CN113549584B - High-potassium-decomposing paenibacillus mucilaginosus strain and application thereof - Google Patents
High-potassium-decomposing paenibacillus mucilaginosus strain and application thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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Abstract
The invention discloses a high-potassium-decomposing paenibacillus mucilaginosus strain, which is paenibacillus mucilaginosus GT018-JZ18112 and is preserved in China general microbiological culture Collection center in 2019, 5 months and 7 days, wherein the preservation address is No. 3 of Xilu No.1 of Xingyang district of Beijing city, and the preservation number is CGMCC No.17702. The strain has excellent potassium-decomposing capacity and can improve soil fertility.
Description
Technical Field
The invention relates to the technical field of agricultural microorganisms, in particular to a high-potassium-decomposing paenibacillus mucilaginosus strain and application thereof.
Background
Potassium is one of three elements of all plant nutrients, and plays an important role in growth and development of plants, formation of yield and quality. Tissues and organs with vigorous metabolic activity of plants contain more potassium, and more than sixty enzymes in plants participate in the activity of potassium. Potassium can affect the metabolism of nitrogen and carbohydrates, and affect the synthesis of proteins, starches and oils. Potassium plays an important role in the transportation of substances in plants, and the transportation tissues (phloem cells) of the plants contain more potassium (2%). The potassium-deficient plant has thinned mechanical tissue and cell wall, weak stem and easy lodging and is easy to be attacked by diseases and pests. Potassium affects physical properties of plant cell pectin substances, regulates osmotic pressure of cytoplasm, controls stomata opening and closing of leaves, and reduces unnecessary water loss. Potassium can change the quality of agricultural products, improve the sugar content of beet, sugarcane and berry, enhance the fragrance and combustibility of flue-cured tobacco, and improve the quality of fruits and vegetables.
The paenibacillus mucilaginosus is also called bacillus mucilaginosus, is an important functional bacterium in soil, can decompose original ecological minerals of aluminosilicate such as feldspar, mica and the like, enables insoluble K, P, si and the like in the soil to be converted into soluble substances for plant growth and utilization, can generate various bioactive substances to promote the plant growth, can improve the yield of crops, increases the stress resistance, and also increases the fertility of the soil.
At present, one of the main factors limiting agricultural yield worldwide is soil impoverishment and fertility reduction, and the supplement of nutrient elements necessary for the growth and development of crops becomes an essential important measure for obtaining stable and high yield. The soil is a natural potassium reservoir, but is mainly present in primary minerals containing potassium (mica and potassium feldspar) and cannot be absorbed and utilized by plants. The paenibacillus mucilaginosus can convert insoluble invalid potassium in potassium-containing minerals such as potassium feldspar and the like into soluble valid potassium, however, the existing paenibacillus mucilaginosus has weak potassium dissolving capacity and weak spore producing capacity, so that the application of the paenibacillus mucilaginosus as a potassium fertilizer is limited.
Therefore, how to provide paenibacillus mucilaginosus with high potassium-dissolving function and apply the paenibacillus mucilaginosus to agricultural production is a problem which needs to be solved urgently by the technical personnel in the field.
Disclosure of Invention
In view of the above, the invention provides a high potassium-decomposing paenibacillus mucilaginosus strain, wherein the strain is paenibacillus mucilaginosus GT018-JZ18112, and the strain has excellent potassium decomposing capability and can improve soil fertility.
In order to achieve the purpose, the invention adopts the following technical scheme:
the high potassium-decomposing paenibacillus mucilaginosus strain is paenibacillus mucilaginosus GT018-JZ18112 which is preserved in China general microbiological culture Collection Center (CCM) 5.7.2019 with the preservation address of No. 3 Siro-1 of Beijing Korean-Yang district, and the preservation number is CGMCC No.17702.
The technical effect achieved by the technical scheme is as follows: the paenibacillus mucilaginosus GT018-JZ18112 can decompose silicate, aluminosilicate and potassium-containing minerals in other ores, has the functions of dissolving phosphorus, releasing potassium and fixing nitrogen, and releases medium and trace elements such as soluble calcium, sulfur, magnesium, iron, zinc, molybdenum, manganese and the like, thereby not only improving the soil fertility, but also providing absorbable and utilizable comprehensive nutrient elements for crops. Meanwhile, organic acid, amino acid, polysaccharide and other substances which are beneficial to the absorption and utilization of plants can be generated in the growth and propagation processes. After the plant growth stimulant and various enzymes are propagated in soil, the plant growth stimulant and various enzymes are secreted to enhance the resistance of crops to some diseases and inhibit the growth of other pathogenic bacteria.
An application of high potassium-decomposing paenibacillus mucilaginosus strain in preparing a biological fertilizer.
A biological fertilizer comprises the Paenibacillus mucilaginosus GT018-JZ18112.
According to the technical scheme, compared with the prior art, the high-potassium-decomposing paenibacillus mucilaginosus strain is disclosed and provided, and the biofertilizer is prepared from the strain, so that the physicochemical property of soil can be effectively improved, the content of organic matters and quick-acting potassium in the soil can be increased, the conventional fertilization is reduced by 10% in the cucumber planting period, the content of the microbial inoculum of the paenibacillus mucilaginosus GT018-JZ18112 is 10 kg, the yield is increased by 425.6 kg/mu compared with the conventional fertilization, and the yield is increased by 10.27%; the conventional fertilization is reduced by 10% + 10 kilograms of paenibacillus mucilaginosus microbial inoculum (a commercial product), the yield is increased by 241.7 kg/mu, and the yield is increased by 5.59%; compared with the blank control, the yield is increased by 712.3 kg/mu, the yield is increased by 18.48 percent, and the yield increasing effect is extremely obvious.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The embodiment of the invention provides a screening method and application of a paenibacillus mucilaginosus strain GT018-JZ18112, the screened paenibacillus mucilaginosus strain GT018-JZ18112 is separated from wheat rhizosphere soil and serves as a starting strain, high-temperature ultraviolet mutagenesis is carried out, a flat potassium-resolving ring is used for selecting the strain, then verification is carried out through a shaking flask fermentation spore formation rate experiment, and potassium-resolving capability determination is carried out in soil, so that the high-potassium-resolving capability paenibacillus mucilaginosus GT018-JZ18112 is finally obtained, the strain is preserved in the common microorganism center of China Committee for culture Collection of microorganisms in 2019, 5 and 7 days, the preservation address is No. 3 of the Shangyang district of Beijing, the preservation number is CGMCC No.17702.
The embodiment of the invention also relates to the application of the paenibacillus mucilaginosus strain GT018-JZ18112 in preparing biological fertilizer, in particular to the application in preparing biological potassium fertilizer.
Example 1 Strain screening
A method for obtaining paenibacillus mucilaginosus GT018-JZ18112, which comprises the following steps:
1) Preparation of bacterial cells of hairstyle-developing strains
(11) Separating and screening out an initial strain from rhizosphere soil of Baoding wheat in Hebei river;
(12) The original strain is treated at 60 ℃, 80 ℃ and 100 ℃ for 20min and then inoculated into a potassium-dissolving culture medium, and cultured in a constant-temperature incubator at 32 ℃ for 48h for later use.
2) Screening of ultraviolet mutagenesis high potassium-decomposing performance bacterial strain
(21) Putting the strain prepared in the step 1 into 10ml of sterile water, and fully oscillating to disperse cells for later use;
(22) And (3) pouring the solid potassium decomposing culture medium into a flat plate, and solidifying for later use, wherein the solid potassium decomposing culture medium comprises the following components: sucrose 5-10%, na 2 HPO 4 1-2%, potassium feldspar powder 10-20%, (NH) 4 ) 2 SO 4 0.5-0.7％、MgSO 4 ·7H 2 O1-2％、CaCO 3 1-2%, yeast extract 0.5-0.7%, feCl 3 Trace agar 1.8-2%, pH7.0-7.2 (this culture method)The culture medium is a mutagenesis guide selection culture medium
(23) Ultraviolet ray mutagenesis treatment: and (3) putting the bacterial liquid to be used in the step (21) into a sterile culture dish (the diameter is 6 cm) with a magnetic stirring rod, and adding 3ml of bacterial liquid into each dish. The above culture dish was placed on a magnetic stirrer and irradiated for 2min and 5min, respectively. Performing gradient dilution on the irradiated bacterial suspension and the non-irradiated bacterial suspension, coating the diluted bacterial suspension on a (22) flat plate, and then performing light-proof culture at 32 ℃ for 48h;
(24) Obvious potassium-dissolving transparent rings appear on a solid potassium-dissolving culture medium flat plate, and three strains with wide transparent rings are discovered through repeated experiments.
But the potassium-dissolving transparent ring cannot be used as the only index for judging the potassium-dissolving capacity of the bacteria, so the potassium-dissolving capacity is further determined: the potassium-decomposing capacity of the three strains is quantitatively measured by adopting a flame photometry method, and the three strains are inoculated to a liquid culture medium which takes potassium feldspar as a unique potassium source. The culture solution is treated by a hydrogen oxide digestion method, and potassium-decomposing capacity is measured, so that the release amount of potassium in the liquid culture medium is in positive correlation with a strain potassium-decomposing ring size control, and then the paenibacillus mucilaginosus GT018-JZ18112 with high potassium-decomposing capacity is screened out, the strain can decompose 234mg of effective potassium higher than the control from 100g of potassium feldspar powder under pure culture conditions, and the increase amplitude is 70.3%. The strain is preserved in China general microbiological culture Collection center (CGMCC) in 2019, 5 and 7 days, wherein the preservation address is No. 3 of Xilu No.1 of Beijing university and Chaoyang district, and the preservation number is CGMCC No.17702.
Identification of strains
The obtained Paenibacillus mucilaginosus (paenibacillus mucinagonosus) GT018-JZ18112 has the following characteristics:
the cells are rod-shaped, gram-negative, and have a size of 0.9-1.1. Mu. M.times.3.0-5.0. Mu.m. The spores are oval, thick, medium to partial medium, and the blastocysts are micro-expanded. Intracellular production of poly-beta-hydroxybutyrate (PHB) granules.
The growth of the strain is good on a silicate bacteria culture medium, thick capsules are generated, the bacterial colony is circular, bulged and transparent, the surface is smooth and glossy, the texture is sticky and rich in elasticity, and the strain is not easy to pick up.
Positive reaction: contacting with an enzyme.
Negative reaction: an oxidase; lecithin enzyme; anaerobic growth; the nutrient gravy grows.
16s rDNA sequence analysis: extracting genome DNA from thallus, amplifying 16s rDNA by using primer, and sequencing PCR product. The tested 16s rDNA sequence is subjected to BLAST comparison with the sequence of related species in a GenBank database after proofreading and splicing, and the result shows that the 16s rDNA sequence of the strain has 100 percent of sequence homology with the paenibacillus mucilaginosus (paenibacillus mucoginosus).
16s rDNA sequence:
<xnotran> CGGCTTCGGGTGTTGTAAACTCTCGTGGTGTGACGGGCGGTGTGTACAAGACCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCAATTCCGACTTCATGCAGGCGAGTTGCAGCCTGCAATCCGAACTGAGACCGGCTTCTAAGGATTCGCTCCATCTCGCGACTTCGCTTCCCGTTGTACCGGCCATTGTAGTACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCACTCTAGAGTGCCCAACTCAATGCTGGCAACTAAAGTCAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCACCTCTGTCCCGAAGGAGCCCTATCTCTAGGGCTTTCAGAGGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACGACATACTCCACTGCTTGTGCGGGTCCCCGTCAATTCCTTTGAGTTTCACTCTTGCGAGCGTACTCCCCAGGCGGAGTGCTTATTGTGTTTACTTCGGCACCAAGGGTATCGAAACCCCTAACACCTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGCGCCTCAGCGTCAGTTACAGTCCAGAAAGCCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCGCTTTCCTCTCCTGCACTCAAGTCTTCCAGTTTCCGGTGCGAACCGGGGTTGAGCCCCGGGCTTAAACACCAGACTTAAAAGACCGCCTGCGCGCGCTTTACGCCCAATAATTCCGGACAACGCTTGCCCCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGGGCTTTCTTCTCAGGTACCGTCATTCGCAGAGCAGTTACTCTCCACGACATTCTTCCCTGGCAACAGAGCTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGGCTTGCGCCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTCGCCTTGGTGGGCCGTTACCCCGCCAACTAGCTAATGCGCCGCAGGCCCATCCGTAAGCCACAGGTTGCCCCGTGTTTCATGATTCCGGCATGCACCGAAACCAGCTATCCGGTCTTAGCTACCGTTTCCGGTAGTTATCCCGATCTTACAGGCAGGTTGCCTACGTGTACTCACCCGTCCGCCGCTAAGCACCGAA, SEQ ID NO.1 . </xnotran>
Example 2 field application test
Cucumber field application test of paenibacillus mucilaginosus:
selecting a test field: the test field is moist soil, the texture is medium soil, the fertility is medium, the previous crop is tomato, soil taking analysis is carried out on the plough layer before the test, the organic matter is 13.6g/kg, the total nitrogen is 1.18g/kg, the available phosphorus is 11.5mg/kg, the quick-acting potassium is 71mg/kg, the pH value is 8.1, and the test crop is No. 106 of cucumber Zhongnong;
test protocol and method: the experiment was designed with 4 treatments, 4 replicates, and a completely randomized block design. Cell area 6.00m is multiplied by 5.00m =30.00m 2 。
Treatment 1: and (5) applying fertilizers conventionally.
And (3) treatment 2: the conventional fertilization is reduced by 10% + the paenibacillus mucilaginosus JZ18112 microbial inoculum by 10 kilograms.
Preparation of paenibacillus mucilaginosus GT018-JZ18112 microbial agent: the paenibacillus mucilaginosus powder and the diatom shale are mixed into 5 hundred million spores/g powder.
And (3) treatment: 10% reduction of conventional fertilization + 10 kg of Paenibacillus mucilaginosus microbial agent (commercially available product)
And (4) treatment: blank control, no fertilizer was applied.
Other field management measures are consistent.
Test results and analysis
Effect of different treatments on biological traits of cucumber
Test results show that the root system of the cucumber plant is developed in a cell where the test fertilizer is additionally applied. The plants are strong, the growth is vigorous, the leaves are fat and green; ineffective flowers and deformed cucumbers are reduced, and the disease and pest resistance is strong; the plant height, stem thickness, leaf length, single plant fruiting and single fruit weight are all increased compared with other treatments, which shows that the product can promote the growth of cucumber plants and lay a certain foundation for high yield.
The effect of different treatments on cucumber yield is shown in table 1.
TABLE 1 statistics of cell yields
As can be seen from the table 2, the conventional fertilization of the cucumber by the paenibacillus mucilaginosus of the invention as a biological fertilizer reduces 10% + the bacterial manure of the paenibacillus mucilaginosus GT018-JZ18112 increases the yield by 425.6 kg/mu and increases the yield by 10.27% compared with the conventional fertilization; compared with the conventional fertilization, the fertilizer is reduced by 10% + 10 kilograms of paenibacillus mucilaginosus microbial agent (a commercial product), the yield is increased by 241.7 kg/mu, and the yield is increased by 5.59%; compared with the blank control, the yield is increased by 712.3 kg/mu, the yield is increased by 18.48 percent, and the yield increasing effect is extremely obvious.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Sequence listing
<110> research and promotion center of microbial fertilizer technology
<120> high-potassium-decomposing paenibacillus mucilaginosus strain and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1354
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
cggcttcggg tgttgtaaac tctcgtggtg tgacgggcgg tgtgtacaag acccgggaac 60
gtattcaccg cggcatgctg atccgcgatt actagcaatt ccgacttcat gcaggcgagt 120
tgcagcctgc aatccgaact gagaccggct tctaaggatt cgctccatct cgcgacttcg 180
cttcccgttg taccggccat tgtagtacgt gtgtagccca ggtcataagg ggcatgatga 240
tttgacgtca tccccacctt cctccggttt gtcaccggca gtcactctag agtgcccaac 300
tcaatgctgg caactaaagt caagggttgc gctcgttgcg ggacttaacc caacatctca 360
cgacacgagc tgacgacaac catgcaccac ctgtcacctc tgtcccgaag gagccctatc 420
tctagggctt tcagagggat gtcaagacct ggtaaggttc ttcgcgttgc ttcgaattaa 480
acgacatact ccactgcttg tgcgggtccc cgtcaattcc tttgagtttc actcttgcga 540
gcgtactccc caggcggagt gcttattgtg tttacttcgg caccaagggt atcgaaaccc 600
ctaacaccta gcactcatcg tttacggcgt ggactaccag ggtatctaat cctgtttgct 660
ccccacgctt tcgcgcctca gcgtcagtta cagtccagaa agccgccttc gccactggtg 720
ttcctccaca tctctacgca tttcaccgct acacgtggaa ttccgctttc ctctcctgca 780
ctcaagtctt ccagtttccg gtgcgaaccg gggttgagcc ccgggcttaa acaccagact 840
taaaagaccg cctgcgcgcg ctttacgccc aataattccg gacaacgctt gccccctacg 900
tattaccgcg gctgctggca cgtagttagc cggggctttc ttctcaggta ccgtcattcg 960
cagagcagtt actctccacg acattcttcc ctggcaacag agctttacga tccgaaaacc 1020
ttcatcactc acgcggcgtt gctccgtcag gcttgcgccc attgcggaag attccctact 1080
gctgcctccc gtaggagtct gggccgtgtc tcagtcccag tgtggccgat caccctctca 1140
ggtcggctac gcatcgtcgc cttggtgggc cgttaccccg ccaactagct aatgcgccgc 1200
aggcccatcc gtaagccaca ggttgccccg tgtttcatga ttccggcatg caccgaaacc 1260
agctatccgg tcttagctac cgtttccggt agttatcccg atcttacagg caggttgcct 1320
acgtgtactc acccgtccgc cgctaagcac cgaa 1354
Claims (3)
1. The high-potassium-decomposing paenibacillus mucilaginosus strain is characterized by being paenibacillus mucilaginosus GT018-JZ18112 which is preserved in China general microbiological culture Collection center on 5-7.2019, wherein the preservation address is No. 3 of Xilu No.1 of Beijing Korean district, and the preservation number is CGMCC No.17702.
2. The use of a paenibacillus mucilaginosus strain of claim 1 for the preparation of biofertilizer.
3. A biofertilizer comprising the paenibacillus mucilaginosus GT018-JZ18112 according to claim 1.
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| CN114908025B (en) * | 2022-06-23 | 2023-04-28 | 河北冀微生物技术有限公司 | Paenibacillus mucilaginosus HB-02 strain and application thereof in promoting crop growth |
| CN117660236B (en) * | 2023-11-30 | 2024-06-14 | 湖北茂盛生物有限公司 | Potassium-dissolving gel-like bacillus mucilaginosus MSSW02 and application thereof |
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Application publication date: 20211026 Assignee: Baoding Duowei Fertilizer Sales Co.,Ltd. Assignor: MICROBIAL FERTILIZER TECHNOLOGY RESEARCH AND EXTENSION CENTER Contract record no.: X2023980031579 Denomination of invention: A strain of Bacillus gelatinosus with high potassium solubility and its application Granted publication date: 20221118 License type: Common License Record date: 20230118 |