CN117106614A - Rhizosphere bacterium pseudomonas solanacearum YIM B08402, microbial agent and application thereof - Google Patents
Rhizosphere bacterium pseudomonas solanacearum YIM B08402, microbial agent and application thereof Download PDFInfo
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 26
- 241000873303 rhizosphere bacterium Species 0.000 title claims abstract description 7
- 241000589771 Ralstonia solanacearum Species 0.000 title claims description 19
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 66
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims abstract description 50
- 229910052698 phosphorus Inorganic materials 0.000 claims abstract description 50
- 239000011574 phosphorus Substances 0.000 claims abstract description 50
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims abstract description 40
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 33
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 23
- 150000007965 phenolic acids Chemical class 0.000 claims abstract description 21
- 239000003617 indole-3-acetic acid Substances 0.000 claims abstract description 20
- 239000002689 soil Substances 0.000 claims abstract description 20
- 235000007688 Lycopersicon esculentum Nutrition 0.000 claims abstract description 18
- 241000589516 Pseudomonas Species 0.000 claims abstract description 16
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 10
- 239000011780 sodium chloride Substances 0.000 claims abstract description 5
- 239000000589 Siderophore Substances 0.000 claims description 20
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- 238000004519 manufacturing process Methods 0.000 claims description 13
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- 238000000354 decomposition reaction Methods 0.000 claims description 11
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 11
- 239000010452 phosphate Substances 0.000 claims description 11
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 10
- 239000011591 potassium Substances 0.000 claims description 10
- 229910052700 potassium Inorganic materials 0.000 claims description 10
- FJKROLUGYXJWQN-UHFFFAOYSA-N 4-hydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 claims description 8
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 claims description 8
- 230000000593 degrading effect Effects 0.000 claims description 8
- 230000003381 solubilizing effect Effects 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 5
- 229940090248 4-hydroxybenzoic acid Drugs 0.000 claims description 4
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- 230000003248 secreting effect Effects 0.000 claims description 4
- 230000028327 secretion Effects 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 235000015278 beef Nutrition 0.000 claims description 3
- 239000000787 lecithin Substances 0.000 claims description 3
- 235000010445 lecithin Nutrition 0.000 claims description 3
- 229940067606 lecithin Drugs 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 238000004090 dissolution Methods 0.000 claims description 2
- 235000009048 phenolic acids Nutrition 0.000 claims 2
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- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 abstract description 4
- 229910052742 iron Inorganic materials 0.000 abstract description 2
- 230000009466 transformation Effects 0.000 abstract description 2
- 241000227653 Lycopersicon Species 0.000 abstract 3
- 230000008092 positive effect Effects 0.000 abstract 1
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- 241000894006 Bacteria Species 0.000 description 15
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- 238000011282 treatment Methods 0.000 description 10
- 238000012360 testing method Methods 0.000 description 9
- 230000015556 catabolic process Effects 0.000 description 8
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- 230000003301 hydrolyzing effect Effects 0.000 description 3
- 229910017053 inorganic salt Inorganic materials 0.000 description 3
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- 238000012163 sequencing technique Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N Iron oxide Chemical compound [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
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- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 2
- 238000004382 potting Methods 0.000 description 2
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- 238000011144 upstream manufacturing Methods 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 235000011612 Pinus armandii Nutrition 0.000 description 1
- 240000000793 Pinus armandii Species 0.000 description 1
- 241000589157 Rhizobiales Species 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- WYWFMUBFNXLFJK-UHFFFAOYSA-N [Mo].[Sb] Chemical compound [Mo].[Sb] WYWFMUBFNXLFJK-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
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- 238000005904 alkaline hydrolysis reaction Methods 0.000 description 1
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 1
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- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
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- 235000010958 polyglycerol polyricinoleate Nutrition 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/27—Pseudomonas
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The application discloses a rhizosphere bacterium Pseudomonas qinghaiensis YIM B08402 and application thereof, and the rhizosphere bacterium is classified and named as Pseudomonas qinghaiensisPseudomonas qingdaonensis) YIM B08402, deposited with China center for type culture Collection, accession number: cctccc NO: m2023153. The application performs the biological characteristics of the strain YIM B08402The detection shows that the strain can grow in the environment of 0-6% NaCl, has the functions of dissolving inorganic phosphorus, dissolving organic phosphorus, fixing nitrogen and producing iron carrier, secretes indoleacetic acid and degrades phenolic acid, and the rhizosphere microbial environment is more suitable for growth of tomato plants after the strain is prepared into a biological microbial agent and applied to the tomato plants, and nutrition transformation and supply have positive effects on promoting the growth of the tomatoes and improving the soil, so that the strain has good application prospect.
Description
Technical Field
The application belongs to the technical field of microorganisms, and particularly relates to rhizosphere bacteria YIM B08402 and application of a microbial agent mainly prepared from the rhizosphere bacteria YIM B08402.
Background
The high-speed development of agriculture enables people to select a faster and more convenient agricultural production mode, and the application of chemical fertilizer can effectively shorten the planting time and accelerate the accumulation of plant nutrient substances. Chemical fertilizers such as nitrogen fertilizers and phosphate fertilizers are the most common fertilizer types, are essential nutrient elements in the plant growth process, and are widely used in agricultural production, but long-time large-area application can cause adverse effects on soil, and environmental problems such as soil sample hardening, salinization, degradation and the like are accompanied by the problem that agricultural workers are searching for novel environment-friendly fertilizers with chemical effects similar to those of fertilizers. Biological fertilizers are increasingly attracting attention as a more environment-friendly and healthy fertilizer type, the main functional core sources are microorganisms, and the intensity and the quantity of the functions of the microorganisms directly determine the advantages and disadvantages of the microbial fertilizers.
Plant growth promoting bacteria (Plant growth promoting rhizobacteria, PGPR) survive on the rhizosphere or root surface of plants and can promote or regulate plant growth in some direct or indirect ways, while some of them also possess a class of beneficial microorganisms with biological control functions. The plant rhizosphere growth promoting bacteria have rich functions, including phosphate dissolving, potassium dissolving, nitrogen fixing, iron secretion, etc., can directly or indirectly promote the growth and development of plants, have excellent application prospect, screen and obtain multifunctional high-activity bacterial strains from the bacterial strains, and make microbial agents, thereby bringing new contributions to the nutrition transformation of plants and the benign change of soil environment.
Disclosure of Invention
The application aims to provide a rhizosphere bacterium pseudomonas qinghaiensis YIM B08402 and application thereof, which is used for screening a pseudomonas qinghaiensis strain and a microbial agent thereof from the rhizosphere soil of a fumelia mongolica, wherein the pseudomonas qinghaiensis strain has the functions of inorganic phosphorus decomposition, organic phosphorus decomposition, nitrogen fixation, siderophore production, indoleacetic acid secretion and phenolic acid degradation, can improve the soil environment, has obvious promotion effect on plant growth, and is applied to tomatoes.
The application aims at realizing the following technical scheme:
the application provides a pseudomonas qingdao strainPseudomonas qingdaonensis) YIM B08402, preserved in China Center for Type Culture Collection (CCTCC), with preservation address of university of Wuhan, china, preservation date of 2023, 2 months and 20 days, and preservation number of CCTCC NO: m2023153.
The application also provides application of the pseudomonas solanacearum YIM B08402 in inorganic phosphorus decomposition, organic phosphorus decomposition, nitrogen fixation, siderophore production, indoleacetic acid secretion and phenolic acid degradation.
The pseudomonas solanacearum has the functions of dissolving inorganic phosphorus, dissolving organic phosphorus, fixing nitrogen, producing siderophores, secreting indoleacetic acid and degrading phenolic acid, and can achieve positive promotion effect on plant growth when being prepared into microbial agents.
Further, the inorganic phosphorus is Ca 3 (PO 4 ) 2 The organic phosphorus is lecithin, and the phenolic acid is benzoic acid and p-hydroxybenzoic acid.
The application also provides an application of the pseudomonas solanacearum YIM B08402 in promoting plant growth.
Further, the plant is tomato.
Further, the biological microbial inoculum mainly prepared from pseudomonas solanacearum YIM B08402 can effectively improve root length, plant height, fresh weight and dry weight of planted tomatoes.
Further, the present application is carried out at 30 DEG CPseudomonas qingdaonensisYIM B08402 was still viable in the 6% NaCl environment.
Furthermore, the application also provides a microbial agent comprising the pseudomonas solanacearum YIM B08402.
The beneficial effects are that: compared with the prior art, the application has the following advantages:
the application separates, screens and identifies rhizosphere high-efficiency functional bacteria from rhizosphere soil samples of a natural protection area of a non-quantitative mountain country, researches the phosphorus dissolving effect, nitrogen fixing effect and siderophore capability, secretion of indoleacetic acid and degradation of phenolic acid of the rhizosphere high-efficiency functional bacteria to evaluate the capability of the functional bacteria to convert indissolvable phosphorus in soil into effective phosphorus which can be absorbed by plants, the capability of converting atmospheric nitrogen into ammonium nitrogen which can be directly utilized by the plants, the capability of degrading phenolic acid class autotoxic substances secreted by the plants and the capability of the bacterial strain to promote growth of indissolvable iron oxide and indoleacetic acid. The application shows that the application dissolves indissolvable phosphorus and secretes siderophores, can be used for promoting plant growth and increasing crop yield, improving agricultural productivity and promoting green development of ecological agriculture.
1) Rhizosphere bacteria YIM B08402 identified as Pseudomonas qinghaiensisPseudomonas qingdaonensis) Belongs to Pseudomonas Qingdao. Pseudomonas Qingdao is a new species of Pseudomonas which is considered to be a species of Pseudomonas by analyzing data based on phenotypes and 16SrRNA gene sequences by a heterogeneous taxonomy method in 2018 of Jae-Won Lee et alPseudomonas qingdaonensisJJ3. The pseudomonas solanacearum YIM B08402 has the advantages of better salt tolerance, organic phosphorus decomposition, inorganic phosphorus decomposition, nitrogen fixation, siderophore production, indoleacetic acid secretion and phenolic acid degradation, and has the development and application potential as a novel microbial strain resource.
2) The test result shows that the pseudomonas solanacearum YIM B08402 has the capabilities of fixing nitrogen, dissolving phosphorus, producing siderophores, secreting indoleacetic acid and degrading phenolic acid. A potting experiment shows that the microbial agent taking the pseudomonas solanacearum YIM B08402 as a core has a strong growth promoting effect on tomatoes, the plant height, the stem diameter, the dry fresh weight of overground parts, the root length and the dry fresh weight of the roots of the tomatoes are obviously improved, and the conventional eight items of measuring rhizosphere soil of the tomatoes also prove that the strain YIM B08402 has strong nutrition conversion capability and the pseudomonas solanacearum YIM B08402 has good nutrition conversion capability.
Drawings
FIG. 1 is a colony morphology of a strain on nutrient agar medium.
FIG. 2 shows the phosphate solubilizing ring of the strain on inorganic phosphorus solid medium.
FIG. 3 shows the phosphate solubilizing loop of the strain on the solid medium of organic phosphorus.
FIG. 4 is a nitrogen fixation circle of the strain on an Abbe (Asbhy) nitrogen-free medium.
FIG. 5 shows the color circle of the strain on CAS color solid medium for detecting siderophores.
FIG. 6 is a evolutionary tree of strain YIM B08402
Detailed Description
Preservation of organisms
The application provides pseudomonas solanacearumPseudomonas qingdaonensis) The rice is preserved in China center for type culture collection (China) for 2 months and 20 days in 2023, and has the address of eight paths 299 of Wuchang district of Wuhan, hubei province, university of Wuhan and the preservation number of CCTCC NO: m2023153.
The application will be further illustrated with reference to specific examples.
The plant material is tomato seedling, and the matrix is rhizosphere surface soil of Huilongcun in Panlong area of Kunming, yunnan province. The test medium is beef extract peptone (NA) medium, and inorganic phosphorus solid medium is used for determining dissolved Ca of strain 3 (PO 4 ) 2 The organic phosphorus solid culture medium is used for measuring the lecithin dissolving capacity of the strain, the ASS Bei Modan culture medium is used for qualitatively measuring the nitrogen fixing capacity, the CAS chromogenic solid culture medium is used for qualitatively measuring the function of the siderophore, quantitative experiments are carried out by using KB culture medium and Sackowcki's chromogenic reagent to prove the IAA secretion capacity, phenolic acid degrading detection is carried out by using phenolic acid inorganic salt liquid culture medium, excel 2016 software is adopted for data processing measured data, and SPSS software and Origin 2018 software are used for drawing for data statistics analysis.
Example 1
Bacterial YIM B08402 isolated from rhizosphere soil of Pinus armandi in the natural protection zone of mountain-free countries is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of: cctccc NO: m2023153.
1) And (3) observing the form and characteristics of the thallus and measuring physiological and biochemical indexes: the strain YIM B08402 was subjected to physiological and biochemical measurement by referring to "Berger's bacteria identification Manual" and "common bacteria System identification Manual", and colony morphology and characteristics were described.
Bacterial strain YIM B08402 had a nearly circular colony on NA medium, irregular edges, yellow color, raised middle of colony, moist and glossy (FIG. 1). The cells are rod-shaped, flagellum-shaped and gram-negative.
The strain YIM B08402 can grow in the environment of pH6.0-8.0,0-6% (w/v) NaCl and has a certain salt tolerance.
2) 16S rDNA gene sequence analysis and phylogenetic tree: the bacterial genomic DNA was amplified using a pair of universal primers (upstream primer 27F (5'-AGAGTTTGATCCTGGCTCAG-3'), downstream primer 1492R (5'-TACGGCTACCTTGTTACGACTT-3')) for bacterial 16SrDNA using the extracted bacterial total DNA as a template.
PCR reaction system: mix10mL, 1mL each of the upstream and downstream primers, 1mL of DNA template, and ddH 2 O7mL. PCR reaction conditions: pre-denaturing at 94 ℃ for 5min, denaturing at 94 ℃ for 30s, annealing at 55 ℃ for 30s, extending at 72 ℃ for 1min,33 cycles, extending at 72 ℃ for 10min, electrophoresing the obtained PCR product in 1% agarose gel, EB staining, and photographing under an ultraviolet lamp. The PCR products obtained were subjected to purification sequencing by Beijing engine technology company if there were bands. The gene sequence of the 16S rRNA was aligned with the sequences in Ez BioCloud and GenBank, and a phylogenetic tree was constructed with MEGA7 0.
The 16S rDNA amplified product of the strain YIM B08402 was detected by electrophoresis on a 1% agarose gel, resulting in a bright PCR band of interest around 1500bp and no other nonspecific bands. The target DNA fragment obtained by amplification of 16SrDNA was shown to be useful for sequencing. The 16S rDNA gene of the strain YIM B08402 obtained by sequencing has 1395 bases. The strain YIM B08402 and sequences in EzBioCloud and GenBank are subjected to gene sequence comparison, and sequence information of mode bacteria with highest homology and named are used for constructing a phylogenetic tree through MEGA7.0 software, and the strain YIM B08402 and Pseudomonas Qingdao are subjected to the following steps ofPseudomonas qingdaonensis) The JJ3 strains are clustered together, and the similarity reaches 100.00%. According to phylogenetic tree similarity analysis and combining morphology, physiological and biochemical characteristic comparison, the strain YIM B08402 is primarily identified as Pseudomonas qinghaiensisPseudomonas qingdaonensis)。
Example 2
This example is used to demonstrate the effect of rhizosphere bacteria YIM B08402 on dissolving inorganic phosphorus, dissolving organic phosphorus, fixing nitrogen, producing siderophores, secreting indoleacetic acid and degrading phenolic acid
1) And (3) inorganic phosphorus decomposition characteristic detection: the test strain was inoculated into an inorganic phosphorus medium by a 4-point inoculation method, each treatment was repeated 3 times, and placed in a 30℃incubator, and whether or not a transparent ring was produced around the colony was continuously observed. And the ratio between the diameter (D) of the transparent ring produced thereby and the colony diameter (D) was determined, and the D/D value of strain YIM B08402 was 2.316 (+ -0.159).
Quantitative detection of bacterial strain phosphorus dissolution: 300ml of LB strain seed solution was prepared, and the seed solution was aspirated into 100ml of inorganic phosphorus liquid medium in an amount of 1%, and the culture was repeated 3 times, and incubated at 180r/min and 30℃for 7d. Measuring pH value of supernatant after centrifuging culture solution daily, and measuring OD of supernatant daily by molybdenum-antimony anti-colorimetry 600 Values. The soluble phosphorus content of the strain was determined for 7 days according to the prepared phosphorus standard curve. The highest phosphate solubilizing amount of the strain YIM B08402 in the 1d is up to 878.95mg/L.
2) Detection of organophosphorus-degrading properties
The test strain was inoculated into an organophosphorus medium by a 4-point inoculation method, each treatment was repeated 3 times, and placed in a 30℃incubator, and whether or not a transparent ring was produced around the colony was continuously observed. And the ratio between the diameter (D) of the transparent ring produced thereby and the colony diameter (D) was determined, and the D/D value of strain YIM B08402 was 2.691 (+ -0.49).
3) Nitrogen fixation property detection
The test strain was inoculated into a nitrogen-free medium of Abbe (Asbhy) by 4-point inoculation, each treatment was repeated 3 times, and placed in an incubator at 30℃to continuously observe whether a transparent ring was produced around the colony. And the ratio between the diameter (D) of the transparent ring produced thereby and the colony diameter (D) was determined, and the D/D value of strain YIM B08402 was 2.595 (+ -0.884).
4) Siderophore production assay
The test strain was inoculated into a CAS chromogenic double-layer medium by a 4-point inoculation method, each treatment was repeated 3 times, and the culture was placed in a 30℃incubator, and whether a chromogenic ring was generated around the colony was continuously observed. And the ratio between the diameter (D) of the resulting color circle and the colony diameter (D) was measured, and the D/D value of the strain YIM B08402 was 2.91 (+ -0.57).
5) Quantitative IAA production assay
And (3) inoculating the activated pseudomonas solanacearum YIM B08402 into KB culture medium, shake culturing at 30 ℃ at 180 rpm/min for 24 h, absorbing 1mL fermentation liquor in a centrifuge tube under aseptic condition, rapidly mixing with 4mL Sackowcki's color reagent, standing at normal temperature for color development for 40min under dark condition, observing and recording color change, and if pink appears, determining that the strain is positive, thus indicating that the strain can secrete IAA.
After the strain is cultured in LB medium for 1d, seed liquid is prepared, the seed liquid is inoculated into LB liquid medium containing L-tryptophan with an inoculum size of 1 percent, the seed liquid is placed in a shaking table with a constant temperature of 30 ℃ for 180r/min for shaking culture, 4ml of supernatant liquid is sucked every day and mixed with 4ml of Sackowcki's color reagent, and after the seed liquid is kept stand for 40min in a dark environment, OD value is measured at a wavelength of 535 nm. Substituting the obtained absorbance value into a standard curve for calculation to obtain the content of the indoleacetic acid produced by the strain, and determining that the highest content of the indoleacetic acid produced by the strain YIM B08402 on day 3 is 24.57+/-0.14 mug/mL.
6) Qualitative detection of degraded phenolic acid
After the test strain is cultured in LB liquid medium for 1 day, the test strain is centrifugally resuspended to prepare seed liquid, and inoculated in 100ml of liquid medium containing 2.0mmol/L benzoic acid inorganic salt and 100ml of liquid medium containing 2.0mmol/L p-hydroxybenzoic acid inorganic salt with an inoculum size of 2 percent, after culturing for 48 hours, phenolic acid degradation liquid is taken out, sterile water is taken as blank control, the phenolic acid content is measured by adopting an ultraviolet spectrophotometry method, the absorbance of the phenolic acid degradation liquid is obviously higher than that of the control, and the strain YIM B08402 has the capability of degrading phenolic acid substances such as benzoic acid and p-hydroxybenzoic acid.
The determination shows that the rhizosphere bacteria YIM B08402 strain grows on inorganic phosphorus culture medium, organic phosphorus culture medium and Abbe (Asbhy) nitrogen-free culture medium to form transparent circles (figures 2, 3 and 4), red color circles appear in CAS color double-layer culture medium (figure 5), IAA qualitative detection appears pink, and absorbance in a phenolic acid degradation test is obviously higher than that of a control, and the results show that the strain has the capabilities of fixing nitrogen, dissolving inorganic phosphorus, dissolving organic phosphorus, fixing nitrogen, producing siderophore and producing indoleacetic acid.
Example 3
This example is for illustrating the growth promoting effect of microbial agents, mainly rhizosphere bacteria YIM B08402, on tomatoes.
Picking single colony with inoculating loop, shake culturing at 30deg.C and 180r/min for 3d in 500mL triangular flask containing 200mL NB culture medium, preparing microbial agent, applying the liquid bacterial solution to 7d planted tomato potted seedling, and applying 120mL bacterial agent (bacterial body amount 5.0X10) per plant 9 cfu/g), control group CK1 was inoculated with the same volume of sterile water, and control group CK2 was inoculated with the same volume of NB medium. After application, the tomato pot is placed in a greenhouse, and the plant height, stem diameter, dry fresh weight of the aerial parts, root length and dry fresh weight of the roots of the treatment group and the control group are measured 30d after the sterilization.
TABLE 1 Effect of post 30d inoculation strains on tomato growth
After the bacterial treatment, the plant height, the stem diameter, the dry fresh weight of the overground part, the root length and the dry fresh weight of the root are obviously improved compared with the control group CK 1. Compared with CK1 strain, the net growth of tomatoes in the post-treatment group for 30d is increased by 118%, the stem diameter is increased by 32%, the fresh weight of overground parts is increased by 528%, the dry weight of overground parts is increased by 477%, the root length is increased by 37%, the fresh weight of roots is increased by 413%, the dry weight of roots is increased by 747%, and the effect of effectively promoting plant growth of pseudomonas solanacearum YIM B08402 is demonstrated. There was a significant difference between the applied treatment and the control treatment (P < 0.05). The microbial agent mainly inoculated with rhizosphere bacteria YIM B08402 strain has remarkable promotion effect on tomato growth.
Example 4
This example is intended to illustrate the effect of the rhizosphere bacterium YIM B08402 on soil.
And collecting rhizosphere soil of the tomato plant when the potting experiment is finished, and performing conventional eight detection on the collected soil sample, wherein the detection comprises PH, organic carbon, total nitrogen, total phosphorus, total potassium, alkaline hydrolysis nitrogen, quick-acting phosphorus and effective potassium, and the benign change of the strain YIM B08402 to the soil environment is verified.
According to the physical and chemical indexes of soil of tomato rhizosphere, it can be obviously found that after the microbial agent taking the strain YIM B08402 as a main component is applied, the pH value and total phosphorus of three treatments are not obviously different, the total potassium and quick-acting potassium contents of the applied microbial agent are higher than those of two control groups, and the contents of organic matters, total nitrogen, total phosphorus, hydrolytic nitrogen and hydrolytic nitrogen are lower than those of the control groups.
By analyzing together with the tomato plant data in table 1, it can be presumed that, because the strain YIM B08401 has the functions of inorganic phosphorus decomposition, organic phosphorus decomposition and nitrogen fixation, the nutrient components in the soil are converted into available phosphorus and hydrolytic nitrogen which can be absorbed by plants, and are fully absorbed by tomato root systems, so that the organic matter content of the tomato plants is increased, the nutrient substances are accumulated, and the nitrogen and phosphorus content in the soil environment is reduced. Meanwhile, the increase of phosphorus and nitrogen under the influence of the growth promoting effect of the strain YIMB08401 is presumed, and under the positive interaction with potassium, the increase of potassium nutrition components is promoted, the total potassium content and the quick-acting potassium content of the soil are increased, and the obvious growth promoting effect of the strain YIMB08401 on the growth of tomatoes is proved.
Claims (14)
1. A rhizosphere bacterium is characterized in that the strain is pseudomonas solanacearumPseudomonas qingdaonensis) YIM B08402, deposited at China center for type culture Collection, university of Wuhan, china, with a date of deposit of 2023, 2 months and 20 days, with a deposit number of: cctccc NO: m2023153.
2. The application of pseudomonas solanacearum YIM B08402 in dissolving inorganic phosphorus, dissolving organic phosphorus and fixing nitrogen as claimed in claim 1.
3. The use according to claim 2, characterized in that the inorganic phosphorus is Ca 3 (PO 4 ) 2 The organic phosphorus is lecithin.
4. Use of pseudomonas solani YIM B08402 in siderophores as claimed in claim 1.
5. Use of pseudomonas solanacearum YIM B08402 in secretion of indoleacetic acid as claimed in claim 1.
6. Use of pseudomonas solanacearum YIM B08402 of claim 1 for degrading phenolic acids.
7. The use according to claim 6, wherein the phenolic acids are benzoic acid and p-hydroxybenzoic acid.
8. A microbial agent with the effects of inorganic phosphorus decomposition, organic phosphorus decomposition, nitrogen fixation, siderophore production and indoleacetic acid production, which is characterized by comprising pseudomonas solanacearum with the effects and a liquid carrier according to claim 1.
9. The microbial agent for decomposing inorganic phosphorus, organic phosphorus, nitrogen, siderophore and indoleacetic acid according to claim 8, wherein the living bacterial count of pseudomonas solanacearum in the microbial agent is more than 5.0x10 9 cfu/g。
10. The microbial agent with the effects of phosphate solubilizing, nitrogen fixation, siderophore production and indoleacetic acid production according to claim 8 or 9, wherein the liquid carrier is prepared from raw materials including peptone, beef extract powder, sodium chloride and distilled water.
11. The microbial agent with the effects of phosphate dissolution, nitrogen fixation, siderophore production and indoleacetic acid production as claimed in claim 8, wherein the mass ratio of peptone, beef extract powder, sodium chloride and water is 10:3:5:1000, the pH value is 7.4+/-0.2, and the microbial agent is sterilized for 20min at 121 ℃.
12. The method for preparing the microbial agent with the effects of phosphate solubilizing, nitrogen fixation, siderophore production and indoleacetic acid production according to any one of claims 8 to 11, which is characterized by comprising the following steps:
the strain YIM B08402 is inoculated into the liquid according to claim 10 and 11, and cultured for 3d at 30 ℃ and 180rpm, so as to obtain the microbial agent with the functions of phosphate dissolving, nitrogen fixing, siderophore producing and indoleacetic acid producing.
13. Use of pseudomonas solanacearum with phosphate and potassium solubilizing effect according to claim 1 or of a microbial agent with phosphate and nitrogen solubilizing, siderophore producing, indoleacetic acid secreting and phenolic acid degrading effects according to any one of claims 8-12 for improving soil and plant quality.
14. The use according to claim 13, wherein the plant is tomato.
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