CN111849842B - Potassium bacteria, potassium bacteria microbial inoculum comprising same and application - Google Patents

Potassium bacteria, potassium bacteria microbial inoculum comprising same and application Download PDF

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CN111849842B
CN111849842B CN202010828032.6A CN202010828032A CN111849842B CN 111849842 B CN111849842 B CN 111849842B CN 202010828032 A CN202010828032 A CN 202010828032A CN 111849842 B CN111849842 B CN 111849842B
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potassium bacteria
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罗雯
万全玉
张同林
刘旺喜
饶友生
石路怀
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Abstract

The invention discloses a potassium bacteria, a potassium bacteria microbial inoculum comprising the same and application thereof. The taxonomy of the potassium-solubilizing bacteria is named as Burkholderia K4-1, and the preservation number is CCTCC M2020187. The potassium-solubilizing bacterial strain is obtained by independent separation of the inventor, is easy to grow in a used culture medium, can be massively propagated, has a growth promoting effect on plants, and simultaneously has an inhibiting effect on the growth of bacteria and moulds in soil.

Description

Potassium bacteria, potassium bacteria microbial inoculum comprising same and application
Technical Field
The invention relates to the field of microorganisms, and particularly relates to a potassium bacteria, a potassium bacteria microbial inoculum comprising the same and application thereof.
Background
Potassium is one of three elements of crop nutrition, is commonly present in crops, has functions related to plant metabolism, widely influences the growth and metabolism of crops in the form of enzyme activators, can activate enzymes, and currently, about 60 enzymes are found to be related to potassium, and have very important roles in physiological processes such as light energy utilization, sugar metabolism, protein synthesis, cell osmotic regulation, plant resistance enhancement and the like.
The soil in China is rich in potassium, but the potassium exists in the forms of mineral potassium and fixed potassium and cannot be directly absorbed by plant roots. The soil is seriously lack of effective potassium, and the soil becomes one of important factors for limiting agricultural production in China.
The potassium-dissolving bacteria are bacteria which are separated from soil and can differentiate aluminosilicate and apatite minerals, can be used as microbial fertilizers, can decompose insoluble aluminosilicate inorganic minerals such as potassium feldspar and apatite, promote nutrient elements such as insoluble potassium, phosphorus, silicon and magnesium to be converted into soluble nutrients, increase the content of quick-acting nutrients in soil, promote crop growth and development and improve yield. However, at present, the potassium-solubilizing bacterial manure mainly uses bacillus mucilaginosus as a main functional strain, and researches on potassium-solubilizing functions of other strains are shallow and are only reported.
Disclosure of Invention
The invention aims to provide a potassium bacteria, a potassium bacteria microbial inoculum comprising the same and application thereof, so as to provide a novel potassium bacteria microbial inoculum.
In order to achieve the above object, according to one aspect of the present invention, there is provided a potassium solubilizing bacterium. The taxonomic name of the potassium-solubilizing bacterium is Burkholderia K4-1, and the preservation number is CCTCC M2020187.
According to another aspect of the present invention, there is provided a potassium bacteria inoculant for soil. The potassium bacteria inoculant for soil comprises the potassium bacteria.
Further, the potassium bacteria inoculant is prepared by the following steps: activating potassium-decomposing bacteria, inoculating the potassium-decomposing bacteria on an activation culture medium, putting the activated culture medium into a constant-temperature incubator, and performing constant-temperature culture for 3-4 days at the temperature of 28 ℃; selecting strains on the activated culture medium, expanding and inoculating the strains on a potassium-solubilizing culture medium, culturing for 3-4 days at a constant temperature of 28 ℃, and washing the lawn to prepare a bacterial suspension, namely the potassium-solubilizing bacterium agent.
Further, the potassium bacteria inoculant is prepared by the following steps: activating potassium-decomposing bacteria, inoculating the potassium-decomposing bacteria on an activation culture medium, putting the activated culture medium into a constant-temperature incubator, and performing constant-temperature culture for 3-4 days at the temperature of 28 ℃; and (3) selecting strains on the activation culture medium, inoculating the strains in a fermentation culture medium, performing shaking culture at 28 ℃ for 6-7 days to obtain a fermentation liquid, centrifuging, collecting precipitates, and performing water resuspension to obtain the potassium bacteria killing agent.
Further, the potassium bacteria slant culture medium is preserved.
Further, the step of centrifugally collecting the precipitate comprises the step of centrifuging at 5000-6000 r/min for 8-10 min and collecting the precipitate.
Further, the activation medium is an alexander bauov solid medium.
Further, the fermentation medium comprises the following components in parts by weight: the fermentation medium comprises the following components in parts by weight: 5-10 g of cane sugar and (NH)4)2SO4 0.5~1g、Na2HPO4·12H2O 7.5~12.5g、MgSO4·7H2O 0.75~1.25g、CaCO30.5-1 g, 1-2 g potassium feldspar powder and 1000mL distilled water, preferably 10g sucrose, (NH)4)2SO4 1g、Na2HPO4·12H2O 10g、MgSO4·7H2O 0.75g、CaCO30.5g, 2g of potassium feldspar powder and 1000mL of distilled water, adjusting the pH value of the culture medium to 7.5-7.9, and sterilizing at 121 ℃ for 20min for later use.
Further, washing the potassium feldspar powder with deionized water for 3-5 times, and naturally drying in the shade.
According to another aspect of the invention, the application of the potassium bacteria or the potassium bacteria microbial inoculum for soil in soil potassium removal is provided.
The potassium-solubilizing bacterial strain is obtained by independent separation of the inventor, is easy to grow in a used culture medium, can be massively propagated, has a growth promoting effect on plants, and simultaneously has an inhibiting effect on the growth of bacteria and moulds in soil.
Information on strain preservation
The strains used by the invention are all preserved in China Center for Type Culture Collection (CCTCC) with the preservation addresses: china, wuhan university, zip code: 430072; telephone: (027) -68754052. The specific strain preservation information is as follows:
burkholderia K4-1(Burkholderia cenocepacia K4-1) was deposited in China Center for Type Culture Collection (CCTCC) at 6, 8 and 2020 with the deposit number of CCTCC NO: M2020187.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this application, illustrate embodiments of the invention and, together with the description, serve to explain the invention and not to limit the invention. In the drawings:
FIG. 1 shows the potassium-solubilizing circle of the potassium-solubilizing bacterium in example 1;
FIG. 2 shows Burkholderia K4-116S rDNA phylogenetic tree;
FIG. 3 shows a scanning electron micrograph of unfermented potassium feldspar powder in example 1; and
FIG. 4 shows an electron micrograph of potassium feldspar powder after fermentation in example 1.
Detailed Description
It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail below with reference to the embodiments with reference to the attached drawings.
The inventor of the invention screens a new potassium-decomposing bacterium from a soil sample of red soil in the west of Jiangxi. The taxonomic name of the potassium-solubilizing bacterium is Burkholderia K4-1, and the preservation number is CCTCC M2020187. The potassium-solubilizing bacterial strain is obtained by independent separation of the inventor, is easy to grow in a used culture medium, can be massively propagated, has a growth promoting effect on plants, and simultaneously has an inhibiting effect on the growth of bacteria and moulds in soil.
In order to make the potassium bacteria fully utilized, according to an exemplary embodiment of the present invention, a potassium bacteria inoculant for soil is provided. The potassium bacteria inoculant comprises the potassium bacteria. In a typical embodiment of the invention, the potassium bacteria inoculant is prepared by the following steps: activating potassium-decomposing bacteria, inoculating the activated potassium-decomposing bacteria on an activation culture medium, putting the activated potassium-decomposing bacteria into a constant-temperature incubator, and performing constant-temperature culture at 28 ℃ for 3-4 days; selecting strains on the activated culture medium, expanding and inoculating the strains on a potassium-solubilizing culture medium, culturing for 3-4 days at a constant temperature of 28 ℃, and washing the lawn to prepare a bacterial suspension, namely the potassium-solubilizing bacterium agent. In another exemplary embodiment of the present invention, the potassium bacteria inoculant is prepared by the following steps: activating potassium-decomposing bacteria, inoculating the potassium-decomposing bacteria on an activation culture medium, putting the activated culture medium into a constant-temperature incubator, and performing constant-temperature culture for 3-4 days at the temperature of 28 ℃; selecting strains on an activation culture medium, inoculating the strains in a fermentation culture medium, carrying out shake culture at 28 ℃ for 6-7 days to obtain a fermentation liquid, centrifuging, collecting precipitates, and carrying out water resuspension to obtain the potassium bacteria decomposing agent.
Preferably, the potassium bacteria slant culture medium is preserved, and preferably, the centrifugation and collection of the sediment comprises centrifugation at 6000r/min for 10min and collection of the sediment.
According to a typical embodiment of the invention, the activation medium is an alexander bauov solid medium, comprising in parts by weight: sucrose 5g, Na2HPO4·12H2O 2g、MgSO4·7H2O 0.5g、FeCl3 0.005g、CaCO30.1g, 1g of potassium feldspar powder (natural dried sample after 5 times of deionized water cleaning), and 1000mL of distilled water; adjusting the pH of the culture medium to 7.0-7.5, and sterilizing at 121 deg.C for 20 min. The fermentation medium comprises the following components in parts by weight: the fermentation medium comprises the following components in parts by weight: 5-10 g of cane sugar and (NH)4)2SO4 0.5~1g、Na2HPO4·12H2O 7.5~12.5g、MgSO4·7H2O 0.75~1.25g、CaCO30.5-1 g of potassium feldspar powder, 1-2 g of potassium feldspar powder and 1000mL of distilled water; preferably sucrose 10g, (NH)4)2SO4 1g、Na2HPO4·12H2O 10g、MgSO4·7H2O 0.75g、CaCO30.5g, 2g of potassium feldspar powder and 1000mL of distilled water, adjusting the pH value of the culture medium to 7.5-7.9, and sterilizing at 121 ℃ for 20min for later use. The potassium-dissolving culture medium (slant storage culture medium) is prepared by adding agar 20g to the above fermentation culture medium.
According to a typical embodiment of the invention, the application of the potassium bacteria or the potassium bacteria microbial inoculum for soil in soil potassium removal is also provided.
The following will further illustrate the beneficial effects of the present invention by combining experimental data and examples.
Example 1
1. Separating soil potassium bacteria: spreading the suspension on Alexandrium bauhinia solid culture medium (sucrose 0.5g, Na)2HPO4 0.2g、MgSO4-7H2O 0.05g、FeCl3 0.0005g、CaCO30.01g, potassium feldspar powder 0.1g, agar 2.0g, distilled water 100mL, pH 7.0-7.5, sterilizing at 121 ℃ for 20min), air drying, and inversely culturing in a constant temperature incubator at 28 ℃ for 3-4 d. Selecting a viscous and drop-shaped bacterial colony in a plate culture medium, and purifying to obtain a single bacterial colony. Taking a small amount of thallus and dibblingCulturing in a 28 deg.C incubator for 4d on a double-layer medium (the upper layer is primary-screening medium and the lower layer is 2% agar) to obtain potassium-dissolving ring, as shown in FIG. 1.
The bacterial colony of the strain on an Alexander bauov solid culture medium is milk white, smooth and full, neat in edge, sticky and capable of being pulled into a filament. The thalli is gram-negative, short rod-shaped, without capsule and spore and with motility through microscopic examination.
Through 16S rDNA sequence analysis, sequencing results are compared with standard sequences in a GeneBank database through BLAST, a strain 16S rDNA sequence with high similarity is selected, then MEGA6.0 is used for constructing a strain phylogenetic tree, the strain is closest to the genetic relationship with Burkholderia, the similarity is 99%, and the strain is named as Burkholderia K4-1. Burkholderia K4-116S rDNA phylogenetic tree is shown in FIG. 2.
2. Analysis of potassium-resolving activity of Burkholderia K4-1: 500. mu.L of Burkholderia K4-1 bacterial suspension (bacterial concentration 10)8) Inoculated into a 100mL liquid medium (sucrose 1g, (NH)4)2SO4 0.1g、Na2HPO4·12H2O 1g、MgSO4·7H2O 0.075g、CaCO30.05g, potassium feldspar powder 0.2g, distilled water 100mL, pH 7.9, sterilization at 121 ℃ for 20min) in a 250mL conical flask, three parallel flasks were made, and no inoculation control was set. Shaking at 28 deg.C and 200r/min for 6-7d, and the viable count in the culture can reach 109cfu/mL. Adding 6% H by volume fraction into the fermentation liquor2O2Digesting at 120 deg.C, and ultrasonically crushing for 2min with cells after viscous substances in the fermentation solution completely disappear, 20/40 s. The lysate is centrifuged in a high speed centrifuge at 12000r/min for 10min, the precipitate is separated from the supernatant and the supernatant is filtered through a 0.45mm filter membrane. By flame spectrophotometer (determination of potassium and sodium in water quality, flame atomic absorption photometry GB 11904-89)]The content of effective potassium in the filtrate was measured, and the content of effective potassium in the fermentation broth was increased by 29.8% (with blank medium as control, sucrose 1.0g, (NH)4)2SO40.1g、Na2HPO4·12H2O 1g、MgSO4-7H2O 0.075g、CaCO30.05g, potassium feldspar powder 0.2g, distilled water 100mL, pH 7.9, 121 ℃ sterilization for 20 min).
Taking the fermentation liquor of Burkholderia K4-1 cultured for 10 days, collecting potassium feldspar powder, and analyzing by a scanning electron microscope, wherein the potassium feldspar powder crystallization shows that the obvious corrosion change is shown in figure 4. The potassium feldspar powder without fermentation treatment is shown in figure 3.
3. Effect of burkholderia K4-1 on plant growth in outdoor field trials: test decomposition of Potassium bacterium group (the number of applied bacteria per square meter of field is about 10)10And (3) base fertilizer: 30g of urea and 9.375g of diammonium hydrogen phosphate/m2) And CK group (blank control group, no inoculation of any strain, base fertilizer: 30g of urea and 9.375g of diammonium hydrogen phosphate/m2). Each group comprises 1 experimental field, each experimental field is 1m2And sowing 36 germinated vegetable seeds. Each experimental field was periodically watered with an equal amount of deionized water. The vegetable variety is Chinese little greens (seed producer: seed breeding station of Wangzhengshen shop in Qingxian county) or Chinese cabbage (seed producer: vegetable seedling research institute in Shandong Lingzi No. 26384m), and is harvested 40 days after sowing. Detecting the chlorophyll content of leaves by spectrophotometry, detecting the reducing sugar content of plants by anthrone colorimetry, and detecting the protein content of leaves by Coomassie Brilliant blue G-250 method. The results after statistical analysis show that the chlorophyll content of the Shanghai chicken feather meal is increased by 42.58% in the vegetables obtained by the experimental field cultivation applying the bacterial suspension compared with the detection result of a blank control group; the protein content in Chinese cabbage is increased by 159.33%. The nutritive value of the vegetables is obviously improved. After 2 months, the content of the available potassium in the soil is increased (the increase is 9.6%).
4. Preparing beef extract peptone solid agar plate, potato glucose agar plate, and Gao's I solid agar plate (adding 10% phenol solution to inhibit growth of bacteria and mold); and (3) diluting the soil of the potassium bacteria decomposing group and the CK group to prepare suspensions, coating the suspensions on three solid agar medium plates, culturing at a constant temperature of 28 ℃ until colonies grow out, and recording the number of bacteria, actinomycetes and moulds. The statistical analysis result shows that the number of the mould and the bacteria in the soil of the potassium bacteria decomposing group is reduced, and the number of the actinomycetes is relatively increased compared with the CK group.
TABLE 1 Effect of Potassium solubilizing Agents on soil microbial communities
Figure GDA0002688421730000041
Figure GDA0002688421730000051
Example 2
Soil for test is prepared in a nursery pot, and potassium bacteria group for test is decomposed (0.28 g of urea, 0.875g of diammonium hydrogen phosphate and Burkholderia K4-1 bacterial suspension 10 are applied to each kilogram of soil)8) And CK group (blank control group, no inoculation of any strain, base fertilizer 0.28g urea and diammonium hydrogen phosphate 0.875g per kilogram of soil). Repeating 5 pots in each group, storing at room temperature for 1 year, adding the K4-1 bacterial suspension 10 into the soil of potassium-decomposing bacteria group85 germinated vegetable seeds are sown, and after the seeds germinate, thinning the seeds to 3. Periodically, each pot of soil is watered with the same amount of deionized water. The vegetable variety is Shanghai herba Elsholtziae Penduliforare (seed producer: seed breeding station of Wangzhen town of Qingcounty), and is harvested 30 days after sowing. The plants were cut along the roots, washed with water and the biomass was measured. As can be seen from Table 2, the potassium-solubilizing bactericide can obviously improve the plant height and fresh weight of the Shanghai Chinese little greens, and increase the yield by 34.27% compared with the case of not adding the bactericide, which indicates that the potassium-solubilizing bactericide can obviously improve the growth characteristics and yield of the Shanghai Chinese little greens. The cultivation result shows that the Burkholderia K4-1 has better environmental adaptability.
TABLE 2 influence of Potassium-solubilizing Agents on the growth of Chinese little greens
Test group Shanghai provinceChinese little greens plant height (cm) Fresh weight of Shanghai chicken feather dish (g/basin) Increase yield compared with CK
CK 8.82±0.19a 11.79±0.52a -
K4-1 12.61±0.57b 15.83±0.34b 34.27%
Example 3
500. mu.L of Burkholderia K4-1 bacterial suspension (bacterial concentration 10)8) Inoculated into a 100mL liquid medium (sucrose 1g, (NH)4)2SO4 0.1g、Na2HPO4·12H2O 1g、MgSO4·7H2O 0.075g、CaCO30.05g, potassium feldspar powder 0.2g, distilled water 100mL, pH 7.9, sterilization at 121 ℃ for 20min) in a 250mL conical flask, three parallel flasks were made, and no inoculation control was set. Performing shake culture at 28 deg.C and 200r/min for 6d, diluting the fermentation broth, spreading on solid potassium-decomposing culture medium, culturing in 28 deg.C constant temperature incubator for 3d, counting colony number, and measuring viable bacteria number in the fermentation broth to 6.44 × 109cfu/mL。
Example 4
500. mu.L of Burkholderia K4-1 bacterial suspension (bacterial concentration 10)8) Inoculated into a 100mL liquid medium (sucrose 0.5g, (NH)4)2SO4 0.1g、Na2HPO4·12H2O 1g、MgSO4·7H2O 0.075g、CaCO30.05g, potassium feldspar powder 0.2g, distilled water 100mL, pH 7.9, sterilization at 121 ℃ 20min) in 250mL Erlenmeyer flasks, three replicates were made, with no inoculation control. Performing shake culture at 28 deg.C and 200r/min for 6d, diluting the fermentation broth, spreading on solid potassium-decomposing culture medium, culturing in 28 deg.C constant temperature incubator for 3d, counting colony number, and measuring viable bacteria number in the fermentation broth to 5.69 × 109cfu/mL。
Example 5
500. mu.L of Burkholderia K4-1 bacterial suspension (bacterial concentration 10)8) Inoculated into a 100mL liquid medium (sucrose 1g, (NH)4)2SO4 0.05g、Na2HPO4·12H2O 1g、MgSO4·7H2O 0.075g、CaCO30.05g, potassium feldspar powder 0.2g, distilled water 100mL, pH 7.9, sterilization at 121 ℃ for 20min) in a 250mL conical flask, three parallel flasks were made, and no inoculation control was set. Performing shake culture at 28 deg.C and 200r/min for 6d, diluting the fermentation broth, spreading on solid potassium-decomposing culture medium, culturing in 28 deg.C constant temperature incubator for 3d, counting colony number, and measuring viable bacteria number in the fermentation broth to 5.05 × 109cfu/mL。
Example 6
500. mu.L of Burkholderia K4-1 bacterial suspension (bacterial concentration 10)8) Inoculated into a 100mL liquid medium (sucrose 1g, (NH)4)2SO4 0.1g、Na2HPO4·12H2O 0.75g、MgSO4·7H2O 0.075g、CaCO30.05g, potassium feldspar powder 0.2g, distilled water 100mL, pH 7.9, sterilization at 121 ℃ for 20min) in a 250mL conical flask, three parallel flasks were made, and no inoculation control was set. Performing shake culture at 28 deg.C and 200r/min for 6d, diluting the fermentation broth, spreading on solid potassium-decomposing culture medium, culturing in 28 deg.C constant temperature incubator for 3d, counting colony number, and measuring viable bacteria number in the fermentation broth to 3.33 × 109cfu/mL。
Example 7
500. mu.L of Burkholderia K4-1 bacterial suspension (bacterial concentration 10)8) Inoculated into a 100mL liquid medium (sucrose 1g, (NH)4)2SO4 0.1g、Na2HPO4·12H2O 1.25g、MgSO4·7H2O 0.075g、CaCO30.05g, potassium feldspar powder 0.2g, distilled water 100mL, pH 7.9, sterilization at 121 ℃ for 20min) in a 250mL conical flask, three parallel flasks were made, and no inoculation control was set. Performing shake culture at 28 deg.C and 200r/min for 6d, diluting the fermentation broth, spreading on solid potassium-decomposing culture medium, culturing in 28 deg.C constant temperature incubator for 3d, counting colony number, and measuring viable bacteria number in the fermentation broth to 4.97 × 109cfu/mL。
Example 8
500. mu.L of Burkholderia K4-1 bacterial suspension (bacterial concentration 10)8) Inoculated into a 100mL liquid medium (sucrose 1g, (NH)4)2SO4 0.1g、Na2HPO4·12H2O 1g、MgSO4·7H2O 1.25g、CaCO30.05g, potassium feldspar powder 0.2g, distilled water 100mL, pH 7.9, sterilized at 121 ℃ for 20min) were put into a 250mL Erlenmeyer flask, three replicates were made, and no inoculation control was set. Performing shake culture at 28 deg.C and 200r/min for 6d, diluting the fermentation broth, spreading on solid potassium-decomposing culture medium, culturing in 28 deg.C constant temperature incubator for 3d, counting colony number, and measuring viable bacteria number in the fermentation broth to 5.46 × 109cfu/mL。
Example 9
Taking 500 mu L of Burkholderia K4-1 bacterial suspension (the thallus concentration is 10)8) Inoculated into a 100mL liquid medium (sucrose 1g, (NH)4)2SO4 0.1g、Na2HPO4·12H2O 1g、MgSO4·7H2O 0.075g、CaCO30.1g potassium feldspar powder 0.2g, 100mL distilled water, pH 7.9, sterilization at 121 ℃ for 20min) in a 250mL conical flask, three parallel flasks were made, and no inoculation control was set. Performing shake culture at 28 deg.C and 200r/min for 6d, diluting the fermentation broth, spreading on solid potassium-decomposing culture medium, culturing in 28 deg.C constant temperature incubator for 3d, counting colony number, and measuring viable bacteria number in the fermentation broth to 6.18 × 109cfu/mL。
Example 10
500. mu.L of Burkholderia K4-1 bacterial suspension (bacterial concentration 10)8) Inoculated into a 100mL liquid medium (sucrose 1g, (NH)4)2SO4 0.1g、Na2HPO4·12H2O 1g、MgSO4·7H2O 0.075g、CaCO30.05g, potassium feldspar powder 0.1g, distilled water 100mL, pH 7.9, sterilization at 121 ℃ for 20min) in a 250mL conical flask, three parallel flasks were made, and no inoculation control was set. Performing shake culture at 28 deg.C and 200r/min for 6d, diluting the fermentation broth, spreading on solid potassium-decomposing culture medium, culturing in 28 deg.C constant temperature incubator for 3d, counting colony number, and measuring viable bacteria number in the fermentation broth to 6.08 × 109cfu/mL。
The separated Burkholderia K4-1 has better environmental adaptability, after the Burkholderia K4-1 bacterial suspension is applied, the plant growth can be obviously promoted by additional fertilization in the same soil environment, and the overground fresh weight and the plant height of the vegetable cultivated in the next year are increased by more than 30 percent compared with those of a control group. The burkholderia K4-1 bacterial suspension is applied in experimental soil, and the content of effective potassium in the soil is increased (the amplification is 9.6%) after 2 months.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. The potassium bacteria are classified and named as Burkholderia K4-1, and the preservation number is CCTCC M2020187.
2. A potassium-solubilizing bacterium agent for soil, comprising the potassium-solubilizing bacterium of claim 1.
3. The potassium bacteria inoculant according to claim 2, wherein the potassium bacteria inoculant is prepared by the following steps:
activating the potassium bacteria, inoculating the potassium bacteria on an activation culture medium, putting the activated potassium bacteria into a constant-temperature incubator, and culturing the activated potassium bacteria for 3-4 days at a constant temperature of 28 ℃;
and selecting strains on the activation culture medium, expanding and inoculating the strains to a potassium-solubilizing culture medium, culturing for 3-4 days at a constant temperature of 28 ℃, and washing the lawn to prepare a bacterial suspension, namely the potassium-solubilizing bacterial agent.
4. The potassium bacteria inoculant according to claim 2, wherein the potassium bacteria inoculant is prepared by the following steps:
activating the potassium bacteria, inoculating the potassium bacteria on an activation culture medium, putting the activated potassium bacteria into a constant-temperature incubator, and culturing the activated potassium bacteria for 3-4 days at a constant temperature of 28 ℃;
and selecting strains on the activation culture medium, inoculating the strains in a fermentation culture medium, carrying out shake culture at 28 ℃ for 6-7 days to obtain a fermentation liquid, centrifuging, collecting precipitates, and carrying out water resuspension to obtain the potassium bacteria decomposing agent.
5. The potassium bacteria inoculant according to claim 3 or 4, wherein the potassium bacteria slant medium is preserved.
6. The potassium bacteria dissolving agent as claimed in claim 4, wherein the centrifugation and collection of the precipitate comprises centrifugation at 5000-6000 r/min for 8-10 min to collect the precipitate.
7. The potassium bacteria inoculant according to claim 3 or 4, wherein the activation medium is Alexander bauov solid medium.
8. The potassium bacteria dissolving agent as claimed in claim 4, wherein the fermentation medium comprises the following components in parts by weight: 5-10 g of cane sugar and (NH)4)2SO4 0.5~1g、Na2HPO4·12H2O 7.5~12.5g、MgSO4·7H2O 0.75~1.25g、CaCO30.5-1 g of potassium feldspar powder, 1-2 g of potassium feldspar powder and 1000mL of distilled water, adjusting the pH value of the culture medium to 7.5-7.9, and sterilizing at 121 ℃ for 20min for later use.
9. The potassium bacteria inoculant according to claim 8, wherein the potassium feldspar powder is washed with deionized water for 3-5 times and then naturally dried in the shade.
10. Use of the potassium solubilizing bacteria of claim 1 or the potassium solubilizing bacteria agent for soil of any one of claims 2 to 9 in soil potassium solubilization.
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