A microbial formulation for improving soil and increasing productivity and its liquid mulch application

Abstract

The present disclosure relates to a microbial preparation for improving soil and increasing productivity and a liquid mulching application thereof, which contains Rhizopus arrhizus fermentation, Bacillus pumilus fermentation and carboxymethyl cellulose; the taxonomic name of the Rhizopus arrhizus is Rhizopus arrhizus, and the preservation number is CGMCC No. 40610; the taxonomic name of the Bacillus pumilus is Bacillus pumilus, and the preservation number is CGMCC No. 13208. The microbial preparation is convenient to use, green and pollution-free, and can increase the biological yield of crops, improve the water content in the soil and improve the population structure of microorganisms in the soil, and promote the expansion of the ecological niche of probiotics.

Description

Technical Field

[0001] This disclosure relates to the field of agricultural production technology and the field of soil environmental protection technology. Specifically, it relates to a microbial preparation for improving soil and increasing productivity, and its liquid mulch application. Background Technology

[0002] China has the world's largest use and coverage area of ​​plastic film mulching. The first stage of China's plastic film development, from 1979 to 1984, was primarily an introduction and experimentation phase, focusing on the import and imitation of plastic film products. The second stage, from 1985 to 1992, was mainly a technology improvement phase, including the development of planting models and mulching machinery. From 1993 onwards, the next 20 years can be considered the third stage: the technology application stage. During this stage, plastic film mulching technology developed rapidly in China. In recent years, the area covered by plastic film for crops has reached nearly 300 million mu (approximately 20 million hectares), using 75% of the world's plastic film. In many regions, plastic film mulching has changed agricultural planting patterns, increasing crop water use efficiency and yield by about 30%, making a significant contribution to ensuring the safe supply of agricultural products in my country. However, the use of plastic film is not without its drawbacks. While increasing yields and efficiency and ensuring the production of grains, vegetables, and cash crops, the side effects of large-scale use of plastic film have begun to emerge: plastic film pollution. The raw material for plastic film is polyethylene, which degrades very slowly in soil, potentially taking a century to degrade. The long-term and large-scale application of plastic film, coupled with the difficulty of recycling, has led to an increasingly serious problem of residual plastic film pollution in my country's farmland.

[0003] In July 2020, the Ministry of Agriculture and Rural Affairs issued the "Administrative Measures for Agricultural Film," requiring that "users of agricultural film should collect non-fully biodegradable agricultural film waste from the fields before its expiration date and hand it over to recycling points or recycling workers, and should not arbitrarily discard, bury, or burn it." However, the recycling of agricultural film faces many difficulties, such as lack of efficiency and inadequate recycling channels. In recent years, my country has been conducting research and development and promotion of biodegradable mulch films. Biodegradable mulch films are one of the effective measures and means to replace traditional mulch films and solve the problem of residual pollution from mulch films. Patent application number 202011573994.8 discloses a semi-coke liquid mulch film and its preparation method; patent application number 201410720959.2 discloses a liquid mulch film and its preparation method; patent application number 202011363902.3 discloses a bentonite liquid mulch film and its preparation method and application; and patent application number 201710159176.5 discloses a production method for straw composite liquid mulch film. However, the preparation of the aforementioned mulch films still faces some technical and economic challenges, including improving the mechanical strength of biodegradable mulch films, controlling product breakage and degradation, enhancing their ability to increase temperature and retain moisture, and improving soil microbial diversity. Therefore, mulch films that are environmentally friendly and can improve soil microbial diversity urgently need further research. Summary of the Invention

[0004] The purpose of this disclosure is to provide a microbial preparation for improving soil and increasing productivity, and its liquid mulch application. This microbial preparation is easy to use, green and pollution-free, and can increase the biological yield of crops, increase the water content in the soil, improve the community structure of microorganisms in the soil, and promote the expansion of probiotic niches.

[0005] To achieve the above objectives, on the one hand, this disclosure provides a microbial preparation containing Rhizopus oligosporus ferment, Bacillus pumilus ferment, and carboxymethyl cellulose;

[0006] The Rhizopus arrhizus is classified as Rhizopus arrhizus, with accession number CGMCC No. 40610; the Bacillus pumilus is classified as Bacillus pumilus, with accession number CGMCC No. 13208.

[0007] Optionally, in the microbial preparation, the content of the Bacillus pumilus ferment is 10-50 parts by weight relative to 10-50 parts by weight of Rhizopus spp. ferment, and the content of the carboxymethyl cellulose is 0.1-10 parts by weight.

[0008] Preferably, the content of the Rhizopus oryzae ferment is 10-50 parts by weight relative to 100 parts by weight of the microbial preparation.

[0009] Optionally, the Rhizopus oligosporus ferment is obtained by inoculating Rhizopus oligosporus into a first culture medium and carrying out a first fermentation; preferably, the viable count in the Rhizopus oligosporus ferment is 150-1000 CFU / g, more preferably 300-600 CFU / g; the Bacillus pumilus ferment is obtained by inoculating Bacillus pumilus into a second culture medium and carrying out a second fermentation; preferably, the viable count in the Bacillus pumilus ferment is 150-1000 CFU / g, more preferably 300-600 CFU / g; the average particle size of the microbial preparation is 0.2-5.0 μm.

[0010] On the other hand, this disclosure provides a liquid mulch film for improving soil and increasing productivity, the liquid mulch film containing the aforementioned microbial agent and water; the amount of the microbial agent is 0.1 to 1 kg per 100 kg of water.

[0011] On the other hand, this disclosure provides the application of the aforementioned microbial preparations in increasing the biological yield of crops.

[0012] Optionally, the method includes the following steps: mixing the microbial agent with water and spraying it onto the soil surface after crop sowing; preferably, the amount of the microbial agent used is 0.1 to 1 kg relative to 100 kg of water; preferably, the crop includes at least one of wheat, corn, soybean and potato.

[0013] On the other hand, this disclosure provides the application of the aforementioned microbial agents in increasing soil moisture content and temperature.

[0014] Optionally, the method includes the following steps: mixing the microbial preparation with water and spraying it onto the soil surface; preferably, the amount of the microbial preparation used is 0.1 to 1 kg relative to 100 kg of water.

[0015] On the other hand, this disclosure provides the application of the aforementioned microbial agents in regulating the structure of soil microbial communities.

[0016] Optionally, the method includes the following steps: mixing the microbial preparation with water and spraying it onto the soil surface; preferably, the amount of the microbial preparation used is 0.1 to 1 kg relative to 100 kg of water; preferably, the soil microorganisms include at least one of the following: Acidobacteria, Green Curvularia, Bacillus, and Actinomycetes.

[0017] Through the above technical solution, this disclosure provides a microbial agent for improving soil and increasing productivity, and its liquid mulch application. On the one hand, the raw materials of the microbial agent are natural, the microbial agent is green and pollution-free, does not require recycling after use, and is simple to use. On the other hand, after the microbial agent is applied to the soil surface after crop sowing, the microbial agent can increase the water content in the soil, regulate the community structure of microorganisms in the soil, and promote the expansion of the ecological niche of probiotics.

[0018] Other features and advantages of this disclosure will be described in detail in the following detailed description section.

[0019] Information on the preservation of biological materials

[0020] The taxonomic name of Rhizopus arrhizus is Rhizopus arrhizus. It is deposited at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is April 26, 2023, and the accession number is CGMCC No. 40610.

[0021] The taxonomic name of the Bacillus pumilus is Bacillus pumilus. It is deposited at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is October 27, 2016, and the accession number is CGMCC No. 13208. Attached Figure Description

[0022] The accompanying drawings are provided to further illustrate the present disclosure and form part of the specification. They are used together with the following detailed description to explain the present disclosure, but do not constitute a limitation thereof. In the drawings:

[0023] Figure 1 This is a graph showing the above-ground fresh weight of wheat in the experimental and control groups.

[0024] Figure 2 This is a graph showing the dry weight of wheat grains in the experimental and control groups.

[0025] Figure 3 This is a graph showing the thousand-grain weight of wheat in the experimental and control groups.

[0026] Figure 4 Curves of accumulated temperature during the growing season for the soil in the experimental and control groups.

[0027] Figure 5 Curves showing the average soil moisture content during the growing season for the experimental and control groups. Detailed Implementation

[0028] The specific embodiments of this disclosure will be described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described herein are for illustration and explanation only and are not intended to limit this disclosure.

[0029] On the one hand, this disclosure provides a microbial preparation containing Rhizopus oligosporus ferment, Bacillus pumilus ferment, and carboxymethyl cellulose;

[0030] The Rhizopus arrhizus is classified as Rhizopus arrhizus and its accession number is CGMCC No. 40610. The Bacillus is classified as Bacillus pumilus and its accession number is CGMCC No. 13208.

[0031] In this disclosure, Rhizopus septemlobus ferment and Bacillus pumilus ferment are used. These two ferments have the advantages of high viable count, high density and few contaminants. When applied to soil, Rhizopus septemlobus ferment and Bacillus pumilus ferment can give full play to their characteristics of rapid reproduction and strong vitality.

[0032] In this disclosure, both Rhizopus oligosporus and Bacillus pumilus are deposited at the China General Microbiological Culture Collection Center.

[0033] In one specific embodiment of this disclosure, the microbial preparation contains 10-50 parts by weight of Bacillus pumilus ferment relative to 10-50 parts by weight of Rhizopus spp. ferment, and 0.1-10 parts by weight of carboxymethyl cellulose ferment.

[0034] In a preferred embodiment of this disclosure, the content of the Rhizopus oryzae ferment is 10-50 parts by weight relative to 100 parts by weight of the microbial preparation.

[0035] In one specific embodiment of this disclosure, the Rhizopus spp. fermentation product is obtained by inoculating Rhizopus spp. into a first culture medium and carrying out a first fermentation.

[0036] In the above embodiments, the first fermentation may include a first liquid fermentation and a first solid fermentation, preferably the first solid fermentation; specifically, the conditions for the first liquid fermentation include: a temperature of 25-30°C and a time of 2-6 days. The conditions for the first solid fermentation include: a temperature of 25-30°C and a time of 7-14 days. The first culture medium may include a first liquid culture medium and a first solid culture medium, wherein the first liquid culture medium is selected from one of LB, PDB, YS, and YPD; and the first solid culture medium is selected from one or two of PDA, rice husk powder, rice grains, corn flour, and biochar.

[0037] In one specific embodiment of this disclosure, the viable count in the Rhizopus oryzae ferment is 150–1000 CFU / g, preferably 300–600 CFU / g.

[0038] In one embodiment of this disclosure, the Bacillus pumilus ferment is obtained by inoculating Bacillus pumilus into a second culture medium and carrying out a second fermentation.

[0039] In the above embodiments, the second fermentation may include a second liquid fermentation and a second solid fermentation, preferably a second solid fermentation; specifically, the conditions for the second liquid fermentation include: a temperature of 25–30°C and a time of 2–6 days. The conditions for the second solid fermentation include: a temperature of 25–30°C and a time of 7–14 days. The second culture medium may include a second liquid culture medium and a second solid culture medium, wherein the second liquid culture medium is selected from one of LB, PDB, YS, and YPD; and the second solid culture medium is selected from one or two of PDA, rice husk powder, rice grains, corn flour, and biochar.

[0040] In one specific embodiment of this disclosure, the viable count in the Bacillus pumilus ferment is 150–1000 CFU / g, preferably 300–600 CFU / g.

[0041] In one specific embodiment of this disclosure, the average particle size of the microbial preparation is 0.2–5.0 μm.

[0042] In this disclosure, the microbial preparation contains Rhizopus oligosporus ferment, Bacillus pumilus ferment, and carboxymethyl cellulose. The preparation method of the microbial preparation includes mixing the Rhizopus oligosporus ferment, Bacillus pumilus ferment, and carboxymethyl cellulose, and then subjecting the mixed material to ultrafine pulverization to obtain a microbial preparation capable of improving soil microbial diversity and productivity, named "ZWLFmic". The average particle size of this microbial preparation can be 0.2–5.0 μm. The "ultrafine pulverization" can be performed using instruments or methods conventional in the art, and will not be elaborated further here.

[0043] On the other hand, this disclosure provides a liquid mulch film for improving soil and increasing productivity, the liquid mulch film containing the aforementioned microbial agents and water.

[0044] In one specific embodiment of this disclosure, the amount of the microbial preparation used is 0.1 to 1 kg per 100 kg of water.

[0045] On the other hand, this disclosure provides the application of the aforementioned microbial preparations in increasing the biological yield of crops.

[0046] In one specific embodiment of this disclosure, the method includes the following steps: mixing the microbial agent with water and spraying it onto the soil surface after crop sowing; in a preferred embodiment of this disclosure, the amount of the microbial agent used is 0.1 to 1 kg relative to 100 kg of water; in a preferred embodiment of this disclosure, the crop includes at least one of wheat, corn, soybean and potato.

[0047] On the other hand, this disclosure provides the application of the aforementioned microbial agents in increasing soil moisture content and temperature.

[0048] In one specific embodiment of this disclosure, the method includes the following steps: mixing the microbial agent with water and spraying it onto the soil surface; in a preferred embodiment of this disclosure, the amount of the microbial agent used is 0.1 to 1 g relative to 100 kg of water.

[0049] On the other hand, this disclosure provides the application of the aforementioned microbial agents in regulating the structure of soil microbial communities.

[0050] In one specific embodiment of this disclosure, the method includes the following steps: mixing the microbial preparation with water and spraying it onto the soil surface; in a preferred embodiment of this disclosure, the amount of the microbial preparation used relative to 100 kg of water is 0.1–1 g; in a preferred embodiment of this disclosure, the soil microorganisms include at least one of Acidobacteria bacterium, Chloroflexi bacterium, Gemmatimonadetes bacterium, and Actinomycetia bacterium.

[0051] In this disclosure, the "spraying" described in the above embodiments can be performed using instruments or methods conventional in the art, such as sprinkler irrigation technology or a handheld sprayer.

[0052] The present disclosure is further described in detail below through examples. All raw materials used in the examples are commercially available.

[0053] The model of the ultrafine pulverizer is QF70.

[0054] The model number of the handheld sprayer is 3WBD-20-4.

[0055] Example 1

[0056] This example illustrates the preparation of microbial preparations:

[0057] Preparation of Rhizopus septemlobus ferment: Rhizopus septemlobus was inoculated into YS medium and cultured at 25-30℃ for 4 days.

[0058] Preparation of Bacillus pumilus ferment: Bacillus pumilus was inoculated into LB medium and cultured at 25–30°C for 2 days.

[0059] 25 kg of Rhizopus spp. fermentation product and 25 kg of Bacillus pumilus fermentation product were mixed together and then mixed with 1 kg of carboxymethyl cellulose. The mixed material was then subjected to ultra-micro pulverization using an ultra-micro pulverizer to obtain a microbial preparation with an average particle size of 3 μm and a viable count of 600 CFU / g.

[0060] Example 2

[0061] This example illustrates the method of using microbial preparations:

[0062] The microbial preparations from Example 1 were weighed in batches of 0.5 kg (T1), 1.0 kg (T2), 2.0 kg (T3), and 3.0 kg (T4), respectively. They were then dissolved in 300 L of water. After the microbial preparations were completely dissolved, they were shaken well and sprayed onto the soil surface after wheat sowing using a handheld sprayer. T1, T2, T3, and T4 were the experimental groups, while the control group (CK) received no treatment.

[0063] Test Example 1

[0064] This test case illustrates the effect of microbial agents on wheat biological yield:

[0065] (1) Method for testing the fresh weight of wheat aboveground parts: After wheat is harvested, all harvested parts are weighed and the weight is calculated based on the unit area.

[0066] The calculation formula is: aboveground fresh weight (kg / hm) 2 = Harvested wheat weight (kg) / Area of ​​land used for wheat (hm²) 2 ).

[0067] (2) Method for testing the dry weight of wheat grains: After the wheat is dried and threshed, the weight of the wheat grains is weighed and the weight is calculated based on the unit area; the drying conditions include: temperature of 45-55℃ and time of 24-72h.

[0068] The calculation formula is: Dry weight of grains per ear (kg / hm) 2 = Weight of harvested wheat after drying and threshing (kg) / Area of ​​land used for wheat cultivation (hm²) 2 ).

[0069] (3) Method for testing the weight of 1,000 wheat grains: Randomly select 1,000 dried wheat grains and weigh them.

[0070] The results of wheat aboveground fresh weight, wheat ear dry weight, and wheat thousand-grain weight are as follows: Figures 1-3 As shown in Table 1, the productivity effect of wheat is characterized by the average growth rate of wheat aboveground fresh weight, wheat ear dry weight, and wheat thousand-grain weight.

[0071] Average growth rate (%) = (experimental group - control group) / blank control group × 100%.

[0072] Table 1

[0073] Fresh weight of aboveground parts 0 10 29 37 35 Dry weight of grains per ear 0 20 23 30 39 1000 grains weight 0 6 9 8 7

[0074] Depend on Figures 1-3 As can be seen from the data in Table 1, the microbial preparations prepared in this disclosure can increase the biological yield of wheat.

[0075] Test Example 2

[0076] This test case illustrates the effects of microbial agents on soil moisture content and soil temperature.

[0077] (1) Soil temperature test method: Soil temperature tester, use according to the specifications and instructions of different manufacturers.

[0078] (2) Soil moisture content test method: Soil moisture tester, use according to the specifications and instructions of different manufacturers.

[0079] During the wheat growing season, the accumulated temperature and average soil moisture content during the growing season are as follows: Figure 4 and 5 As shown in Table 2, the changes in soil water temperature are represented by the accumulated temperature during the growing season and the average growth rate of soil moisture content during the growing season.

[0080] Average growth rate (%) = (experimental group - control group) / blank control group × 100%.

[0081] Table 2

[0082] Accumulated temperature during the growing season 0 9.8 10.9 13.1 16.0 Average soil moisture content during the growing season 0 12.6 16.0 20.2 26.6

[0083] Depend on Figure 4 and 5 As can be seen from the data in Table 1, the microbial preparations prepared in this disclosure can increase the accumulated temperature and average soil moisture content during the wheat growing season.

[0084] Test Example 3

[0085] This test case illustrates the impact of microbial agents on soil microbial community structure:

[0086] (1) Testing methods for soil microbial species: 16s high-throughput sequencing combined with metagenomic high-throughput detection. Analysis of variance of soil microbial species is listed in Table 3.

[0087] (2) Methods for detecting bacterial species in soil: 16s high-throughput sequencing combined with metagenomic high-throughput detection. Analysis of variance of soil bacterial species is listed in Table 4.

[0088] (3) Test method for actinomycetes in soil: 16s high-throughput sequencing combined with metagenomic high-throughput detection. The variance analysis of soil actinomycetes is listed in Table 5.

[0089] Table 3

[0090]

[0091] Table 4

[0092]

[0093] Table 5

[0094]

[0095] Table 3 shows that the diversity of microbial species in the soil during the wheat growing season was significantly different (p = 0.02 < 0.05). Tables 4 and 5 show that the microbial agent mainly affects the composition and structure of soil microorganisms, primarily the actinomycete community.

[0096] The preferred embodiments of this disclosure have been described in detail above with reference to the accompanying drawings. However, this disclosure is not limited to the specific details of the above embodiments. Within the scope of the technical concept of this disclosure, various simple modifications can be made to the technical solutions of this disclosure, and these simple modifications all fall within the protection scope of this disclosure.

[0097] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, this disclosure will not describe the various possible combinations separately.

[0098] Furthermore, various different embodiments of this disclosure can be combined in any way, as long as they do not violate the spirit of this disclosure, they should also be regarded as the content disclosed in this disclosure.

Claims

1. A microbial preparation, characterized in that, This microbial preparation contains Rhizopus oligosporus ferment, Bacillus pumilus ferment, and carboxymethyl cellulose; The classification name of the Rhizopus oligospermum is Rhizopus oligospermum. Rhizopusarrhizus The accession number is CGMCC No. 40610; the classified name of the described Bacillus pumilus is Bacillus pumilus. Bacillus pumilus The accession number is CGMCCNo.13208; In the microbial preparation, relative to 10-50 parts by weight of Rhizopus spp. fermentation product, the content of Bacillus pumilus fermentation product is 10-50 parts by weight, and the content of carboxymethyl cellulose is 0.1-10 parts by weight. The viable count of the Rhizopus spp. fermentation product is 150-1000 CFU / g; the viable count of the Bacillus pumilus fermentation product is 150-1000 CFU / g. The average particle size of the microbial preparation is 0.2~5.0 μm.

2. The microbial preparation according to claim 1, wherein, The content of the Rhizopus oryzae ferment is 10-50 parts by weight relative to 100 parts by weight of the microbial preparation.

3. The microbial preparation according to claim 1, wherein, The Rhizopus septemlobus ferment is obtained by inoculating Rhizopus septemlobus into a first culture medium and carrying out a first fermentation. The viable count of the Rhizopus spp. fermentation product is 300-600 CFU / g; The Bacillus pumilus ferment is obtained by inoculating Bacillus pumilus into a second culture medium and carrying out a second fermentation. The viable count in the Bacillus pumilus ferment is 300-600 CFU / g.

4. A liquid mulch film for improving soil and increasing productivity, characterized in that, The liquid mulch film contains the microbial preparation as described in any one of claims 1 to 3 and water; The dosage of the microbial preparation is 0.1 to 1 kg per 100 kg of water.

5. The application of the microbial preparation according to any one of claims 1-3 in improving the biological yield of crops.

6. The application according to claim 5, wherein, Includes the following steps: The microbial preparation is mixed with water and sprayed onto the soil surface after crop sowing.

7. The application according to claim 6, wherein, The amount of the microbial preparation used relative to 100 kg of water is 0.1~1 kg; The crops include at least one of wheat, corn, soybeans, and potatoes.

8. The use of the microbial preparation according to any one of claims 1-3 in improving soil moisture content and temperature.

9. The application according to claim 8, wherein, Includes the following steps: The microbial preparation is mixed with water and then sprayed onto the soil surface.

10. The application according to claim 9, wherein, The amount of the microbial preparation used is 0.1 to 1 kg relative to 100 kg of water.

11. The application of the microbial preparation according to any one of claims 1-3 in regulating the structure of soil microbial communities.

12. The application according to claim 11, wherein, Includes the following steps: The microbial preparation is mixed with water and then sprayed onto the soil surface.

13. The application according to claim 12, wherein, The amount of the microbial preparation used relative to 100 kg of water is 0.1~1 kg; The soil microorganisms include at least one of the following: Acidobacteria, Green Curvularia, Bacillus, and Actinomycetes.

Images