A growth-promoting bacteria, composite bacteria, bacterial agent, bacterial fertilizer and application

Abstract

The application provides a kind of promoting bacteria, composite bacteria, bacterial agent, bacterial fertilizer and application, including pyrrocinia burkholderia and Pseudomonas atagonensis;The pyrrocinia burkholderia is Burkholderia pyrrocinia FJS-3, which has been preserved in Guangdong Microbial Culture Collection Center, with a preservation date of July 20, 2022, and a preservation number of GDMCC No.62432;The Pseudomonas atagonensis is Pseudomonas atagonensis FJS-16, which has been preserved in Guangdong Microbial Culture Collection Center, with a preservation date of July 20, 2022, and a preservation number of GDMCC No.62433.The promoting composite bacteria provided by the application has a better growth-promoting effect on plants.

Description

Technical Field

[0001] This invention relates to microorganisms and their applications, and more particularly to a growth-promoting bacterium, a compound bacterium, a microbial agent, a microbial fertilizer, and their applications. Background Technology

[0002] As a leaf crop, nitrogen nutrition is crucial for tea trees. In order to ensure tea production, producers tend to apply excessive nitrogen fertilizer. Over time, this leads to an imbalance in soil nutrient ratios, soil compaction, acidification, poor aeration, decreased root activity of tea trees, reduced number and activity of soil microorganisms, and reduced water and fertilizer retention capacity. Consequently, it seriously affects tea yield and quality, and weakens the tea tree's ability to resist diseases, pests, and adverse conditions.

[0003] Furthermore, due to declining soil quality, nitrogen and phosphorus are more easily lost, leading to a vicious cycle of continuously increasing fertilizer application and further exacerbating nutrient loss. This vicious cycle directly accelerates tea garden aging, which is extremely detrimental to its sustainable development. It also causes environmental pollution and eutrophication of water bodies. In the mountainous conditions of Guizhou, this loss of water, soil, and fertilizer will be even more severe. Therefore, the research and application of new technologies for high-quality tea garden soil and fertilizer management are urgently needed.

[0004] Guizhou is located on the eastern slope of the Yunnan-Guizhou Plateau, with widespread acidic yellow soil. The average altitude is around 1100 meters, the average annual temperature is between 10-20℃, and rainfall is abundant. Although Guizhou's tea gardens have advantages, improper production practices have led to problems such as soil nutrient imbalance, over-acidification, reduced biodiversity, and increased pests, diseases, and weeds. A survey of fertilization and soil fertility in Meitan County's tea gardens revealed that 84.4% of the fertilizers used were general-purpose fertilizers, with a single variety; nutrient ratios were unbalanced, with excessive nitrogen fertilizer application and a severe phosphorus deficiency in the tea garden soil.

[0005] In view of this, it is necessary to provide a growth-promoting bacterium, compound bacterium, bacterial agent, bacterial fertilizer and its application to solve or at least alleviate the technical defects of the above-mentioned plant growth being limited by traditional fertilizers. Summary of the Invention

[0006] The main objective of this invention is to provide a growth-promoting bacterium, a compound bacterium, a microbial agent, a microbial fertilizer, and their applications, aiming to solve the technical problem that plant growth is limited by traditional fertilizers in the prior art.

[0007] To achieve the above objectives, the present invention provides a growth-promoting bacterium, wherein the growth-promoting bacterium is Burkholderia pyrrole or Pseudomonas aeruginosa;

[0008] The Burkholderia pyrrocinia mentioned is Burkholderia pyrrocinia FJS-3, with the Latin taxonomic name Burkholderia pyrrocinia. It has been deposited at the Guangdong Provincial Center for Microbial Culture Collection on July 20, 2022, with the accession number GDMCC No. 62432 and the deposit address at No. 100, Xianlie Middle Road, Guangzhou.

[0009] The *Pseudomonas atagonensis* mentioned is *Pseudomonas atagonensis* FJS-16, with the Latin taxonomic name *Pseudomonas atagonensis*. It has been deposited at the Guangdong Provincial Center for Microbial Culture Collection on July 20, 2022, with the accession number GDMCC No. 62433, and the deposit address is No. 100, Xianlie Middle Road, Guangzhou.

[0010] The present invention also provides a growth-promoting compound bacteria, which includes Burkholderia pyrrole as described in any one of the above claims and Pseudomonas aeruginosa as described in any one of the above claims.

[0011] The present invention also provides a compound microbial agent, comprising the compound microorganisms as described in any one of the above claims.

[0012] Furthermore, the preparation process of the compound microbial agent includes:

[0013] Burkholderia pyrrocinia FJS-3 was inoculated into liquid culture medium and cultured with shaking under preset conditions for 24-36 hours to obtain the first bacterial culture.

[0014] The *Pseudomonas atagonensis* FJS-16 was inoculated into liquid culture medium and cultured under the preset conditions with shaking for 24-36 hours to obtain a second bacterial culture.

[0015] The first bacterial solution and the second bacterial solution are mixed at a preset volume ratio to obtain the composite bacterial agent.

[0016] Furthermore, the preset conditions include: a culture temperature of 28-30℃ and an oscillation speed of 180-220 rpm during the culture process.

[0017] Furthermore, the preset volume ratio is 1:1.

[0018] The present invention also provides a solid microbial fertilizer, comprising: a compound microbial agent as described in any of the above claims; wherein the compound microbial agent is derived from the compound microbial agent as described in any of the above claims.

[0019] Furthermore, the solid microbial fertilizer also includes: peat soil, physiological saline, and a 1% sodium alginate solution.

[0020] This invention also provides a method for preparing solid microbial fertilizer as described in any one of the above claims, comprising: separating microbial cells from the compound microbial agent as described in any one of the above claims; and then resuspending the microbial cells in the physiological saline solution to obtain OD. 600 Resuspension of bacterial cells with a value of 0.5;

[0021] The resuspended bacterial cells and the sodium alginate solution were mixed at a volume ratio of 2:1 to obtain the treatment solution.

[0022] The treatment solution and the peat soil are mixed at a volume ratio of 4:5 to obtain the solid microbial fertilizer.

[0023] The present invention also provides the application of the compound bacteria as described in any one of the above claims in promoting the growth of tea trees and / or tobacco.

[0024] Compared with the prior art, the present invention has the following advantages:

[0025] The growth-promoting compound bacteria provided by this invention include Burkholderia pyrrocinia FJS-3 and Pseudomonas atagonensis FJS-16. Studies have shown that both Burkholderia pyrrocinia FJS-3 and Pseudomonas atagonensis FJS-16 possess phosphate-solubilizing and potassium-solubilizing functions, and both can produce IAA, NH3, heptaphilin, and ACC deaminase, thus exhibiting superior growth-promoting effects. Furthermore, although Burkholderia pyrrocinia FJS-3 has nitrogen-fixing capabilities, the growth-promoting effect of Pseudomonas atagonensis FJS-16 is more pronounced.

[0026] When the growth-promoting compound bacteria of this invention are used as solid microbial fertilizer, the microbial fertilizer obtained by this invention can not only promote the growth of tea trees, increase the yield of tea trees and the quality of tea, and reduce the use of chemical fertilizers, thus achieving the goal of reducing fertilizer use and increasing yield in tea trees; moreover, the microbial fertilizer obtained by this invention also has a certain promoting effect on the growth of tobacco. Attached Figure Description

[0027] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the structures shown in these drawings without creative effort.

[0028] Figure 1 This is a colony morphology diagram of FJS-3 in Example 1;

[0029] Figure 2 Gram staining image of FJS-3 in Example 1;

[0030] Figure 3 This is a colony morphology diagram of FJS-16 in Example 1;

[0031] Figure 4 Gram staining image of FJS-16 in Example 1;

[0032] Figure 5 This is a growth status diagram of the microbial fertilizer treatment group in Example 5;

[0033] Figure 6 This is a growth status diagram of the control group in Example 5;

[0034] Figure 7 This is a growth state diagram of treatment 1 in Example 7;

[0035] Figure 8 This is a growth state diagram of treatment 2 in Example 7;

[0036] Figure 9 This is a growth state diagram of treatment 3 in Example 7;

[0037] Figure 10 This is a growth state diagram of treatment 4 in Example 7;

[0038] Figure 11 The diagram shows the root growth status of each treatment group in Example 8;

[0039] Among them, (a) is the root growth status diagram corresponding to treatment 1; (b) is the root growth status diagram corresponding to treatment 2; (c) is the root growth status diagram corresponding to treatment 3; and (d) is the root growth status diagram corresponding to treatment 4.

[0040] Figure 12 This is a growth status diagram of each treatment group in Example 8.

[0041] The realization of the objective, functional characteristics and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0042] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0043] The technical solutions of the various embodiments of the present invention can be combined with each other, but this must be based on the ability of those skilled in the art to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be considered that such combination of technical solutions does not exist and is not within the scope of protection claimed by the present invention.

[0044] It is worth noting that microbial fertilizers made from plant growth-promoting rhizobacteria (PGPR) have received widespread acclaim for their environmental friendliness, resource conservation, and green safety.

[0045] Studies have shown that microbial fertilizers play an irreplaceable role in soil repair, soil fertility improvement, fertilizer utilization, crop disease incidence reduction, and crop quality enhancement.

[0046] In view of this, the present invention provides a growth-promoting bacterium and a growth-promoting compound bacterium. The growth-promoting bacterium can be called a rhizosphere PGPR growth-promoting bacterium, and the growth-promoting compound bacterium can be called a rhizosphere PGPR growth-promoting compound bacterium. Since the growth-promoting bacterium and the growth-promoting compound bacterium have superior growth-promoting effects on tea trees and tobacco, the growth-promoting bacterium can also be called a tea tree / tobacco rhizosphere PGPR growth-promoting bacterium, and the growth-promoting compound bacterium can also be called a tea tree / tobacco rhizosphere PGPR growth-promoting compound bacterium.

[0047] Specifically, the growth-promoting bacteria are either Burkholderia pyrogallol or Pseudomonas atagonii. The growth-promoting complex includes or may include Burkholderia pyrogallol and Pseudomonas atagonii.

[0048] The *Burkholderia pyrrocinia* described is *Burkholderia pyrrocinia* FJS-3, with the Latin taxonomic name *Burkholderia pyrrocinia*. It has been deposited at the Guangdong Provincial Microbial Culture Collection Center on July 20, 2022, with accession number GDMCC No. 62432. *Burkholderia pyrrocinia* FJS-3 is a Gram-negative bacterium, and its colony morphology consists of pale yellow, moist, round dots.

[0049] The *Pseudomonas atagonensis* described is *Pseudomonas atagonensis* FJS-16, with the Latin taxonomic name *Pseudomonas atagonensis*. It has been deposited at the Guangdong Provincial Microbial Culture Collection Center on July 20, 2022, with accession number GDMCC No. 62433. *Pseudomonas atagonensis* FJS-16 is a Gram-negative bacterium, and its colony morphology consists of moist, milky-white dots.

[0050] Based on the above-mentioned growth-promoting compound bacteria, the present invention also provides a compound bacterial agent, including the compound bacteria as described in any of the above embodiments.

[0051] The preparation process of the compound microbial agent may specifically include:

[0052] First, Burkholderia pyrrocinia FJS-3 and Pseudomonas atagonensis FJS-16 were inoculated into separate LB solid medium and cultured overnight.

[0053] Then, single colonies of the two strains were picked from the corresponding LB solid medium and added to the corresponding LB liquid medium of the two strains respectively, and cultured under preset conditions until the logarithmic growth phase.

[0054] Subsequently, Burkholderia pyrrocinia FJS-3 was inoculated into LB liquid medium at a volume ratio of 5%, and cultured with shaking under the preset conditions for 24-36 hours to obtain the first bacterial culture.

[0055] The *Pseudomonas atagonensis* FJS-16 was inoculated into LB liquid medium at a volume ratio of 5% and cultured under the preset conditions with shaking for 24-36 hours to obtain a second bacterial culture.

[0056] Finally, the first bacterial solution and the second bacterial solution are mixed at a preset volume ratio to obtain the composite bacterial agent.

[0057] Furthermore, the LB solid culture medium contains 10.0g of tryptone, 10.0g of sodium chloride, 5.0g of yeast extract, 1000ml of distilled water, and 18g of agar; the pH of the solid culture medium is 7.0-7.2, and it is sterilized at 121℃ for 20min before use.

[0058] The LB liquid culture medium contains 10.0g tryptone, 10.0g sodium chloride, 5.0g yeast extract, and 1000ml distilled water; the solid culture medium has a pH of 7.0-7.2 and is sterilized at 121℃ for 20min before use.

[0059] Furthermore, the preset conditions for culturing Burkholderia pyrrocinia FJS-3 and Pseudomonas atagonensis FJS-16 can be: a culture temperature of 28-30℃ and an oscillation speed of 180-220 rpm during the culture process.

[0060] Furthermore, in the preparation of the compound bacterial agent, the first bacterial solution and the second bacterial solution can be mixed in equal proportions, that is, the preset volume ratio can be 1:1.

[0061] To meet actual planting needs, the present invention also provides a solid microbial fertilizer for promoting plant production and improving plant quality, comprising: a compound microbial agent as described in any of the above embodiments, wherein the compound microbial agent may be derived from the compound microbial agent described in any of the above embodiments.

[0062] It also includes: sterilized peat moss, sterile physiological saline (0.9%), and a 1% sterile sodium alginate solution (0.01g / ml).

[0063] To facilitate those skilled in the art in obtaining the solid microbial fertilizer, the present invention also provides a method for preparing the solid microbial fertilizer as described in any of the above embodiments, comprising:

[0064] After centrifuging the compound bacterial agent, the centrifuged (separated) bacterial cells are resuspended in 0.9% physiological saline and the OD is adjusted. 600 A value of 0.5 was used to prepare resuspended bacterial cells;

[0065] A 1% sodium alginate solution was added to the resuspended bacterial cells at a volume ratio of 2:1 and mixed thoroughly to obtain a treatment solution. The treatment solution was then mixed with sterilized peat at a volume ratio of 4:5 to obtain a solid bacterial fertilizer.

[0066] Since the compound bacteria in this invention have the effects of promoting growth, improving quality, and increasing yield of tea trees and tobacco, this method also provides an application of the compound bacteria as described in any of the above embodiments in promoting the growth of tea trees and / or tobacco.

[0067] Specifically, in practical applications, the compound bacteria can be prepared into the solid microbial fertilizer of the present invention; preferably, the solid microbial fertilizer and the compound fertilizer can be mixed and applied, for example, mixed and applied with the compound fertilizer in Example 6 of the present invention.

[0068] To facilitate a further understanding of the present invention by those skilled in the art, the following examples are provided:

[0069] Example 1

[0070] Strain screening:

[0071] 1. Four samples of rhizosphere soil from a tea garden in Tongren City, Guizhou Province were collected, placed in specimen bags, and stored in a refrigerator at 4°C for later use.

[0072] 2. Accurately weigh 10.0g of soil into each soil sample, add it into a 250mL Erlenmeyer flask containing glass beads and 90mL of sterile water, shake for 10min, and then let it stand.

[0073] 3. Take 1 mL of the supernatant from each soil sample, and then serially dilute it 10-fold to obtain a final concentration of 10. -2 10 -3 10 -4 10 -5 10 -6 Soil suspension.

[0074] 4. Using the spread method, the soil suspensions of the above concentrations were spread onto LB solid medium and PDA solid medium respectively. Colonies of different colors and different morphologies were picked out for purification and preservation.

[0075] 5. The purified strains were cultured and screened using potassium-solubilizing solid medium, phosphorus-solubilizing solid medium, and Ashby nitrogen-free solid medium.

[0076] Specifically, the purified strains were first cultured in potassium-solubilizing medium. Strains with a hydrolysis zone diameter / colony diameter ratio > 2.0 were placed on the other two selective media for further culture to obtain strains with nitrogen fixation, phosphorus solubilization, and potassium solubilization functions (denoted as FJS-3) and strains with phosphorus solubilization and potassium solubilization functions (denoted as FJS-16).

[0077] Among them, the colony morphology of strain FJS-3 on LB medium is as follows: Figure 1 As shown, its colonies appear as pale yellow, moist dots; the Gram staining image of strain FJS-3 is shown below. Figure 2 As shown, it is a Gram-negative bacterium.

[0078] The colony morphology of strain FJS-16 on LB medium is as follows: Figure 3 As shown, its colonies appear as milky white, moist dots. The Gram staining image of strain FJS-16 is shown below. Figure 4 As shown, it is a Gram-negative bacterium.

[0079] The culture medium involved in this embodiment is as follows:

[0080] LB solid medium: 10.0g tryptone, 10.0g sodium chloride, 5.0g yeast extract, 1000ml distilled water, 18g agar; adjust pH to 7.0.

[0081] PDA solid culture medium: 200g potato, 20g glucose, 15g agar, 1000ml distilled water, natural pH.

[0082] Potassium-solubilizing solid culture medium: sucrose 5g, Na2HPO4 2g, MgSO4·7H2O 0.5g, FeCl3 0.005g, CaCO3 0.1g, potassium feldspar powder 1.0g, agar 15g, distilled water 1000ml, pH 7.2.

[0083] Phosphate-solubilizing solid culture medium: glucose 10.0g, ammonium sulfate 0.5g, sodium chloride 0.3g, magnesium sulfate 0.3g, manganese sulfate 0.03g, ferrous sulfate 0.03g, calcium phosphate 5.0g, yeast extract 0.5g, potassium chloride 0.3g, agar 15.0g, pH 7.0.

[0084] Ashby's nitrogen-free solid medium: mannitol 10g, KH2PO4 0.2g, MgSO4·7H2O 0.2g, NaCl 0.2g, CaSO4·2H2O 0.1g, CaCO3 5g, agar 15.0g, distilled water 1000ml, pH 7.0.

[0085] Example 2

[0086] Identification of strains

[0087] Both FJS-3 and FJS-16 strains from Example 1 underwent 16S rDNA amplification and molecular identification:

[0088] The methods for 16S rDNA gene amplification and molecular identification are as follows:

[0089] 1. Total DNA was extracted from the bacterial strain using a bacterial DNA extraction kit. Using the total bacterial DNA as a template, PCR amplification was performed using universal primers for the prokaryotic 16S rDNA gene.

[0090] 2. Sequencing the amplified products;

[0091] 3. The obtained sequences were compared with those from the NCBI and EzTaxon databases using BLAST analysis, and the 16S rDNA gene sequences of the type strains with high homology were selected.

[0092] The 16S rDNA sequence of strain FJS-3 is shown below:

[0093]

[0094] The 16S rDNA sequence of strain FJS-16 is shown below:

[0095]

[0096] Analysis revealed that the FJS-3 strain screened in Example 1 was Burkholderia pyrrocinia, which is Burkholderia pyrrocinia in this invention. This invention names it Burkholderia pyrrocinia FJS-3.

[0097] The FJS-16 strain screened in Example 1 is Pseudomonas atagonensis, which is the Pseudomonas atagonensis in this invention. This invention names it Pseudomonas atagonensis FJS-16.

[0098] Example 3

[0099] Study on the growth-promoting effect of strains

[0100] 1. IAA secretion capacity assay

[0101] The Salkowski method was used to determine the ability of the test strain to produce indoleacetic acid (IAA). A 1 mL tube of the test strain, preserved in glycerol, was inoculated into 50 mL of LB broth containing 100 mg / L tryptophan (filtered and sterilized) and incubated at 30°C and 180 rpm for 1 day. 50 μL of the bacterial suspension was dropped onto a white ceramic plate, and an equal volume of Salkowski colorimetric solution was added; a colorimetric solution containing 50 mg / L indoleacetic acid was used as a positive control. The white ceramic plate was incubated at room temperature in the dark for 30 minutes. A reddish color indicated that the strain could secrete IAA.

[0102] 2. NH3 production capacity determination

[0103] The test strain preserved in a 100 μL glycerol tube was inoculated into a test tube containing 10 mL of peptone water (10 g / L) and cultured at 28 °C for 3 days. Then, 0.5 mL of Nessler's reagent was added to each tube. The color change from brown to yellow indicates that NH3 was produced.

[0104] 3. Determination of the ability to produce ferrophiles

[0105] The ability to produce ferophiles was determined using the CAS method. The test strain, preserved in 100 μL glycerol tubes, was inoculated onto a CAS test plate and cultured at 28°C for 3 days. The appearance of a yellow-green halo around the strain indicated that the strain had the ability to produce ferophiles.

[0106] 4. Qualitative determination of ACC deaminase activity

[0107] The test strain preserved in 50 μL glycerol tubes was inoculated into 5 mL of nitrogen-free liquid medium and cultured at 30°C with shaking at 200 rpm for 1 day; 0.1 mL of the culture solution was then inoculated into 5 mL of DF medium and cultured with shaking for 1 day; finally, 2% of the culture solution was transferred to 5 mL of ALADF medium and cultured for another 2 days. Strains that can grow using ACC as the sole nitrogen source are ACC deaminase-positive strains.

[0108] The functions of strains FJS-3 and FJS-16 are as follows:

[0109]

[0110] Note: "+" indicates that the bacteria have this function, and "-" indicates that the bacteria do not have this function.

[0111] Example 4

[0112] 1. The preparation process of the solid microbial fertilizer (rhizosphere PGPR growth-promoting microbial fertilizer) in this implementation is as follows:

[0113] After centrifuging the compound bacterial agent, resuspend it in the 0.9% sterile physiological saline solution and adjust the OD value. 600 A value of 0.5 was used to prepare resuspended bacterial cells;

[0114] A treatment solution was prepared by adding 1% sterile sodium alginate solution to the resuspended bacterial cells at a volume ratio of 2:1 and mixing thoroughly. The treatment solution was then mixed with sterilized peat at a volume ratio of 4:5 to obtain solid bacterial fertilizer.

[0115] The number of viable bacteria fixed in the solid microbial fertilizer was 13.48±0.16 LgN (cfu / g), and the number of viable bacteria released after 60 days of storage at room temperature was 8.46±0.14 LgN (cfu / g).

[0116] The solid microbial fertilizers used in the subsequent embodiments of the present invention are all freshly prepared solid microbial fertilizers.

[0117] 2. The preparation process of the compound microbial agent is as follows:

[0118] FJS-3 and FJS-16 strains were inoculated into their respective LB solid medium and cultured overnight.

[0119] Liquid activation: A single colony was picked from LB solid medium inoculated with FJS-3 strain and placed into LB liquid medium. The culture was carried out at 28°C and shaken at 220 rpm until the logarithmic growth phase. This was recorded as liquid a.

[0120] Single colonies were picked from LB solid medium inoculated with FJS-16 strain and placed into LB liquid medium. The culture was carried out at 28°C and shaken at 220 rpm until the logarithmic growth phase. This was recorded as solution b.

[0121] Fermentation broth of each strain was obtained: 5% by volume of strain a in the logarithmic growth phase was inoculated into LB liquid medium and cultured at 28℃ with shaking at 220 rpm for 24 h to obtain FJS-3 fermentation broth.

[0122] The b solution, which was in the logarithmic growth phase, was inoculated into LB liquid medium at a volume of 5% and cultured at 28°C with shaking at 220 rpm for 24 h to obtain FJS-16 fermentation broth.

[0123] Fermentation broth mixing: Mix equal volumes of FJS-3 fermentation broth and FJS-16 fermentation broth evenly to obtain a compound microbial agent.

[0124] Example 5

[0125] Laboratory potted application for promoting tea seedling growth

[0126] Experimental crop: Tea seedlings

[0127] Experimental variety: Fuding Dabai

[0128] The experiment was designed with two treatment groups, each with three replicates: the microbial fertilizer treatment group (applied solid microbial fertilizer as described in Example 4) and the control group (no fertilizer applied).

[0129] Tea seedlings of similar size and growth were selected for transplanting, and the water management and the weight of the nutrient soil in the transplanting pots were kept consistent for each group.

[0130] Application method of microbial fertilizer: Apply 1g / pot 10 days and 20 days after transplanting tea seedlings.

[0131] Photos were taken and recorded 30 days after transplanting, and the biochemical components of the tea leaves were measured.

[0132] In this embodiment, the growth of tea seedlings in the microbial fertilizer treatment group is as follows: Figure 5 As shown; the growth of tea seedlings in the control group is as follows. Figure 6 As shown in the figure, the tea seedlings in the microbial fertilizer treatment group grew significantly better than those in the control group.

[0133] The content of biochemical components in tea leaves in this embodiment is shown in the table below:

[0134]

[0135] It should be noted that chlorophyll is an important component of the color of tea soup and the color of dry tea leaves; tea polyphenols and caffeine are important components of the astringency in tea soup; and amino acids in tea leaves are one of the important metabolic products in the nitrogen cycle, and also one of the important components of the fresh and refreshing taste in tea soup.

[0136] The table above shows that the chlorophyll content in the microbial fertilizer treatment group was higher than that in the control group; the tea polyphenol and caffeine content in the control group was higher than that in the microbial fertilizer treatment group; and the amino acid content in the microbial fertilizer treatment group was higher than that in the control group. Therefore, it can be inferred that the tea quality in the microbial fertilizer treatment group was superior to that in the control group.

[0137] Example 6

[0138] Field applications for increasing tea yield and improving quality

[0139] The experimental site was located in Guanyin Mountain Tea Garden, Jiangkou County, Tongren City, Guizhou Province (E108°47′52″, N27°41′24″), at an altitude of 780m. The tea variety was Longjing 43, and the tea plantations in the garden were about 10-15 years old.

[0140] The experiment was set up with 3 treatment groups, each treatment was repeated three times, and each repeat occupied 0.3 acres.

[0141] The experiment was conducted from November 2021 to April 2022. All fertilizers were applied evenly to each plot before the tea trees entered winter (November 2021), and no further topdressing was applied afterward. Except for the source of fertilizers, the other treatment conditions of the three treatment groups were the same.

[0142] Treatment 1: No fertilizer applied (CK);

[0143] Treatment 2: The solid microbial fertilizer and compound fertilizer from Example 4 were mixed and applied together, with the solid microbial fertilizer applied at a rate of 2 g / m³. 2 The application rate of compound fertilizer is 0.25 kg / m³. 2 ;

[0144] Treatment 3: Apply the solid microbial fertilizer from Example 4 at a rate of 2 g / m³. 2 ;

[0145] Among them, the compound fertilizer has a total nutrient content (N+P2O5+K2O) ≥6%, organic matter ≥60%, (boron+magnesium+zinc+calcium+sulfur) ≥3.6%, and a total number of beneficial microbial colonies (Bacillus subtilis, Bacillus mucilaginosa, photosynthetic bacteria, etc.) ≥0.3 billion / gram.

[0146] The following are the experimental results from the 6th month after fertilization (April 2022):

[0147] 1. The tea yield of each treatment group is compared in the table below.

[0148]

[0149] 2. The amino acid content of tea leaves in each treatment group is shown in the table below:

[0150]

[0151]

[0152] 3. The polyphenol content in tea leaves of each treatment group is shown in the table below:

[0153]

[0154] 4. The caffeine content in the tea leaves of each treatment group is shown in the table below:

[0155]

[0156] 5. The germination density comparison of each treatment group is shown in the table below. It can be seen that:

[0157]

[0158] 6. The comparison of the quality of 100 shoots in each treatment group is shown in the table below:

[0159]

[0160] Example 7

[0161] Laboratory potted application for promoting tea seedling growth

[0162] Experimental crop: Tea seedlings

[0163] Experimental variety: Fuding Dabai

[0164] The experiment was designed with 4 treatment groups, each with 3 replicates:

[0165] Tea seedlings of similar size and growth were selected for transplanting, and the water management and the weight of the nutrient soil in the transplanting pots were kept consistent for each treatment group.

[0166] Treatment 1 (CK): No fertilizer applied;

[0167] Treatment 2: Apply solid microbial fertilizer containing FJS-3 bacteria (compared to the solid microbial fertilizer in Example 4, only the source of microorganisms was replaced from compound microbial agents to microbial agents of FJS-3 strain, and the OD of the resuspended bacteria was measured). 600 (Other preparation conditions, such as the value, remain unchanged);

[0168] Treatment 3: Apply solid microbial fertilizer containing FJS-16 bacteria (compared to the solid microbial fertilizer in Example 4, only the source of microorganisms was replaced from compound microbial agents to microbial agents containing FJS-16 strains, and the OD of the resuspended bacteria was measured). 600 (Other preparation conditions, such as the value, remain unchanged);

[0169] Treatment 4: Apply the solid microbial fertilizer from Example 4.

[0170] Application method: Apply 1g / pot 10 days and 20 days after transplanting tea seedlings.

[0171] Experimental results:

[0172] At 30 days after transplanting, the growth of tea seedlings in the control group (treatment 1) was as follows: Figure 7 As shown, the growth of tea seedlings after FJS-3 (treatment 2) treatment is as follows: Figure 8 As shown, the growth of tea seedlings after FJS-16 (treatment 3) treatment is as follows: Figure 9 As shown, the growth of tea seedlings after treatment with compound microbial agent (treatment 4) is as follows. Figure 10 As shown.

[0173] It can be seen that in different treatment groups, the number of tea buds sprouting and the length of new shoots were all in the order of compound microbial agent > FJS-16 > FJS-3 > control group. This indicates that both FJS-3 and FJS-16, as single microbial agents, can promote tea seedling growth, but the growth-promoting effect of the compound microbial agent is significantly better than that of the single microbial agent. Moreover, observations show that microbial fertilizer mainly promotes the growth and yield of tea trees by promoting the sprouting of tea buds and the growth of branches.

[0174] Example 8

[0175] Laboratory potted plant application for tobacco growth promotion

[0176] Experimental crop: Tobacco

[0177] Test variety: K326

[0178] In this embodiment, a total of 4 processing groups are designed, and each processing group has 3 repetitions.

[0179] Tobacco was bred using the soil-sowing method. When the tobacco plants had grown to the fourth true leaf, they were transplanted. The water management, the weight of the nutrient soil in the transplanting pots, and other conditions were kept consistent across all treatment groups.

[0180] Treatment 1 (CK): No fertilizer applied;

[0181] Treatment 2 (FIS-3): Apply solid microbial fertilizer containing FJS-3 bacteria; (Compared to the solid microbial fertilizer in Example 4, only the source of microorganisms was replaced from compound microbial agents to microbial agents of FJS-3 strain, and the OD of the resuspended bacteria was...) 600 (Other preparation conditions, such as the value, remain unchanged);

[0182] Treatment 3 (FIS-16): Apply solid microbial fertilizer containing FJS-16 bacteria (compared to the solid microbial fertilizer in Example 4, only the source of microorganisms was replaced from the compound microbial agent to the FJS-16 strain microbial agent, and the OD of the resuspended bacteria was measured). 600 (Other preparation conditions, such as the value, remain unchanged);

[0183] Treatment 4 (FIS-3+16): Apply the solid microbial fertilizer from Example 4.

[0184] Application method: Apply 1g / pot 10 days and 20 days after tobacco transplanting.

[0185] Experimental results:

[0186] 1. The tobacco plant height 30 days after transplanting is shown in the table below:

[0187]

[0188] It can be seen that, in terms of the effect on plant height, compound microbial agent > FJS-16 > FJS-3 > CK, indicating that the growth-promoting effect of compound microbial agent is better than that of single microbial agent.

[0189] 2. The fresh weight of tobacco 30 days after transplanting is shown in the table below:

[0190]

[0191] It can be seen that, in terms of effect on fresh weight, compound microbial agent > FJS-16 > FJS-3 > CK, indicating that the growth-promoting effect of compound microbial agent is better than that of single microbial agent.

[0192] 3. The root weight of the tobacco plants 30 days after transplanting is shown in the table below:

[0193]

[0194] It can be seen that, in terms of the effect on plant height, compound microbial agent > FJS-16 > FJS-3 > CK, indicating that the growth-promoting effect of compound microbial agent is better than that of single microbial agent.

[0195] 4. The stem diameter of the tobacco plant 30 days after transplanting is shown in the table below:

[0196]

[0197] It can be seen that, in terms of the effect on stem diameter, the compound microbial agent > FJS-16 > FJS-3 > CK, indicating that the growth-promoting effect of the compound microbial agent is better than that of the single microbial agent.

[0198] 5. The root growth status of tobacco plants 30 days after transplanting is as follows: Figure 11 As shown, in terms of the effect on root development, compound microbial agent > FJS-16 > FJS-3 > CK, indicating that the growth-promoting effect of compound microbial agent is better than that of single microbial agent.

[0199] 6. The growth status of tobacco 30 days after transplanting is as follows: Figure 12As shown, in terms of the effect on growth status, compound microbial agent > FJS-16 > FJS-3 > CK.

[0200] The above technical solutions of the present invention are merely preferred embodiments of the present invention and do not limit the patent scope of the present invention. All equivalent structural transformations made under the technical concept of the present invention using the contents of the present invention specification and drawings, or direct / indirect applications in other related technical fields, are included in the patent protection scope of the present invention.

Claims

1. A growth-promoting compound bacteria, characterized in that, The growth-promoting compound bacteria include Burkholderia pyrrole and Pseudomonas aeruginosa. The *Burkholderia pyrogallae* is *Burkholderia pyrogallae* (… Burkholderia pyrrocinia FJS-3, named in Latin classification Burkholderia pyrrocinia It has been deposited at the Guangdong Provincial Center for Microbial Culture Collection on July 20, 2022, with accession number GDMCC No. 62432; The Atagonian Pseudomonas is Atagonian Pseudomonas ( Pseudomonas atagonensis FJS-16, named in Latin classification Pseudomonas atagonensis It has been deposited at the Guangdong Provincial Center for Microbial Culture Collection on July 20, 2022, with accession number GDMCC No. 62433.

2. A compound microbial agent, characterized in that, Includes the growth-promoting compound bacteria as described in claim 1.

3. The compound microbial agent according to claim 2, characterized in that, The preparation process of the compound microbial agent includes: The aforementioned Burkholderia pyrrole ( Burkholderia pyrrocinia FJS-3 was inoculated into liquid culture medium and cultured with shaking under preset conditions for 24-36 hours to obtain the first bacterial culture. The Atagonian Pseudomonas ( Pseudomonas atagonensis FJS-16 was inoculated into liquid culture medium and cultured with shaking under the preset conditions for 24-36 hours to obtain the second bacterial culture. The first bacterial solution and the second bacterial solution are mixed at a preset volume ratio to obtain the composite bacterial agent.

4. The compound microbial agent according to claim 3, characterized in that, The preset conditions include: a culture temperature of 28-30℃ and an oscillation speed of 180-220 rpm during the culture process.

5. The compound microbial agent according to claim 3, characterized in that, The preset volume ratio is 1:

1.

6. A solid microbial fertilizer, characterized in that, include: The growth-promoting compound bacteria as described in claim 1; wherein the growth-promoting compound bacteria are derived from the compound bacterial agent as described in any one of claims 2-5.

7. The solid microbial fertilizer according to claim 6, characterized in that, The solid microbial fertilizer also includes: peat soil, physiological saline, and a 1% sodium alginate solution.

8. The application of the growth-promoting compound bacteria as described in claim 1 in promoting the growth of tea trees and / or tobacco.

Images