of an inducing agent, provided in an embodiment of the present invention. Detailed Implementation

[0019] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0020] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0021] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0022] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This application specification and embodiments are merely exemplary.

[0023] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0024] All raw materials used in the following embodiments of the present invention are commercially available.

[0025] This invention utilizes chemical induction technology to accelerate the release of mild bacteriophages from the microbial host, increasing phage abundance in the soil and promoting soybean growth by improving soil nutrient turnover. More specifically, this invention uses chemical inducers to induce phage release, enhancing rhizosphere nitrogen fixation by promoting soybean rhizosphere microbial activity and nutrient turnover, thereby further promoting soybean rhizosphere development. Furthermore, the presence of phages can potentially regulate microbial community structure, inhibit the spread of soil-borne pathogens, and enhance stress resistance. The chemical inducers of this invention indirectly promote fertilizer fertility and soybean growth, while also maintaining soil health, thus better supporting the sustainable development of black soil.

[0026] In the following examples, the soybean seeds were Williams 82; the fertilizer was an N-P2O5-K2O compound fertilizer with a formula ratio of 25-18-15. The fertilizer in the examples contained urea (N content 46%), superphosphate (P2O5 content 12%), and potassium chloride (K2O content 60%).

[0027] Example 1

[0028] An indoor pot experiment was conducted using Northeast China black soil. 800g of soil was added to each pot, along with three surface-sterilized soybean seeds buried in the soil. A mixture of fertilizer and mitomycin C was then added. The soybean pots were cultured at room temperature for 7 days, with a light-dark cycle of 16 hours of light and 8 hours of darkness. The abundance of bacteriophages in the root zone soil was determined using fluorescence microscopy at different culture times. A control group without bacteriophage inducers was used (see [link to experiment]). Figure 1 , Figure 2 and Figure 3 ).

[0029] The final concentration of mitomycin C in the above mixture of fertilizer and mitomycin C was 0.5 μg / g dry soil. The fertilizer contained urea (46% N content), superphosphate (12% P2O5 content), and potassium chloride (60% K2O content).

[0030] Example 2

[0031] An indoor pot experiment was conducted using Northeast China black soil. 800g of soil was added to each pot, along with three surface-sterilized soybean seeds buried in the soil. A mixture of fertilizer and the phage inducer mitomycin C was then added. The soybean pots were cultured at room temperature for 21 days, with a photoperiod alternating between 16 hours of light and 8 hours of darkness. The abundance of phages in the root zone soil was determined using fluorescence microscopy at different culture times. A control group without the phage inducer was used (see [link to experiment]). Figure 1 , Figure 2 and Figure 3 ).

[0032] The final concentration of mitomycin C in the above mixture of fertilizer and mitomycin C was 0.5 μg / g dry soil. The fertilizer contained urea (46% N content), superphosphate (12% P2O5 content), and potassium chloride (60% K2O content).

[0033] Example 3

[0034] An indoor pot experiment was conducted using Northeast China black soil. 800g of soil was added to each pot, along with three surface-sterilized soybean seeds buried in the soil. A mixture of fertilizer and the phage inducer mitomycin C was then added. The soybean pots were cultured at room temperature for 28 days, with a photoperiod alternating between 16 hours of light and 8 hours of darkness. The abundance of phages in the root zone soil was determined using fluorescence microscopy at different culture times. A control group without the phage inducer was used (see [link to experiment]). Figure 1 , Figure 2 and Figure 3 ).

[0035] The final concentration of mitomycin C in the above mixture of fertilizer and mitomycin C was 0.5 μg / g dry soil. The fertilizer contained urea (46% N content), superphosphate (12% P2O5 content), and potassium chloride (60% K2O content).

[0036] In the above embodiments, the phage abundance in the root soil was obtained by fluorescence microscopy counting method, referring to the virus abundance calculation method in Liang et al. (2020) and patent CN 115927542 A. Figure 1 As can be seen, the abundance of bacteriophages in the treatment with the addition of mitomycin C in each embodiment increased significantly by 1-2 times compared with that without the addition, indicating that the bacteriophages were successfully induced in each embodiment.

[0037] Simultaneously, the root growth of soybeans was compared at different growth stages in each embodiment, and the root growth and development were as follows: Figure 2 This shows that the addition of mitomycin C significantly promoted root development.

[0038] Furthermore, soil nutrient conditions such as Figure 3 In each embodiment, the addition of mitomycin C at different soybean growth stages significantly increased the content of nitrate nitrogen, ammonium nitrogen, and dissolved organic carbon in the soil.

[0039] Example 4

[0040] An indoor pot experiment was conducted using Northeast China black soil. 800g of soil was added to each pot, along with three surface-sterilized soybean seeds buried in the soil. Fertilizer and the phage inducer N-hexanoyl-L-homoserine lactone were then added. The soybean pots were cultured at room temperature for 28 days, with a light cycle of 16 hours of light and 8 hours of darkness. A control group without the phage inducer was used.

[0041] The final concentration of N-hexanoyl-L-homoserine lactone in the mixture of the above-mentioned fertilizer and N-hexanoyl-L-homoserine lactone is 0.5 μg / g dry soil. The fertilizer contains urea (N content 46%), superphosphate (P2O5 content 12%), and potassium chloride (K2O content 60%).

[0042] In the treatment with N-hexanoyl-L-homoserine lactone, the phage abundance increased significantly by 2 times compared with that without addition. Soybean root length and dry weight increased by 1.5 times and 1.2 times, respectively, compared with that without addition.

[0043] References:

[0044] Liang X,Wagner RE,Li B,Zhang N and Radosevich M(2020)Quorum SensingSignals Alter in vitro Soil Virus Abundance and Bacterial CommunityComposition.Front.Microbiol.11:1287.doi:10.3389 / fmicb.2020.01287

[0045] The above description is merely a preferred embodiment of the present invention and should not be construed as limiting the scope of the present invention. All equivalent changes and modifications made in accordance with the scope of the patent application and the contents of the specification of the present invention should still fall within the scope of the patent of the present invention.

Claims

1. A method for promoting soybean growth by inducing bacteriophages, characterized in that: Applying bacteriophage inducers as promoters to the soil environment in which soybeans are grown can promote soybean growth.

2. The method for promoting soybean growth by inducing bacteriophages according to claim 1, characterized in that: The phage inducer is one or more of mitomycin C and / or N-acylhomoserine lactone.

3. The method for promoting soybean growth by inducing bacteriophages according to claim 1 or 2, characterized in that: The phage inducer is mixed with fertilizer and applied to the soil for planting soybeans at the time of sowing.

4. The method for promoting soybean growth by inducing bacteriophages according to claim 3, characterized in that: The amount of the phage inducer added is 0.5‰-1‰ of the fertilizer mass; after the phage inducer is mixed with the fertilizer, the amount applied per acre when the mixture is applied to the soil is 50 kg.

5. The method for promoting soybean growth by inducing bacteriophages according to claim 3, characterized in that: The fertilizer is an N-P2O5-K2O compound fertilizer with a formula ratio of 25-18-15.

6. The application of a phage inducer as a soybean growth promoter.

7. The application according to claim 6, characterized in that: The phage inducer is one or more of mitomycin C and / or N-acylhomoserine lactone.

Images