Bacillus preparation, bacillus compound preparation and application of bacillus preparation and bacillus compound preparation in increasing ferrophosphorus in soil

Abstract

The invention discloses a bacillus preparation, a bacillus compound preparation and application of the bacillus preparation and the bacillus compound preparation in increasing ferrophosphorus in soil, and belongs to the field of plant nutrition and microbiology. The preparation method comprises the following steps: centrifuging a bacterial solution of bacillus 1603IPR-02, and collecting supernate to obtain the siderophore preparation. The siderophore preparation is applied to crop root systems, southern soil or compounded with phosphate tailings to form a bacillus compound preparation to be applied to crops, plant growth can be promoted, plant biomass can be increased, meanwhile, the siderophore preparation has the efficient Fe-P activation capacity, Fe-P insoluble in soil is converted into phosphorus and iron capable of being utilized by plants, and the utilization rate of the Fe-P is increased. The bio-availability of soil nutrients is improved, and the absorption of crops on phosphorus and iron in soil is promoted. When the bacillus preparation or the bacillus compound preparation is used for treating pakchoi, the dry weight, the phosphorus content and the iron content of the pakchoi as well as the quick-acting phosphorus content and the effective iron content of soil can be increased. When applied to crops, the preparation has the characteristics of no pollution, no residue and biological environmental protection.

Description

Technical Field

[0001] The invention relates to the fields of plant nutrition and microbiology, and in particular to a bacillus preparation, a bacillus composite preparation and applications in improving phosphorus and iron in soil. Background Art

[0002] The problem of low phosphorus content is common in southern soils, and phosphorus is an indispensable element in the soil-plant-microorganism system. In the south, eluvial soils represented by yellow-brown soils and iron-rich soils represented by red soils are the most widely distributed, with a high degree of weathering, high iron and aluminum content, and strong phosphorus fixation. Studies have shown that in subtropical acidic soils, phosphorus combined with iron and aluminum oxides accounts for about 18% to 33% of the total phosphorus content. Although a large amount of phosphorus fertilizer is input into my country's farmland every year, the content that can be directly absorbed and utilized by plants is relatively low. In addition, the physical and chemical fixation and biological retention of phosphorus by soil and microbial systems have led to a serious accumulation of phosphorus in the soil. Inorganic phosphorus is the main form of phosphorus, and OP and Fe-P are the main inorganic phosphorus, which are difficult to be directly absorbed and utilized by plants. It is particularly important to improve the biological effectiveness of phosphorus and iron.

[0003] Phosphate tailings are byproducts produced during the phosphate mining and beneficiation process. Phosphate tailings contain SiO2, MgO, CaO, P2O5, Al2O3 and Fe2O3, but they are mainly composed of dolomite, fluoroapatite, etc., which are not easily directly absorbed and utilized by plants. Using phosphate tailings as fertilizers for agricultural production is a way to solve the problem of phosphate tailings, but how to efficiently activate the nutrients in phosphate tailings resources is a major problem that needs to be solved urgently. There are many related studies on the activation of phosphate tailings, such as physical and mechanical activation and chemical acid activation. However, there are currently few studies on the use of biological advantages to activate the nutrients in phosphate tailings using microorganisms and their secretions.

[0004] Microorganisms will secrete microbial siderophores under iron deficiency conditions. They are a class of small molecules that have a high affinity for trace elements and can be absorbed and utilized by microbial cells after chelating with insoluble trace elements in the environment. It is generally believed that microorganisms dissolve inorganic phosphorus by secreting low molecular weight organic acids, such as lactic acid, oxalic acid, malic acid, citric acid, etc. The mechanism of organic acid activation of phosphorus is as follows: 1. Acid dissolution; 2. Release of phosphate ions by chelating Fe, Al, Ca, etc.; 3. Reduce the adsorption of phosphate by soil by competing for adsorption sites on the soil surface; 4. Increase the activity of soil enzymes such as acid phosphatase to increase the effectiveness of phosphorus. After the siderophores secreted by microorganisms chelate with iron in Fe-P, they can release insoluble phosphorus, thereby improving the effectiveness of phosphorus and iron in the soil. Relevant studies have shown that the application of iron-reducing bacteria can increase the content of free iron oxide in the soil and increase the content of Resin-P (resin exchangeable phosphorus) in the soil, thereby activating iron and phosphorus in the soil.

[0005] Most of the reports on the activation of Fe-P in southern soils use small molecule organic acids, and no relevant research has been conducted using microbial iron carriers. The present invention uses Bacillus and the iron carriers it produces for the activation of phosphorus in southern soils for the first time. Compared with the previously conducted culture medium phosphorus dissolution test (CN115109710A, CN113717885A), the existence state of its activated phosphorus element is completely different. The present invention uses microorganisms and their secretion iron carriers to efficiently activate effective phosphorus and iron in southern soils. Not only does the microorganism itself have a phosphorus-dissolving effect, but the secretion product iron carriers cultivated using iron-deficient culture medium also have this effect. In agriculture, the use of microorganisms and their secreted iron carriers to chelate iron and release phosphorus can reduce the application of chemical fertilizers, improve the effectiveness of iron and phosphorus in southern soils, is an energy-saving, green, and healthy fertility supplement method, has important ecological and economic significance for the development of agriculture, and provides a basis for the future development of bacterial fertilizers with improved plant phosphorus nutrition and trace element functions. Summary of the invention

[0006] The purpose of the present invention is to provide a bacillus preparation, a bacillus composite preparation and use thereof in improving phosphorus and iron in soil, so as to solve the problems existing in the above-mentioned prior art. The bacillus preparation or the bacillus composite preparation prepared by the present invention solves the problem that phosphorus and iron in the soil are difficult to be absorbed by crops due to the large amount of input of phosphorus fertilizer but the low absorption rate of crops.

[0007] To achieve the above object, the present invention provides the following solutions:

[0008] The present invention provides use of a bacillus preparation in any of the following:

[0009] (1) Application in increasing available phosphorus and / or effective iron in southern soils;

[0010] (2) Application in increasing the dry weight of Chinese cabbage;

[0011] (3) Application in increasing the phosphorus content of Chinese cabbage;

[0012] (4) Application in the preparation of biological fertilizers.

[0013] The bacillus preparation is composed of Bacillus sp. 1603IPR-02 or its bacterial solution or its secreted iron carrier as an effective ingredient, and the deposit number of the Bacillus sp. 1603IPR-02 is CGMCC No.21640.

[0014] The above-mentioned iron carrier is a hydroxamic acid-containing iron carrier.

[0015] Preferably, the iron carrier secreted by Bacillus 1603IPR-02 is obtained by centrifuging the fermentation broth of Bacillus 1603IPR-02 and collecting the supernatant to obtain the Bacillus 1603IPR-02 iron carrier.

[0016] Preferably, the step of obtaining the fermentation broth comprises: inoculating Bacillus 1603IPR-02 into SSM iron-deficient culture medium for constant temperature shaking culture.

[0017] Preferably, the Bacillus is applied to the roots of Chinese cabbage by root application, and the amount of the Bacillus preparation used for each Chinese cabbage is 50 mL.

[0018] The present invention also provides the use of the Bacillus composite preparation in any of the following items:

[0019] (1) Application in increasing available phosphorus and / or effective iron in southern soils;

[0020] (2) Application in increasing the dry weight of Chinese cabbage;

[0021] (3) Application in increasing the phosphorus content of Chinese cabbage;

[0022] (4) Application in the preparation of biological fertilizers;

[0023] The Bacillus composite preparation is composed of (a) any one of Bacillus 1603IPR-02, Bacillus 1603IPR-02 bacterial solution and Bacillus 1603IPR-02 iron carrier; and (b) phosphate tailings as active ingredients, and the preservation number of Bacillus 1603IPR-02 is CGMCC No.21640.

[0024] Preferably, the Bacillus 1603IPR-02 iron carrier is obtained by inoculating Bacillus 1603IPR-02 into SSM iron-deficient medium for fermentation, centrifuging the fermentation broth, and collecting the supernatant.

[0025] Preferably, the Bacillus composite preparation uses Bacillus 1603IPR-02 iron carrier and phosphate tailings as active ingredients, and the ratio of the Bacillus 1603IPR-02 iron carrier and phosphate tailings is 1mL:5g; or uses Bacillus 1603IPR-02 bacterial solution and phosphate tailings as active ingredients, and the ratio of the Bacillus 1603IPR-02 bacterial solution and phosphate tailings is 1mL:5g.

[0026] Preferably, the method of using the Bacillus composite preparation includes the following steps: applying the phosphate tailings in the Bacillus composite preparation in advance into the soil for planting Chinese cabbage at a mass ratio of 1:1, and then after thinning the Chinese cabbage, applying 50 mL of Bacillus 1603IPR-02 iron carrier to the roots of each Chinese cabbage plant once every 7 days.

[0027] The present invention also provides a biological fertilizer, whose effective ingredients are (1) the bacterial liquid of the Bacillus 1603IPR-02 and phosphate tailings, the ratio of the bacterial liquid of the Bacillus 1603IPR-02 and the phosphate tailings is 1mL:5g; or (2) the iron carrier secreted by the Bacillus 1603IPR-02 and the phosphate tailings, the ratio of the iron carrier and the phosphate tailings is 1mL:5g.

[0028] Preferably, the biological fertilizer is used to increase the available phosphorus and / or effective iron in southern soil, or to increase the dry weight and / or phosphorus content of Chinese cabbage.

[0029] The present invention discloses the following technical effects:

[0030] The bacillus 1603IPR-02 disclosed in the present invention is isolated from plant rhizosphere soil. The present invention applies the bacterial liquid of the strain or the iron carrier secreted by the strain to the soil in the south or the planting of Chinese cabbage. It is found that the strain has the ability to efficiently activate Fe-P, and can convert the soil insoluble Fe-P into phosphorus and iron that can be used by plants, increase the plant effectiveness of soil nutrients, and the experimental results show that the bacillus and its secreted products can promote the absorption of soil phosphorus by Chinese cabbage, improve plant phosphorus and iron nutrition, and increase the yield and phosphorus content of Chinese cabbage. The present invention also applies the bacteria and phosphorus tailings in combination to the soil in the south and the planting of Chinese cabbage. The experimental results show that the soil insoluble Fe-P can also be converted into phosphorus and iron that can be used by plants, increase the plant effectiveness of soil nutrients, and increase the yield and phosphorus content of Chinese cabbage. The preparation prepared by using Bacillus 1603IPR-02 or the preparation prepared by combining it with phosphate tailings in the present invention is pollution-free and residue-free during the application process, and is biologically environmentally friendly, indicating that Bacillus 1603IPR-02 is a growth-promoting strain with good application prospects in the field of plant growth promotion, and can also efficiently activate the resource components of phosphate tailings. In addition, the present invention also improves the preparation method of the iron carrier, simplifies the production steps, reduces the production cost, and makes it more convenient to obtain the iron carrier, which is beneficial to the promotion of industrialization. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0032] Figure 1 This is the effect of Bacillus 1603IPR-02 and its siderophore on the dry weight of Chinese cabbage in Example 1 of the present invention;

[0033] Figure 2 This is the effect of Bacillus 1603IPR-02 and its siderophore on the phosphorus content of Chinese cabbage in Example 1 of the present invention;

[0034] Figure 3 This is the effect of Bacillus 1603IPR-02 and its siderophore on the available phosphorus content in soil in Example 1 of the present invention;

[0035] Figure 4 This is the effect of Bacillus 1603IPR-02 and its iron carrier on the effective iron content in soil in Example 1 of the present invention;

[0036] Figure 5 This is the effect of Bacillus 1603IPR-02 and its iron carrier compounded with phosphorus tailings on the dry weight of Chinese cabbage in Example 2 of the present invention;

[0037] Figure 6 This is the effect of Bacillus 1603IPR-02 and its iron carrier compounded with phosphorus tailings on the phosphorus content of Chinese cabbage in Example 2 of the present invention;

[0038] Figure 7 This is the effect of Bacillus 1603IPR-02 and its iron carrier compounded with phosphate tailings on the available phosphorus content in soil in Example 2 of the present invention;

[0039] Figure 8 This is the effect of Bacillus 1603IPR-02 and its iron carrier compounded with phosphate tailings on the effective iron content in soil in Example 2 of the present invention;

[0040] In the above figure, clear water represents the blank control, Bac solution represents the experimental treatment group in which Bacillus 1603IPR-02 solution is applied to the root system of Chinese cabbage or applied to the soil in the south, Bac siderophore represents the experimental treatment group in which Bacillus 1603IPR-02 siderophore is applied to the root system of Chinese cabbage or applied to the soil in the south, Bac solution + phosphate tailings represents the experimental treatment group in which Bacillus 1603IPR-02 solution and phosphate tailings are compounded and applied to the root system of Chinese cabbage or applied to the soil in the south, Bac siderophore + phosphate tailings represents the experimental treatment group in which Bacillus 1603IPR-02 siderophore and phosphate tailings are compounded and applied to the root system of Chinese cabbage or applied to the soil in the south. DETAILED DESCRIPTION

[0041] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0042] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0043] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0044] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0045] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0046] Unless otherwise specified, the culture medium used in the following examples is formulated as follows:

[0047] (1) SSM medium

[0048] Dissolve 1g (NH4)2SO4, 4g C4H6O4 and 0.2g MgSO4 in 1000mL deionized water. Dissolve 6g K2HPO4 and 3g KH2PO4 in 20mL deionized water, adjust pH to 7.0 with potassium hydroxide solution, sterilize at 121℃ for 20min, then mix and add.

[0049] (2)CAS test solution

[0050] CAS detection solution: prepare 2mM CAS stock solution and 1mM FeCl3 stock solution respectively. Weigh 0.0219g CTAB and dissolve it in 25mL ultrapure water for later use. Weigh 4.3079g anhydrous piperazine and dissolve it in 30mL ultrapure water, and adjust pH to 5.6 with 12M concentrated hydrochloric acid to obtain piperazine buffer. Mix 7.5mL of 2mM CAS solution and 1.5mL of 1mM FeCl3, and slowly add it to 25mL CTAB solution while stirring. Stir slowly to prevent excessive foaming, and then add 30mL piperazine buffer to mix. Before using the detection solution, weigh 0.0873g 5-sulfosalicylic acid and add it, transfer it to a 100mL volumetric flask, make up to volume and gently shake to mix.

[0051] The Bacillus sp. 1603IPR-02 used in the following examples has a deposit number of CGMCC No. 21640. The bacterium has also been disclosed in the invention patent CN115109710B, Bacillus sp. 1603IPR-02 with the ability to produce siderophores and its application.

[0052] Example 1 Effects of Bacillus 603IPR-02 and iron carriers on the growth of Chinese cabbage

[0053] 1. Test materials

[0054] Bacillus 1603IPR-02 bacterial solution, crop (Brachys pakchoi), soil (southern red soil), base fertilizer (NPK: 100-50-50mg / kg).

[0055] 2. Test methods

[0056] The strain in the logarithmic growth phase was inoculated into SSM iron-deficient medium, and cultured at 30°C for 48 hours to obtain the fermentation liquid, which was centrifuged in a high-speed refrigerated centrifuge at 4°C and 10,000r for 5 minutes to obtain the iron carrier supernatant. The iron carrier yield in the supernatant was detected using CAS detection solution, and the iron carrier yield was 150-200 μmol / L.

[0057] A pot experiment was set up with 3 treatments: ① CK water treatment; ② application of strain fermentation liquid (50 mL / plant); ③ application of iron carrier supernatant (50 mL / plant). Root application, 4 biological replicates for each treatment. Application was carried out three days after thinning, once every 7 days, and 4 times in total during the entire growth period. The pakchoi was harvested at 40 days of growth, and relevant data were measured.

[0058] 3. Test results

[0059] The results showed that the application of Bacillus 1603IPR-02 and its iron carrier significantly increased the dry weight of Chinese cabbage (see Figure 1 ), with increases of 15.7% and 24.6% respectively; it significantly promoted the plant's absorption of phosphorus, with increases of 18.5% and 9.3% respectively (see Figure 2 ), which has a significant promoting effect on improving plant yield and nutrition.

[0060] Application of Bacillus 1603IPR-02 and its siderophore also significantly increased soil available phosphorus and available iron content, and soil available phosphorus increased by about 3 times and 2 times, respectively (see Figure 3 ), the effective iron content in the soil increased by 22.8% and 18.6% (see Figure 4 ), indicating that it has a very good chelating activation effect on Fe-P.

[0061] Example 2 Effect of Bacillus strain 1603IPR-02 and iron carrier combined with phosphate tailings on the growth of Chinese cabbage

[0062] 1. Test materials

[0063] Bacillus 1603IPR-02 bacterial solution, crop (Brachys pakchoi), soil (southern red soil), phosphorus tailings, base fertilizer (NPK: 100-50-50mg / kg).

[0064] 2. Test methods

[0065] A pot experiment was set up with 4 treatments: ①CK water treatment; ②phosphate tailings treatment; ③Application of strain fermentation liquid + phosphate tailings (the ratio of strain fermentation liquid (mL) to phosphate tailings (g) was 1:5); ④Application of iron carrier supernatant + phosphate tailings (the ratio of iron carrier supernatant (mL) to phosphate tailings (g) was 1:5); root application. For the treatment involving phosphate tailings, the phosphate tailings were mixed with the soil in a 1:1 ratio before arranging the potted plants, and no phosphate tailings were added subsequently. Each treatment had 4 biological replicates. Application was carried out three days after thinning, once every 7 days, and a total of 4 times during the entire growth period. The pakchoi was harvested after 40 days of growth, and the relevant data were measured.

[0066] 3. Test results

[0067] Applying phosphate tailings as fertilizer alone can also significantly promote the growth of Chinese cabbage and significantly increase the phosphorus content of the plants. After mixing, the content of available phosphorus and effective iron in the matrix is ​​significantly higher than that in normal soil. On the basis of adding phosphate tailings, adding Bacillus 1603IPR-02 or iron carrier activates the phosphate tailings and soil insoluble nutrients, further promoting the growth and development of Chinese cabbage. Plant dry weight: Compared with the CK water treatment, treatments ③ and ④ increased by 36.5% and 38.9%, respectively. Compared with the phosphate tailings treatment, treatments ③ and ④ increased by 12.3% and 14.2%, respectively (see Figure 5 ); Plant phosphorus content: Compared with CK water treatment, ③④ increased by about 52% and 66% respectively, compared with phosphorus tailings treatment, ③④ increased by 14.6% and 25.1% respectively (see Figure 6 ); Soil available phosphorus: Compared with CK water treatment, the two treatments increased by about 4 times, and compared with phosphorus tailings treatment, it increased by about 2 times (see Figure 7 ); Soil available iron: Compared with CK water treatment, ③ and ④ increased by 74.6% and 61.4%, respectively, and compared with phosphate tailings treatment, increased by 11.5% and 5.3%, respectively (see Figure 8 ).

[0068] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Use of the Bacillus preparation in any of the following: (1) Application in increasing available phosphorus and / or effective iron in southern soils; (2) Application in increasing the dry weight of Chinese cabbage; (3) Application in increasing the phosphorus content of Chinese cabbage; (4) Application in the preparation of biological fertilizers. in, The bacillus preparation uses bacillus sp. 1603IPR-02 or its bacterial solution or iron carrier secreted by it as an effective ingredient. The deposit number of the bacillus sp. 1603IPR-02 is CGMCC No.21640.

2. The use according to claim 1, characterized in that The iron carrier secreted by Bacillus 1603IPR-02 is obtained by centrifuging the fermentation liquid of Bacillus 1603IPR-02 and collecting the supernatant to obtain the iron carrier of Bacillus 1603IPR-02.

3. The use according to claim 2, characterized in that The step of obtaining the fermentation broth comprises: inoculating Bacillus 1603IPR-02 into SSM iron-deficient culture medium for constant temperature shaking culture.

4. The use according to claim 1, characterized in that The bacillus is applied to the roots of Chinese cabbage in a root application manner, and the amount of the bacillus preparation used for each Chinese cabbage is 50 mL.

5. Use of the Bacillus composite preparation in any of the following: (1) Application in increasing available phosphorus and / or effective iron in southern soils; (2) Application in increasing the dry weight of Chinese cabbage; (3) Application in increasing the phosphorus content of Chinese cabbage; (4) Application in the preparation of biological fertilizers; in, The bacillus composite preparation is composed of (a) any one of bacillus 1603IPR-02, bacillus 1603IPR-02 bacterial solution and bacillus 1603IPR-02 iron carrier; and (b) phosphate tailings as effective ingredients, and the preservation number of the bacillus 1603IPR-02 is CGMCC No.21640.

6. The use according to claim 5, characterized in that The Bacillus 1603IPR-02 iron carrier is obtained by inoculating Bacillus 1603IPR-02 into SSM iron-deficient culture medium for fermentation, centrifuging the fermentation liquid, and collecting the supernatant.

7. The use according to claim 5, characterized in that The Bacillus composite preparation uses Bacillus 1603IPR-02 iron carrier and phosphate tailings as active ingredients, and the ratio of the Bacillus 1603IPR-02 iron carrier and phosphate tailings is 1mL:5g; or uses Bacillus 1603IPR-02 bacterial solution and phosphate tailings as active ingredients, and the ratio of the Bacillus 1603IPR-02 bacterial solution and phosphate tailings is 1mL:5g.

8. The use according to claim 7, characterized in that The method of using the Bacillus composite preparation comprises the following steps: applying the phosphate tailings in the Bacillus composite preparation into the soil for planting Chinese cabbage in advance according to a mass ratio of 1:1, and then applying 50 mL of Bacillus 1603IPR-02 iron carrier to the roots of each Chinese cabbage plant after thinning the Chinese cabbage seedlings, once every 7 days.

9. A biological fertilizer, characterized in that: The active ingredients are (1) the bacterial solution of Bacillus 1603IPR-02 and phosphate tailings as described in claim 1, wherein the ratio of the bacterial solution of Bacillus 1603IPR-02 and the phosphate tailings is 1 mL:5 g; or (2) the iron carrier secreted by Bacillus 1603IPR-02 as described in claim 1 and phosphate tailings, wherein the ratio of the iron carrier and the phosphate tailings is 1 mL:5 g.

10. The biological fertilizer according to claim 9, characterized in that The biological fertilizer is used for increasing the available phosphorus and / or effective iron in southern soil, or increasing the dry weight and / or phosphorus content of Chinese cabbage.

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