Microbial composition and application thereof in promoting rice growth

Abstract

The invention discloses a microbial composition. The microbial composition is glomus intraradices and burkholderia. The glomus intraradices is Glomus intraradices AMF-12, the classification name of the glomus intraradices AMF-12 is Glomus intraradices, the glomus intraradices AMF-12 is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number of the glomus intraradices AMF-12 is CGMCC No.41715, and the preservation date of the glomus intraradices AMF-12 is December 11, 2024; the Burkholderia is Burkholderia sp.Mir-06, the classification name of the Burkholderia sp.Mir-06 is Burkholderia sp.Mir-06, the Burkholderia is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number of the Burkholderia is CGMCC No.33223, and the preservation date of the Burkholderia is December 27, 2024; the invention further discloses application of the microbial composition in promoting rice growth. According to the method, glomus intraradices and burkholderia are jointly applied to treatment of rice seeds or applied to rice field soil for planting rice as microbial fertilizers, and the plant height, dry matter weight and yield of rice can be effectively improved, so that rice growth is promoted.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a microbial composition and application thereof in promoting rice growth. Background Art

[0002] As an environmentally friendly technology, microbial technology can gradually reduce the use of chemical products without affecting the yield and quality of crops, and has shown great application potential in the sustainable development of agriculture. In the late 19th century, researchers in the field of microbiology, led by Pasteur of France, achieved a series of results, which promoted the development and application of microbial research in the agricultural field, and thus produced microbial fertilizers with nitrogen fixation, phosphorus dissolution, potassium dissolution and other effects. At present, more than 70 countries in the world are researching, producing and using microbial fertilizers, such as the United States, France, India, and some African countries. Their varieties are mainly rhizobium preparations and bioremediation preparations, especially rhizobium preparations, which have developed most rapidly. Not only has the inoculation area been continuously expanding, but also a wide variety of legumes have been applied. Many countries have also made achievements in the research and application of other types of microorganisms. For example, researchers in the Soviet Union and some Eastern European countries have found that these bacteria can secrete growth substances and an antifungal antibiotic to promote plant seed germination and root growth. According to surveys, the application of microbial fertilizers in developed countries has accounted for more than 40% of the total fertilizer application, and shows an upward trend of 10% to 20% each year.

[0003] Arbuscular mycorrhizal fungi (AMF) are the most widely distributed fungi in the soil, which can form a symbiotic body with the roots of plants - arbuscular mycorrhiza. Arbuscular mycorrhiza can not only help plants absorb water and mineral salts in the soil, improve plant resistance to drought and heat, and nutritional conditions, but also improve soil characteristics, and play an important role in ecological environmental protection. Especially today, when the contradiction between man and land is becoming increasingly severe, AMF will inevitably have a broader development space and application prospects in agricultural production and ecological environmental restoration. At the same time, mycorrhizal symbiosis is conducive to the enrichment of Burkholderia, which is a kind of associative nitrogen-fixing bacteria with strong nitrogen-fixing ability. It has the potential to promote plant nitrogen utilization efficiency and plant growth as a biological fertilizer.

[0004] At present, the joint application of Burkholderia and AMF is mainly focused on vegetables. Burkholderia and AMF can synergistically promote the growth of vegetables. However, their joint application on rice has not yet been disclosed. Therefore, it is of great significance to discover the root spherical mold with higher growth promotion potential and Burkholderia with higher nitrogen fixation ability, and to mix them to form a microbial agent that can promote rice growth based on the interaction between the two in the ecological field. Summary of the invention

[0005] In view of the above problems, the present invention aims to provide a microbial composition and its application in promoting rice growth.

[0006] According to a first aspect of the present invention, there is provided a microbial composition comprising Glomus intraradices and Burkholderia;

[0007] The intraradical Glomus is Glomus intraradices AMF-12, classified and named Glomus intraradices, deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with a deposit number of CGMCC No.41715 and a deposit date of December 11, 2024;

[0008] The Burkholderia is Burkholderia sp. Mir-06, classified and named Burkholderia sp., deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC No. 33223 and the deposit date December 27, 2024.

[0009] According to a second aspect of the present invention, there is provided a product comprising the microbial composition or a fermentation product thereof according to the first aspect, wherein the product is any one of a seed soaking solution, a seed coating or a microbial fertilizer.

[0010] In some embodiments, the total viable count of the microbial composition in the product is not less than 10 8 CFU / g or 10 8 CFU / mL, the ratio of the viable counts of Glomus root fungus and Burkholderia is 1 to 3:1 to 3. For example, the ratio of the viable counts of Glomus root fungus and Burkholderia is 1:1, 1:2, 1:3, 3:1, and 2:1.

[0011] According to the third aspect of the present invention, there is provided a use of the microbial composition or the product in promoting rice growth.

[0012] In some implementations, the above application may include one or more of the following application combinations:

[0013] (1) Improve the application of high quality rice plants;

[0014] (2) Application in increasing the dry matter weight of rice;

[0015] (3) Application in increasing rice yield.

[0016] In some embodiments, the total viable count of the microbial composition is not less than 10 8 CFU / g or 108 CFU / mL, the ratio of the viable counts of Glomus root fungus and Burkholderia is 1-3:1-3, for example, the ratio of the viable counts of Glomus root fungus and Burkholderia is 1:1, 1:2, 1:3, 3:1, and 2:1.

[0017] According to a fourth aspect of the present invention, there is provided a use of the microbial composition or the product in improving the low nitrogen tolerance of rice.

[0018] According to a fifth aspect of the present invention, there is provided a method for culturing the microbial composition, comprising the following steps: applying intraradical spherical mold to sandy soil where tomatoes or clover are grown and culturing to the late reproductive stage, eluting spores and hyphae in the sand with a PBS reagent, and centrifuging to obtain bacterial mud; inoculating Burkholderia in LB culture medium, culturing at 28°C for 48 hours, collecting bacterial cells and extracellular polysaccharides, and centrifuging to obtain bacterial mud; mixing the two bacterial muds, diluting with physiological saline (0.9%), and preparing a seed soaking solution.

[0019] In some embodiments, the formula of the LB medium is: 10 g of tryptone, 5 g of yeast extract, 10 g of sodium chloride, 15 g of fat powder, add ddH2O to 1000 mL, and adjust the pH to 7.2 with 5 mol / L NaOH (about 0.2 mL).

[0020] According to a sixth aspect of the present invention, there is provided a method for processing rice seeds, comprising the following steps:

[0021] The rice seeds are soaked in a soaking solution obtained by fermenting and diluting the microbial composition described in the first aspect, and then the soaked rice seeds are coated with a coating agent prepared by mixing activated carbon and Trichoderma harzianum powder.

[0022] In some embodiments, the treatment method comprises the following steps:

[0023] S1. Preparation of seed soaking solution: Glomus intraradices AMF-12 was added to the sandy soil where tomatoes or clover were planted and cultured until the late reproductive stage, spores and hyphae in the sand were washed out with PBS reagent, and bacterial mud was obtained by centrifugation; Burkholderia sp. Mir-06 was inoculated into LB medium, cultured at 28°C for 48 hours, bacterial cells and extracellular polysaccharides were collected, and bacterial mud was obtained by centrifugation; the two bacterial muds were mixed and diluted with 0.9% physiological saline to prepare seed soaking solution;

[0024] Among them, the total number of live bacteria after mixing the two kinds of mud is not less than 10 9 CFU / g, the ratio of the number of viable bacteria of Glomus intraradices and Burkholderia spp. was 1-3:1-3;

[0025] S2. Soaking seeds: soaking rice seeds with the soaking solution, removing and draining to obtain soaked rice seeds;

[0026] Among them, the soaking time is 10 to 30 minutes;

[0027] S3. Preparation of coating agent: Mixing activated carbon and Trichoderma harzianum powder to prepare a coating agent;

[0028] The weight ratio of activated carbon to Trichoderma harzianum powder is 10 to 15:1, for example, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, etc. The total number of viable bacteria in the coating agent is not less than 10 9 CFU / g.

[0029] S4. Coating coating agent: adding the coating agent to the soaked rice seeds, mixing, and drying;

[0030] The weight ratio of the soaked rice seeds to the coating agent is 100-120:1, for example, 100:1, 105:1, 110:1, 115:1, 120:1; and the thickness of the coating agent on the surface of the seeds is 0.3-0.6 mm.

[0031] According to a seventh aspect of the present invention, there is provided a method for preparing and using a microbial fertilizer, comprising the following steps:

[0032] The microbial composition described in the first aspect or mixed with other microbial preparations to prepare a microbial fertilizer is applied to the paddy soil in the early stage of rice growth. (The microbial composition is mixed with sandy soil, and then the sandy soil is applied to the paddy soil where rice is planted)

[0033] In some embodiments, the method for using the microbial fertilizer comprises the following steps:

[0034] S1. Preparation of microbial fertilizer: Glomus intraradices AMF-12 is added to the sandy soil where tomatoes or clover are grown and cultured until the late reproductive stage, spores and hyphae in the sand are washed out with PBS reagent, and the bacterial sludge is obtained by centrifugation; Burkholderia sp. Mir-06 is inoculated into LB medium, cultured at 28°C for 48 hours, bacterial cells and extracellular polysaccharides are collected, and the bacterial sludge is obtained by centrifugation; the two bacterial sludges are mixed and prepared into microbial fertilizer directly or mixed with other microbial preparations;

[0035] Among them, the total number of live bacteria after mixing the two kinds of mud is not less than 10 9 CFU / g, the ratio of the number of viable bacteria of Glomus intraradices and Burkholderia spp. was 1-3:1-3;

[0036] S2. Application of microbial fertilizer: When planting rice, apply it to the paddy field soil by mixing with sand during tillage, seedling stage or transplanting. The earlier the application, the more conducive it is to the colonization, enrichment and growth promotion of Glomus intraradices and Burkholderia in the rice root system;

[0037] Among them, the weight ratio of sand and fungus mud is 1000-1500:1, the weight of the mixture of sand and fungus mud per mu of land is 2-3 kilograms, and the time of application is before planting, seedling stage or transplanting stage.

[0038] It should be noted that low nitrogen tolerance, plant height, dry matter weight and yield are all terms known in the art and have no other special meanings.

[0039] The beneficial effects of the present invention are:

[0040] 1. The present invention applies Glomus intraradicis and Burkholderia together in the treatment of rice seeds or applies them as microbial fertilizers to the soil of rice fields where rice is planted, which can effectively increase the plant height, dry matter weight and yield of rice, thereby promoting rice growth.

[0041] 2. The present invention applies Glomus intraradicis and Burkholderia together in the treatment of rice seeds or applies them as microbial fertilizers to the soil of rice fields where rice is planted, which can effectively improve the low nitrogen tolerance of rice, thereby reducing the use of nitrogen fertilizers, which is economical and environmentally friendly. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 The composition of rhizosphere soil microorganisms of rice with different mycorrhizal symbiosis efficiencies;

[0043] Figure 2 The rice yield after application of different AMF strains;

[0044] Figure 3 The mycorrhizal symbiosis of rice roots treated with different AMF strains;

[0045] Figure 4 It is the phylogenetic map of nitrogen-fixing bacteria isolated from the rhizosphere;

[0046] Figure 5 is the nitrogenase activity of the isolated nitrogen-fixing bacteria;

[0047] Figure 6It is a comparison chart of rice growth under different seed treatment conditions, wherein A is the rice growth of the mixed seed soaking and coating group in Example 1, B is the rice growth of the microbial fertilizer application group in Example 2, C is the rice growth of the blank control group in Comparative Example 1, D is the rice growth of the mixed seed soaking group in Comparative Example 2, E is the rice growth of the coating group in Comparative Example 3, F is the rice growth of the root spherical mold AMF-12 soaking and coating group in Comparative Example 4, G is the rice growth of the Burkholderia Mir-06 soaking and coating group in Comparative Example 5, and H is the rice growth of the conventional fertilization treatment group in Comparative Example 6. DETAILED DESCRIPTION

[0048] The present invention will be further described in detail below in conjunction with the embodiments.

[0049] In the following examples, Glomus intraradices AMF-12 was collected by itself in Nanchang County, Nanchang City, Jiangxi Province, and was deposited in the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Administration, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, deposit number: CGMCC No.41715, deposit date: December 11, 2024, classification name: Glomus intraradices.

[0050] The Burkholderia is Burkholderia sp. Mir-06, which was collected by itself in Nanchang County, Nanchang City, Jiangxi Province. It is deposited in the General Microbiology Center (CGMCC) of the China Microorganism Culture Collection Administration, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, deposit number: CGMCC No.33223, deposit date: December 27, 2024, classification name: Burkholderia sp.

[0051] In the following embodiments, "aerial part" refers to the part of rice growing out of the soil; "heading stage" refers to the period when 80% of rice plants have heading; "maturity stage" refers to the period when 95% of the husks of rice ears turn yellow; "plant height" refers to the distance from the base of the stem to the top of the ear; "dry matter weight" refers to the constant weight of the above-ground part of rice after drying at 80°C for a certain period of time; and "yield" refers to the weight of grains with 13% moisture content of a single rice plant.

[0052] 1. Burkholderia cerevisiae and AMF synergistically promote growth

[0053] By measuring the composition of rhizosphere soil microorganisms in rice with different mycorrhizal symbiosis efficiencies, it was found that the rhizosphere microbial communities of rice with more efficient mycorrhizal symbiosis (A7 in Nanchang base and C7 in Hainan base) contained a higher abundance of Burkholderia sp. ( Figure 1 ).

[0054] The results showed that the interaction between arbuscular mycorrhiza and rice can promote the enrichment of Burkholderia in the rhizosphere of rice. Burkholderia has strong nitrogen fixation activity. The two synergistically promote the absorption of nitrogen nutrients by rice, thereby promoting the growth and yield of rice.

[0055] 2. Isolation and screening of Glomus radicifolius AMF-12 and Burkholderia solani Mir-06

[0056] (1) Isolation and screening of root fungus AMF-12

[0057] In the rice low-nitrogen experimental field in Nanchang, Jiangxi, rhizosphere soil samples were collected at the heading stage of rice. The soil sample DNA was quickly extracted using a soil genomic DNA extraction kit, and then nested PCR amplification was performed using two pairs of primers AML1-AML2 and NS31-AM1 for arbuscular mycorrhizal fungi, and the amplified products were detected by agarose gel electrophoresis and fluorescence quantitative system. The PCR amplification products were sequenced by second-generation high-throughput sequencing to obtain arbuscular mycorrhizal fungi of different families and genera in the soil. Twenty dominant species of indigenous arbuscular mycorrhizal fungi were obtained using the biocommunity-related index. The above soil The spores of dominant species of arbuscular mycorrhizal fungi were collected, and the spores were further placed in a proper amount of river sand in a pot experiment, and various dominant species were propagated with the help of tomatoes and clover as hosts; the above-produced dominant species were inoculated into sterilized soil, and three-leaf rice seedlings were transplanted and cultured to maturity under the same culture conditions, and the rice yield was tested at maturity; according to the rice yield results tested above, the strains with higher rice yield were identified as efficient mycorrhizal symbiotic strains, which can promote the growth and yield of rice; according to the yield results, three strains, AMF-07, AMF-12 and AMF-18, were preliminarily screened out ( Figure 2 ); The symbiosis between rice roots and arbuscular mycorrhizal fungi at the heading stage was tested for the three selected fungi, with the detection indicators including mycorrhizal colonization frequency (F%) and mycorrhizal intensity (M%); the test results showed that the mycorrhizal colonization frequency and mycorrhizal intensity of AMF-12 in rice roots were both high ( Figure 3 ). Based on the above experimental results, it is clear that the root spherical fungus AMF-12 is the most efficient arbuscular mycorrhizal fungus.

[0058] (2) Isolation and screening of Burkholderia Mir-06

[0059] In the rice low-nitrogen experimental field in Nanchang, Jiangxi, rhizosphere soil samples were collected at the heading stage of rice. PBS buffer was added to the soil samples for dilution, and the supernatant was spread on the nitrogen-free Ashby medium. The samples were placed in an incubator with their front side incubated at 28°C for 3-4 days, and turbid and translucent colonies appeared on the medium. A few of the above colonies were picked up with an inoculation loop, and separated by streaking on a nitrogen-free Ashby medium plate, and then incubated at 28°C for 3-4 days. A single colony appeared on the plate, which was aseptically transferred into a nitrogen-free Ashby medium slant test tube and incubated at 28°C for 4 days. By this step, a total of 100 nitrogen-fixing strains were isolated, and 16S rDNA detection revealed that all the isolated strains were Burkholderia, and through phylogenetic comparison, 13 distantly related Burkholderia strains R6, R7, R12, R13, R16, R17, R35, R40, R56, R62, R81, R104, R121, etc. were screened out ( Figure 4 ); add 4 mL of Ashby nitrogen fixation medium to a screw tube with a volume of 21 mL to make a slant, inoculate and culture in a 28°C incubator, use the uninoculated empty slant as a negative control, and the round brown ammonia-fixing bacteria with known enzyme activity as a positive control; after 72 hours, replace the rubber stopper, inject 10% ethane gas (prepared by calcium carbide), seal with a sealing film, and continue to culture for 72 hours; take 100 μL of the reaction gas to measure the ethylene production in a gas chromatograph, and convert it into ammonia fixation activity according to the following formula; screen strains with stronger nitrogenase activity according to the nitrogenase activity, and finally find that the nitrogenase activity of R6 is significantly higher than that of other strains, and it is named Mir-06.

[0060] Both Glomus root fungus AMF-12 and Burkholderia spp. Mir-06 were strains isolated from low-nitrogen rice fields. The isolated Glomus root fungus AMF-12 and Burkholderia spp. Mir-06 are more adaptable to the rice field environment and have a more obvious effect on promoting rice growth.

[0061] 3. Experimental Procedure

[0062] Step 1. Preparation of planting soil: Take paddy field soil, dry it in the sun, grind it into fine powder, and put it into the planting container as planting soil.

[0063] Step 2. Seed preparation: 400 rice seeds of uniform size and full grains were selected and randomly divided into 8 groups, each with 50 seeds. The 8 groups were mixed seed soaking and coating group, microbial fertilizer application group, blank control group, mixed seed soaking group, coating group, intraradical Glomus AMF-12 seed soaking and coating group, Burkholderia Mir-06 seed soaking and coating group and conventional fertilization treatment group.

[0064] Step 3. Seed coating:

[0065] Preparation of mixed soaking solution: Glomus intraradices (AMF-12) was added to the sandy soil where tomatoes or clover were planted and cultured until the late reproductive stage, spores and hyphae in the sand were washed with PBS reagent, and bacterial mud was obtained by centrifugation; Burkholderia sp. (Mir-06) was inoculated into LB medium and cultured at 28°C for 48 hours, bacterial cells and extracellular polysaccharides were collected, and bacterial mud was obtained by centrifugation; the bacterial mud of Glomus intraradices and Burkholderia sp. was diluted with sterile physiological saline (0.9%) to prepare Glomus intraradices and Burkholderia soaking solutions, respectively, and the total viable counts of the two species in the soaking solution were determined, and the soaking solutions of the two species were mixed to control the total viable count of each bacterium in the mixed soaking solution to be not less than 10 8 CFU / mL, the total number of viable bacteria after mixing with the soaking solution is 10 9 CFU / mL, the ratio of the live counts of the two bacteria is 1~2:1~2.

[0066] Preparation of coating agent: The activated carbon and Trichoderma harzianum powder were mixed in a weight ratio of 12:1 to prepare a coating agent, wherein the total number of viable bacteria was 10 9 CFU / g.

[0067] Seed coating: soak rice seeds in the prepared mixed soaking solution for 20 minutes, remove and drain; then add coating agent to the soaked rice seeds, the weight ratio of the soaked rice seeds to the coating agent is 100:1, the thickness of the coating agent on the seed surface is 0.4-0.6mm, mix well, and dry.

[0068] Step 4. Preparation and application of microbial fertilizer:

[0069] Preparation of microbial fertilizer: In the sandy soil where tomatoes or clover are planted, Glomus aestivum AMF-12 is applied and cultured until the late reproductive stage, spores and hyphae in the sand are washed off with PBS reagent, and bacterial mud is obtained by centrifugation; Burkholderia Mir-06 is inoculated into LB medium, cultured at 28°C for 48 hours, bacterial cells and extracellular polysaccharides are collected, and bacterial mud is obtained by centrifugation; Glomus aestivum and Burkholderia bacterial mud are diluted with sterile physiological saline (0.9%) to prepare Glomus aestivum and Burkholderia suspensions, respectively, and the total viable counts of the two strains in the soaking solution are determined, and the two strain suspensions are mixed, and the total viable counts of each strain in the mixed suspension are controlled to be not less than 10 8 CFU / mL, the total viable count after suspension mixing is 10 9 CFU / mL, the ratio of the live counts of the two bacteria is 1~2:1~2.

[0070] Application of microbial fertilizer: Add bacterial suspension to sandy soil at a weight ratio of 1000-1500:1 and stir evenly. Sprinkle the mixture of sandy soil and bacteria in the planting soil before transplanting, 0.05 kg per pot.

[0071] Step 5. Rice cultivation and growth data determination

[0072] 8 groups of rice seeds were treated respectively according to the methods of the following Examples 1-2 and Comparative Examples 1-6, and the 8 groups of rice seeds after treatment were planted in the same seedling substrates of different containers, cultured to the 3-leaf stage under the same light, temperature and humidity conditions, and then transplanted into the planting container, with 5 pots planted in each group. Among them, the microbial fertilizer application group was mixed with the prepared microbial fertilizer in the planting soil before transplanting. Rice was cultured in the planting container to maturity under the same light, temperature and humidity conditions. Except for the conventional fertilization treatment group of Comparative Example 6, which was 100% nitrogen application, the nitrogen application amount of the other groups was 75% of the nitrogen application amount of the conventional fertilization treatment group of Comparative Example 6, and the application amount of other fertilizers was consistent (specific fertilization conditions are shown in Table 1). The plant height of rice was measured at the rice heading stage, and the dry matter weight and yield of rice were measured at maturity.

[0073] Example 1

[0074] Mixing the soaking and coating group. Soak the rice seeds in the mixed soaking solution prepared in step 3 for 20 minutes, remove and drain; then add the coating agent to the soaked rice seeds, and the weight ratio of the soaked rice seeds to the coating agent is 100:1. The thickness of the coating agent on the surface of the seeds is 0.4-0.6 mm, mix well, and dry.

[0075] Example 2

[0076] Microbial fertilizer application group: Soak rice seeds in sterile water for 20 minutes, remove and drain; spread the microbial fertilizer prepared in step 4 in the planting soil before transplanting, 0.05 kg per pot.

[0077] Comparative Example 1

[0078] Blank control group: Soak rice seeds in sterile water for 20 minutes, then remove and drain.

[0079] Comparative Example 2

[0080] Mixed seed soaking group: Soak rice seeds in the mixed seed soaking solution prepared in step 3 for 20 minutes, remove and drain.

[0081] Comparative Example 3

[0082] Coating group: Soak rice seeds in sterile water for 20 minutes, remove and drain; then add coating agent to the soaked rice seeds, the weight ratio of soaked rice seeds to coating agent is 100:1. The thickness of the coating agent on the surface of the seeds is 0.4-0.6mm, mix well, and dry.

[0083] Comparative Example 4

[0084] Glomus intraradices AMF-12 seed soaking coating group. Soak rice seeds in Glomus intraradices AMF-12 soaking solution for 20 minutes, remove and drain; then add coating agent to the soaked rice seeds, and the weight ratio of rice seeds to coating agent after soaking is 100:1. The thickness of the coating agent on the surface of the seeds is 0.4-0.6mm, mix well, and dry.

[0085] Comparative Example 5

[0086] Burkholderia Mir-06 seed soaking coating group. Soak rice seeds in Burkholderia Mir-06 soaking solution for 20 minutes, remove and drain; then add coating agent to the soaked rice seeds, and the weight ratio of rice seeds to coating agent after soaking is 100:1. The thickness of the coating agent on the surface of the seeds is 0.4-0.6mm, mix well, and dry.

[0087] Comparative Example 6

[0088] Conventional fertilization treatment group. Seed treatment was the same as that of Comparative Example 1.

[0089] Table 1 Fertilizer application amount (mg / pot) of each embodiment and comparative example

[0090]

[0091] Test results: The relevant test results are shown in Table 2, and the rice growth conditions of each group are compared ( Figure 6 ).

[0092] Table 2 Plant height, dry matter weight and yield of rice in different treatments

[0093]

[0094] Note: Different lowercase letters indicate significant differences within the column (P<0.05).

[0095] It can be seen from the experimental results that: compared with comparative example 1, the mixed seed soaking and coating group of Example 1 and the microbial fertilizer application group of Example 2 can significantly increase the plant height, dry matter weight and yield of rice, and the microbial fertilizer application group of Example 2 is slightly better than the mixed seed soaking and coating group of Example 1; the phenotypic indicators of Example 1 are higher than those of comparative examples 2-5, and there are significant differences between the phenotypic indicators of Example 1 and comparative examples 3 and 5; there are no significant differences between the phenotypic indicators of Example 2 and comparative example 6. In general, the various indicators of the mixed seed soaking and coating group of Example 1 are better than those of comparative examples 2-5. When the nitrogen application rate is reduced by 25%, the plant height, dry matter weight and yield of the microbial fertilizer application group of Example 2 can reach the level of conventional nitrogen application treatment in comparative example 6.

[0096] The above are only some embodiments of the present invention. For those skilled in the art, several modifications and improvements may be made without departing from the inventive concept of the present invention, which all fall within the protection scope of the present invention.

Claims

1. A microbial composition, characterized in that The microbial composition is Glomus intraradici and Burkholderia; The intraradical Glomus is Glomus intraradices AMF-12, classified and named Glomus intraradices, deposited in the General Microbiological Center of China Microbiological Culture Collection Administration, with a deposit number of CGMCC No.41715 and a deposit date of December 11, 2024; The Burkholderia is Burkholderia sp. Mir-06, classified and named Burkholderia sp., deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC No.33223 and the deposit date December 27, 2024.

2. A product comprising the microbial composition or a fermentation product thereof according to claim 1, wherein the product is any one of a seed soaking solution, a seed coating or a microbial fertilizer.

3. The product according to claim 2, characterized in that The total viable count of the microbial composition in the product is not less than 10 8 CFU / g or 10 8 CFU / mL, the ratio of the live counts of Aspergillus niger and Burkholderia spp. was 1-3:1-3.

4. Use of the microbial composition according to claim 1 or the product according to claim 2 in promoting rice growth.

5. The use according to claim 4, characterized in that The application is one or more of the following: (1) Improve the application of high quality rice plants; (2) Application in increasing the dry matter weight of rice; (3) Application in increasing rice yield.

6. The use according to claim 4, characterized in that The total viable count of the microbial composition is not less than 10 8 CFU / g or 10 8 CFU / mL, the ratio of the live counts of Aspergillus niger and Burkholderia spp. was 1-3:1-3.

7. A method for treating rice seeds, characterized in that: The processing method comprises the following steps: The rice seeds are soaked in a soaking solution obtained by fermenting and diluting the microbial composition according to claim 1, and then the soaked rice seeds are coated with a coating agent prepared by mixing activated carbon and Trichoderma harzianum powder.

8. The processing method according to claim 7, characterized in that: The processing method comprises the following steps: S1. Preparation of seed soaking solution: Glomus intraradices AMF-12 was added to the sandy soil where tomatoes or clover were planted and cultured until the late reproductive stage, spores and hyphae in the sand were washed out with PBS reagent, and bacterial mud was obtained by centrifugation; Burkholderia sp. Mir-06 was inoculated into LB medium, cultured at 28°C for 48 hours, bacterial cells and extracellular polysaccharides were collected, and bacterial mud was obtained by centrifugation; the two bacterial muds were mixed and diluted with 0.9% physiological saline to prepare seed soaking solution; Among them, the total number of live bacteria after mixing the two kinds of mud is not less than 10 9 CFU / g, the ratio of the number of viable bacteria of Glomus intraradices and Burkholderia spp. was 1-3:1-3; S2. Soaking seeds: soaking rice seeds with the soaking solution, removing and draining to obtain soaked rice seeds; Among them, the soaking time is 10 to 30 minutes; S3. Preparation of coating agent: Mixing activated carbon and Trichoderma harzianum powder to prepare a coating agent; The weight ratio of activated carbon to Trichoderma harzianum powder is 10-15:1, and the total number of live bacteria in the coating agent is not less than 10 9 CFU / g; S4. Coating coating agent: adding the coating agent to the soaked rice seeds, mixing, and drying; The weight ratio of the soaked rice seeds to the coating agent is 100-120:1, and the thickness of the coating agent on the surface of the seeds is 0.3-0.6 mm.

9. A method for applying microbial fertilizer, characterized in that: The application method comprises the following steps: The microbial composition according to claim 1 or a mixture thereof with other microbial preparations is used to prepare a microbial fertilizer, which is applied to paddy field soil in the early stage of rice growth.

10. The administration method according to claim 9, characterized in that The application method comprises the following steps: S1. Preparation of microbial fertilizer: Glomus intraradices AMF-12 is added to the sandy soil where tomatoes or clover are grown and cultured until the late reproductive stage, spores and hyphae in the sand are washed out with PBS reagent, and the bacterial sludge is obtained by centrifugation; Burkholderia sp. Mir-06 is inoculated into LB medium, cultured at 28°C for 48 hours, bacterial cells and extracellular polysaccharides are collected, and the bacterial sludge is obtained by centrifugation; the two bacterial sludges are mixed and prepared into microbial fertilizer directly or mixed with other microbial preparations; Among them, the total number of live bacteria after mixing the two kinds of mud is not less than 10 9 CFU / g, the ratio of the number of viable bacteria of Glomus intraradices and Burkholderia spp. was 1-3:1-3; S2. Application of microbial fertilizer: When planting rice, apply it to the paddy field soil by mixing with sand during tillage, seedling stage or transplanting. The earlier the application, the more conducive it is to the colonization, enrichment and growth promotion of Glomus intraradices and Burkholderia in the rice root system; Among them, the weight ratio of sand and bacterial mud is 1000-1500:1, and the weight of the mixture of sand and bacterial mud per acre is 2-3 kilograms.

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