Burkholderia gladiolus and its bacterial agent and application

Abstract

The present invention relates to the field of microbial technology, and discloses a gladiolus Burkholderia and its bacterial agent and application. The gladiolus Burkholderia provided by the present invention ( Burkholderia gladioli )CCTCC NO: M20242288 has good nitrogen fixation, phosphorus solubilization, zinc solubilization, iron carrier production and sulfur oxidation capabilities, can adjust the soil nutrient structure, transform the soil substances that are difficult for plants to use, thereby promoting plant growth and improving plant stress resistance; this strain can also antagonize Fusarium solani ( Fusarium nightshade ), Fusarium equisetum ( Fusarium horsetail )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ); In addition, the gladiolus Burkholderia provided by the present invention is a strain isolated from rapeseed farmland soil. Compared with strains screened by extreme environments and laboratory mutagenesis, it has better adaptability to planting environments, can stably colonize in the rhizosphere and soil of plants, thereby playing a long-term and stable growth-promoting role, is safe and environmentally friendly, and has broad application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a gladiolus Burkholderia and a bacterial agent and application thereof. Background Art

[0002] With the rapid development of modern agriculture, the concept of sustainable development has become increasingly popular. However, soil, as the basic resource for agricultural production, is facing unprecedented challenges. Long-term over-fertilization, abuse of pesticides and unreasonable farming methods have led to the destruction of soil structure, the decline of fertility, and even serious soil pollution problems, which have seriously restricted the sustainable development of agriculture. Therefore, it is particularly urgent to find a green and environmentally friendly measure that can effectively improve soil conditions.

[0003] Microorganisms are an indispensable part of nature and a valuable natural resource. They play an important role in agricultural development. They can promote plant growth, enhance plant resistance to diseases, improve soil structure and increase nutrient circulation in the soil. In particular, plant growth promoting rhizobacteria (PGPR) can colonize around plant roots and directly or indirectly promote plant growth and increase crop yields by fixing nitrogen, solubilizing phosphorus and producing growth hormones. In addition, PGPR can inhibit the growth of pathogenic microorganisms and reduce the occurrence of plant diseases. They can form a mutually beneficial symbiotic relationship with plants, reduce the use of chemical fertilizers and pesticides, reduce production costs, protect the environment and achieve green and sustainable development of agriculture.

[0004] Although plant rhizosphere growth-promoting bacteria have broad application prospects in agriculture, the current research on these microorganisms is still in the stage of continuous exploration and development. The existing rhizosphere growth-promoting bacteria are diverse in species and have different functions, but not all strains can show good growth-promoting effects under all soil and crop conditions. Therefore, in-depth exploration and screening of more PGPR strains with efficient colonization ability and wide functions will not only help develop new biofertilizer products and improve agricultural production efficiency, but also further promote agriculture to develop in a more green, environmentally friendly and sustainable direction. Summary of the invention

[0005] The present invention aims to solve the problem that the prior art lacks effective plant growth-promoting bacteria and their products in practical applications, and provides gladiolus Burkholderia and its bacterial agent and application. The gladiolus Burkholderia provided by the present invention is isolated from rapeseed farmland soil in Dali City, Yunnan Province. The bacterium has the characteristics of sulfur oxidation, nitrogen fixation, zinc solubilization, organic phosphorus and inorganic phosphorus solubilization, and iron carrier production, and can also antagonize Fusarium solani ( Fusarium solani ), Fusarium equisetum ( Fusariumequiseti )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ).

[0006] To achieve the above object, the present invention provides a strain of Burkholderia gladiolus ( Burkholderia gladioli YNK), the deposit number of this strain is CCTCC NO: M20242288.

[0007] The second aspect of the present invention provides a bacterial agent, the active ingredient of which is the Burkholderia gladiolus of the first aspect.

[0008] The third aspect of the present invention provides use of the Burkholderia gladiolus described in the first aspect or the bacterial agent described in the second aspect in sulfur oxidation and zinc dissolution.

[0009] The fourth aspect of the present invention provides the gladiolus Burkholderia described in the first aspect in antagonizing Fusarium solani ( Fusarium solani ), Fusarium equisetum ( Fusarium equiseti )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ) in applications.

[0010] Through the above technical solution, the present invention can at least achieve the following beneficial effects:

[0011] (1) The gladiolus Burkholderia provided by the present invention has excellent sulfur oxidation, nitrogen fixation, zinc solubilization, organic phosphorus and inorganic phosphorus solubilization, and siderophore production functions, and can also antagonize Fusarium solani ( Fusarium solani ), Fusarium equisetum ( Fusarium equiseti )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ), which can effectively prevent and control diseases, improve soil quality, and increase the content of nutrients that can be used by plants in the soil, thereby having a good effect in promoting plant growth.

[0012] (2) The gladiolus Burkholderia provided by the present invention is a strain isolated from rapeseed farmland soil. Compared with strains screened in extreme environments and laboratory mutagenesis, it has better adaptability to the planting environment and can stably colonize in the rhizosphere and soil of plants, thereby playing a long-term and stable growth-promoting role. Moreover, since it is a strain isolated from farmland soil, the risk of the strain causing adverse effects on the local environment and microbial ecology in the soil during its application in farmland is reduced. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 1 is a colony morphology image (A) and an electron microscope scanning image (B) of Burkholderia gladiolus in Example 1;

[0014] Figure 2 is the phylogenetic tree of Burkholderia gladiolus in Example 1;

[0015] Figure 3 The diagrams are the effects of promoting nutrient cycle of gladiolus Burkholderia in Example 2: (A) sulfur oxidation effect diagram; (B) nitrogen fixation effect diagram; (C) zinc dissolution effect diagram; (D) organic phosphorus decomposition effect diagram; (E) inorganic phosphorus decomposition effect diagram; (F) siderophore production effect diagram;

[0016] Figure 4 is a standard curve diagram drawn in the quantification of sulfur oxidation effect of Burkholderia gladiolus in Example 2;

[0017] Figure 5 The antagonistic effect diagram of strain YNK-FB0053 in Example 3 is as follows: (A) Antagonism against Fusarium solani ( Fusarium solani ) Effect diagram; (B) Antagonism against Fusarium equisetum ( Fusarium equiseti ) Effect diagram; (C) Antagonism against Fusarium rotundifolia ( Fusarium concentricum ) effect diagram; (D) antagonism to brown seat crust fungus ( Rosellinia necatrix ) effect diagram.

[0018] Biological Deposit

[0019] The gladiolus Burkholderia provided by the present invention is classified and named Burkholderia gladioli YNK-FB0053 was deposited in the China Center for Type Culture Collection on October 21, 2024, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, Wuhan University, with the deposit number CCTCC NO: M20242288. DETAILED DESCRIPTION

[0020] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0021] The inventor of the present invention accidentally isolated a gladiolus Burkholderia from rapeseed farmland soil in Dali City, Yunnan Province during the research process. It was found that the strain had good sulfur oxidation ability, as well as nitrogen fixation, zinc solubilization, organic and inorganic phosphorus solubilization, siderophore production, and antagonism to pathogens (including Fusarium solani ( Fusarium solani ), Fusarium equisetum (Fusarium equiseti )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix )) performance, the strain can not only promote nutrient circulation, but also antagonize pathogens. After further research, the inventors also found that it can effectively promote plant growth, improve agricultural productivity, and promote the green and sustainable development of ecological agriculture.

[0022] Based on the above findings, the present invention provides a strain of Burkholderia gladiolus. Burkholderia gladioli YNK-FB0053, the deposit number of this strain is CCTCC NO: M20242288.

[0023] The second aspect of the present invention provides the use of the gladiolus Burkholderia described in the first aspect in sulfur oxidation, nitrogen fixation, zinc dissolution, organic and inorganic phosphorus decomposition, and siderophore production.

[0024] In the present invention, "nitrogen fixation" and "phosphorus solubilization" refer to the increase of the N and P content in the soil that can be used by plants through the action of the gladiolus Burkholderia provided by the present invention. For example, the inorganic phosphorus components (such as Ca3(PO4) 2、 FePO4, etc.) and organic phosphorus components (such as lecithin, inositol hexaphosphate, etc.) are converted into a form that can be used by plants, thereby promoting the absorption and utilization of phosphorus in the soil by plants.

[0025] A third aspect of the present invention provides a method for sulfur oxidation, comprising culturing the Burkholderia gladiolus described in the first aspect, and collecting the culture product.

[0026] Any method and conditions that can ferment and culture Burkholderia gladiolus and cause it to oxidize sulfur can be applied to the present invention.

[0027] The fourth aspect of the present invention provides the gladiolus Burkholderia described in the first aspect in antagonizing Fusarium solani ( Fusarium solani ), Fusarium equisetum ( Fusarium equiseti )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ) in applications.

[0028] According to a preferred embodiment of the present invention, the fermentation culture conditions include: culture temperature 28-37°C, culture time 48-96h, initial pH = 6-8.

[0029] According to a preferred embodiment of the present invention, when the fermentation culture is carried out by means of shaking fermentation, the fermentation culture process can be carried out at a shaking speed of 120-200 rpm.

[0030] In the present invention, there is no particular limitation on the culture medium used for the fermentation culture, and any culture medium that can be used for fermentation culture of Burkholderia gladiolus in the art can be applied to the present invention.

[0031] The fifth aspect of the present invention provides a bacterial agent having the function of promoting plant growth, wherein the active ingredient in the bacterial agent includes the Burkholderia gladiolus described in the first aspect.

[0032] According to a preferred embodiment of the present invention, the bacterial agent is a liquid bacterial agent.

[0033] Preferably, the content of Burkholderia gladiolus in the liquid bacterial agent is not less than 1×10 6 CFU / mL.

[0034] More preferably, in the liquid bacterial agent, the content of Burkholderia gladiolus is 1×10 7 -1×10 10 CFU / mL. More preferably 1×10 8 -1×10 9 CFU / mL.

[0035] The present invention will be described in detail below by way of examples. It should be understood that the following examples are only used to further explain and illustrate the content of the present invention by way of example, and are not intended to limit the present invention.

[0036] In the following examples, unless otherwise specified, the reagents and materials used were all commercial products purchased from regular chemical / biological reagent or material suppliers, and the reagents were all analytical grade.

[0037] In the following examples, unless otherwise specified, the operating temperature is room temperature (25±5° C.).

[0038] Example 1

[0039] This example is used to illustrate the acquisition, identification and preservation of Burkholderia gladiolus CCTCC NO: M20242288.

[0040] (I) Strain isolation and purification

[0041] Thiosulfate agar medium (MST medium) was used in the strain isolation and purification process. The preparation method was as follows: weigh 5 g of sodium thiosulfate, 0.1 g of dipotassium hydrogen phosphate, 0.2 g of sodium bicarbonate, 0.1 g of ammonium chloride, 5 g of glucose, and 5 g of yeast extract powder, add them to 1000 mL of water, adjust the pH to 8±0.1, then add 20 g of agar, and sterilize at 121°C under high pressure for 25 min.

[0042] A bacterial strain named YNK-FB0053 was isolated and purified from rapeseed planting soil samples collected from Dali City, Yunnan Province by using the dilution spread plate method and the plate streak method.

[0043] (II) Strain identification

[0044] 1. Identification of bacterial morphological, physiological and biochemical characteristics

[0045] Physiological and biochemical tests were performed on strain YNK-FB0053 according to the Bergey's Manual of Bacterial Identification and the Manual of Identification of Common Bacterial Systems, and the colony morphology characteristics were described.

[0046] Colony and strain morphology: Figure 1 (A) shows the colony morphology of strain YNK-FB0053 on MST medium. It can be seen from the figure that the colony of this strain is nearly circular, with irregular edges, milky white, and a convex middle, moist and shiny. Figure 1 (B) This strain is rod-shaped and has no flagella.

[0047] Physiological and biochemical characteristics: Gram staining of strain YNK-FB0053 was negative, salt tolerance range was 0-5% (w / v), pH tolerance range was 5-9, indicating that the strain had certain salt and acid resistance. It was positive for sorbose, galactose, sorbitol, fructose, methyl red, catalase and oxidase, and hydrolysis of Tween 20, 40 and 80; it was negative for alkaline phosphatase, esterase (C4), starch hydrolase, tryptophan deaminase, sucrose, lactose, mannitol, maltose, soluble starch, inositol, and indole.

[0048] Molecular identification: The total DNA of strain YNK-FB0053 was extracted by Chelex extraction method as a template, 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) was used as the upstream primer, 1492R (5′-TACGGCTACCTTGTTACGACTT-3′) was used as the downstream primer, and 16S rRNA was amplified using the reaction system and conditions in Table 1.

[0049] Table 1 PCR system and conditions

[0050]

[0051]

[0052] Figure 2 The phylogenetic tree of strain YNK-FB0053 is shown, from which it can be seen that YNK-FB0053 is closely related to Burkholderia gladioli (Burkholderia gladioli )CFBP2427 has the highest homology.

[0053] 3. Identification results

[0054] Combined with the molecular detection results of strain YNK-FB0053 and the results of bacterial morphological and physiological and biochemical characteristics, the strain was identified as Burkholderia gladiolus ( Burkholderia gladioli ).

[0055] 3. Strain preservation

[0056] Name the above obtained classification as: Burkholderia gladioli The Burkholderia gladiolus of YNK-FB0053 was deposited in the China Center for Type Culture Collection, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, Wuhan University, on October 21, 2024, with the deposit number CCTCC NO: M20242288.

[0057] Example 2

[0058] This example is used to illustrate the sulfur oxidation, nitrogen fixation, zinc solubilization, organic and inorganic phosphorus solubilization, and siderophore production effects of Burkholderia gladioli CCTCC NO: M20242288.

[0059] (I) Sulfur oxidation effect test

[0060] 1. Qualitative testing

[0061] MST (modified thiosulfate) medium (g / L): weigh 5.0 g sodium thiosulfate, 0.1 g potassium dihydrogen phosphate, 0.2 g sodium bicarbonate, 0.1 g ammonium chloride, 5.0 g glucose, 5.0 g yeast extract powder, add to 1000 mL water to dissolve, add 0.008 g bromocresol purple as a color developer, adjust the pH to 8 ± 0.1, add 20 g agar, and sterilize at 121°C for 20 min.

[0062] The strain YNK-FB0053 obtained in Example 1 was inoculated into MST medium with a color developer by a 4-point inoculation method, and each treatment was repeated 3 times. The culture was placed in a 30°C incubator to continuously observe whether a yellow halo was generated around the colony. The ratio between the diameter (D) of the color development zone and the colony diameter (d) was measured.

[0063] Figure 3(A) shows the sulfur oxidation effect of strain YNK-FB0053. As can be seen from the figure, a yellow halo is produced in the culture medium around the colony, indicating that the strain is capable of sulfur oxidation. The size of the color circle is measured by the cross method, and the ratio of the color circle diameter (D) to the colony diameter (d) is calculated as D / d = 2.605 ± 0.272.

[0064] 2. Quantitative detection

[0065] MST liquid culture medium (g / L): Weigh 5.0 g of sodium thiosulfate, 0.1 g of dipotassium hydrogen phosphate, 0.2 g of sodium bicarbonate, 0.1 g of ammonium chloride, 5.0 g of glucose, and 5.0 g of yeast extract powder, add them to 1000 mL of water to dissolve, adjust the pH to 8±0.1, and sterilize by high pressure at 121°C for 20 min.

[0066] Preparation of stabilizing solution: Dissolve 75g analytical grade NaCl in 300ml water, add 30ml analytical grade concentrated HCl, 50ml glycerol and 100ml 95% ethanol, and mix well.

[0067] Preparation of sulfate standard stock solution: Weigh 1.814g of anhydrous potassium sulfate (K2SO4 analytical grade, dried at 105℃ for two hours in advance), dissolve it in a small amount of pure water, transfer it into a 1000ml volumetric flask, and dilute to the mark with pure water. 1.00ml of this solution contains 1.00mg sulfate (SO4 2- ).

[0068] Preparation of sulfate standard solution: Accurately pipette 10.00ml of the above stock solution into a 100ml volumetric flask and dilute to scale with pure water. 1.00ml of this solution contains 100ug sulfate (SO4 2- ).

[0069] Method for drawing a standard curve: weigh several portions of 0.1g BaCl2 crystals for later use, accurately add sulfate standard solution to a series of 50ml beakers, 0.00, 0.30, 0.50, 1.00, 1.25, 1.50, 2.00 and 2.50ml, add distilled water to a final volume of 5.00ml, then add 1ml of stabilizer respectively, place the 50ml beakers configured into a standard series on a magnetic stirrer one by one, quickly add 0.1g BaCl2 crystals while stirring, stir for 1min, let it stand for 15min, use a 2cm colorimetric dish to measure the absorbance at a wavelength of 420nm, use distilled water as a control, and draw a standard curve. The standard curve is shown below. Figure 4 shown.

[0070] Figure 4 The standard curve obtained by the above steps is y=0.0019x-0.0002, R²=0.9909, where x is SO42- content (mg / L), y is the corresponding absorbance value at this time.

[0071] The bacterial suspension was cultured in MST liquid medium, and after inoculation with gladiolus Burkholderia YNK-FB0053, the bacterial suspension and blank MST liquid culture medium were used as controls. The culture was cultured in a shaking incubator at 180 rpm at 30° C. for 7 days. The obtained culture solution sample was centrifuged at 12000 rpm for 10 min, and the supernatant was taken.

[0072] Turbidimetric method (absorbance) determination: Take 0.1g BaCl2, 1ml stabilizing solution, draw 250μL of the supernatant of the bacterial solution to be tested, add to a small beaker to make a 5ml system, stir for 1min on a magnetic stirrer, let stand for 15min. Use a UV-visible spectrophotometer to measure the absorbance at a wavelength of 420nm, repeat 3 times, adjust to zero by adding blank MST culture medium, and compare with the drawn standard curve.

[0073] The sulfate production rate was determined by spectrophotometer. The total amount of sulfate ions produced by the sulfur-oxidizing bacterial isolate was 66.538 ± 1.768 mg / L.

[0074] (II) Nitrogen fixation effect test

[0075] Preparation method of Asbhy nitrogen-free medium: weigh 10g glucose, 0.2g dipotassium hydrogen phosphate, 0.2g sodium chloride, 0.2g magnesium sulfate monohydrate, 0.2g potassium sulfate, 5g calcium carbonate, add to 1000mL water to dissolve, add 20g agar, and sterilize at 121℃ for 20min.

[0076] The strain YNK-FB0053 obtained in Example 1 was inoculated on the Aspergillus nitrogen-free culture plate by the four-point plate inoculation method, and the inoculation was repeated for 3 plates. After the inoculation, the plate was placed in a 30°C constant temperature incubator for 5 days, and the bacterial growth and the formation of transparent circles around the colonies were observed every day during the incubation period.

[0077] Figure 3 (B) shows the nitrogen fixation effect of strain YNK-FB0053. As can be seen from the figure, strain YNK-FB0053 can produce a transparent circle when cultured on Aspergillus nitrogen-free medium, indicating that the strain has the ability to fix nitrogen. The size of the transparent circle was measured by the cross method, and the ratio of the transparent circle diameter (D) to the colony diameter (d) was calculated to be D / d = 4.106 ± 0.695, indicating that strain YNK-FB0053 has a strong nitrogen fixation ability.

[0078] (III) Zinc dissolving effect test

[0079] Preparation method of solid culture medium: weigh 1g zinc oxide, 10g glucose, 0.2g potassium chloride, 1g ammonium sulfate, 0.2g magnesium sulfate, and 0.1g dipotassium hydrogen phosphate, add to 1000mL water to dissolve, adjust the pH to 7.3±0.1, add 20g agar, and sterilize at 121℃ for 20min.

[0080] The strain YNK-FB0053 obtained in Example 1 was inoculated on a zinc-soluble culture plate using the four-point plate inoculation method, and the inoculation was repeated for 3 plates. After the inoculation was completed, the plate was placed in a 30°C constant temperature incubator for 5 days, and the bacterial growth and the formation of transparent circles around the colonies were observed every day during the incubation period.

[0081] Figure 3 (C) shows the zinc dissolving effect of strain YNK-FB0053. As can be seen from the figure, strain YNK-FB0053 can produce a transparent circle when cultured on a zinc dissolving culture plate, indicating that the strain has the ability to dissolve zinc. The size of the transparent circle was measured using the cross method, and the ratio of the transparent circle diameter (D) to the colony diameter (d) was calculated to be D / d = 5.129 ± 0.459.

[0082] (IV) Phosphorus solubilization effect test

[0083] 1. Organophosphorus

[0084] Preparation method of organophosphorus solid culture medium: weigh 10g glucose, 0.5g ammonium sulfate, 0.5g yeast extract powder, 0.3g sodium chloride, 0.3g potassium chloride, 0.3g magnesium sulfate, 0.03g ferrous sulfate, 0.03g manganese sulfate, 0.2g lecithin, 1g calcium carbonate, add to 1000mL water to dissolve, adjust pH to 7.3±0.2, add 18±2g agar, and sterilize at 121℃ for 20min.

[0085] The strain YNK-FB0053 obtained in Example 1 was inoculated on an organophosphate culture plate using the four-point plate inoculation method, and the inoculation was repeated for three plates. After the inoculation was completed, the plate was placed in a 30°C constant temperature incubator for 5 days, and the bacterial growth and the formation of transparent circles around the colonies were observed every day during the incubation period.

[0086] Figure 3 (D) shows the effect of strain YNK-FB0053 on degrading organophosphate. As can be seen from the figure, strain YNK-FB0053 can produce a transparent zone when cultured on an organophosphate culture plate, indicating that the strain has the ability to degrade organophosphate. The size of the transparent zone was measured using the cross method, and the ratio of the transparent zone diameter (D) to the colony diameter (d) was calculated to be D / d = 3.453 ± 0.32.

[0087] 2. Inorganic phosphorus

[0088] Preparation method of inorganic phosphorus solid culture medium: weigh 10g glucose, 0.5g ammonium sulfate, 0.3g sodium chloride, 0.3g potassium chloride, 0.3g magnesium sulfate heptahydrate, 0.03g iron sulfate heptahydrate, 0.03g manganese sulfate tetrahydrate, 2g tricalcium phosphate, add to 1000mL water to dissolve, adjust pH to 7.3±0.1, add 20g agar, and sterilize at 121℃ for 20min.

[0089] The strain YNK-FB0053 obtained in Example 1 was inoculated on an inorganic phosphorus culture plate using the four-point plate inoculation method, and the inoculation was repeated for 3 plates. After the inoculation was completed, the plate was placed in a 30°C constant temperature incubator for 5 days, and the bacterial growth and the formation of transparent circles around the colonies were observed every day during the culture period.

[0090] Figure 3 (E) shows the effect of strain YNK-FB0053 on dissolving inorganic phosphorus. As can be seen from the figure, strain YNK-FB0053 can produce transparent circles when cultured on inorganic phosphorus culture plates, indicating that the strain has the ability to dissolve inorganic phosphorus. The size of the transparent circle was measured by the cross method, and the ratio of the transparent circle diameter (D) to the colony diameter (d) was calculated to be D / d = 2.06 ± 0.185.

[0091] (V) Siderophore production effect test

[0092] Iron-free Czapek medium (g / L): 30 g glucose, 3 g sodium nitrate, 1 g dipotassium hydrogen phosphate, 0.5 g potassium chloride, 0.5 g magnesium sulfate, 20 g agar, 1000 mL water.

[0093] CAS test solution: Solution A: 60.5 mg chrome azurol S and 10 ml Fe 3+ Solution (1mmol / LFeCl3·6H2O, 10mmol / LHCl) is added to 50ml water; Solution B: 72.9mg hexadecylamine bromide is added to 40ml water. Mix solution A and solution B after preparation.

[0094] Water agar: 18g agar, 1000ml water.

[0095] CAS lower layer color detection medium: After adding 10% CAS detection solution evenly to the 50-60℃ water agar medium, pour a larger amount of medium as the lower layer medium. After the water agar medium cools and solidifies, pour in the iron-free Czapek medium to form a double-layer plate.

[0096] The strain YNK-FB0053 obtained in Example 1 was inoculated into a CAS chromogenic double-layer culture medium using a 4-point inoculation method, and each treatment was repeated 3 times. The culture medium was placed in a 30°C incubator and the color development zone was observed continuously to see whether a color development zone was generated around the colony. The ratio between the diameter (D) of the color development zone generated and the colony diameter (d) was measured.

[0097] Figure 3 (F) shows the siderophore production effect of strain YNK-FB0053. As can be seen from the figure, the culture medium around the colony produces an orange-red halo, indicating that the strain can produce siderophores. The size of the color circle was measured by the cross method, and the ratio of the color circle diameter (D) to the colony diameter (d) was calculated to be D / d = 2.573 ± 0.265.

[0098] Example 3

[0099] This example is used to illustrate the relationship between Burkholderia gladiolus CCTCC NO: M20242288 and Fusarium solani ( Fusarium solani ), Fusarium equisetum ( Fusarium equiseti )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ) antagonistic effect.

[0100] Test pathogen: Fusarium solani ( Fusarium solani ), Fusarium equisetum ( Fusarium equiseti )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ) was provided by the laboratory of Institute of Agricultural Resources and Environment, Yunnan Academy of Agricultural Sciences.

[0101] Plate confrontation experiment: Fusarium solani ( Fusarium solani ), Fusarium equisetum ( Fusarium equiseti )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ) was used as the indicator pathogen. A 5 mm pathogen cake was inoculated in the center of the PDA medium, and the strain YNK-FB0053 obtained in Example 1 was inoculated in a cross shape at a distance of 25 mm from it. The uninoculated plate was used as a control. Each strain was replicated 3 times and cultured in a constant temperature incubator at 25-30°C in the dark for 5-7 days. The inhibition rate was calculated.

[0102] After the culture was completed, the average value and inhibition rate were calculated. Inhibition rate (%) = (colony diameter of the control group - colony diameter of the treatment group) / (colony diameter of the control group - 5) × 100.

[0103] Figure 5(A)-(D) show the activity of strain YNK-FB0053 against Fusarium solani ( Fusarium solani ), Fusarium equisetum ( Fusarium equiseti )、Fusarium rotundifolia( Fusarium concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ) antagonistic effect. The results of plate confrontation method showed that strain YNK-FB0053 could antagonize the above four pathogens, and the inhibition rate was above 50%. Fusarium concentricum ) had the best antagonistic effect, with an inhibition rate of 85.65%.

[0104] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. A strain of Burkholderia gladiolus ( Burkholderia gladioli ), characterized in that, The deposit number of this strain is CCTCC NO: M20242288.

2. A bacterial agent, characterized in that The active ingredient of the bacterial agent is the gladiolus Burkholderia described in claim 1.

3. Use of the gladiolus Burkholderia according to claim 1 or the bacterial agent according to claim 2 in sulfur oxidation and zinc dissolution, wherein sulfur oxidation refers to oxidation of sodium thiosulfate to produce sulfate ions; zinc dissolution refers to dissolving zinc oxide.

4. The gladiolus Burkholderia according to claim 1 or the bacterial agent according to claim 2 is effective in antagonizing Fusarium solani ( Fusarium solani ), Fusarium equisetum ( Fusarium equiseti )、Fusarium rotundifolia( Fusarium Concentricum ) and Brown Seashell Fungus ( Rosellinia necatrix ) in applications.

5. The bacterial agent according to claim 2, characterized in that: The bacterial agent is a liquid bacterial agent; the content of Burkholderia gladiolus in the bacterial agent is not less than 1×10 6 CFU / mL.

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