CN117247867A

CN117247867A - Phosphorus-dissolving mosaic bacteria capable of degrading seaweed residues and application thereof

Info

Publication number: CN117247867A
Application number: CN202311235873.6A
Authority: CN
Inventors: 曾祥难; 韦建玉; 彭博; 杨祝军; 谢冰悦; 梁永进; 余育光; 卢炳泽; 成武洋
Original assignee: China Tobacco Guangxi Industrial Co Ltd
Current assignee: China Tobacco Guangxi Industrial Co Ltd
Priority date: 2023-09-22
Filing date: 2023-09-22
Publication date: 2023-12-19

Abstract

The invention relates to phosphorus-dissolving mosaic bacteria capable of degrading seaweed residues and application thereof, and belongs to the technical field of soil conditioning. The invention provides a phosphorus-dissolving mosaic bacterium (Massilia phosphatilytica) SD-1, and the preservation number of the phosphorus-dissolving mosaic bacterium SD-1 is CGMCC No.19791. The phosphorus-dissolving mosaic bacteria SD-1 can secrete a plurality of seaweed degrading enzymes, has short enzyme production fermentation time and strong capability of converting and utilizing seaweed residues, can extract residual organic nutrients in the seaweed residues by using the seaweed extracted waste residues as raw materials through a microbial degradation method to produce the biofertilizer capable of providing organic nutrient elements required by plant growth, thereby opening up a way for the application of the seaweed residues in agriculture for waste utilization. Meanwhile, the phosphorus-dissolving mosaic bacteria SD-1 can dissolve phosphate ores to generate soluble phosphorus so as to provide inorganic nutrient elements required for plant growth, and has the effect of improving soil.

Description

Phosphorus-dissolving mosaic bacteria capable of degrading seaweed residues and application thereof

Technical Field

The invention relates to phosphorus-dissolving mosaic bacteria capable of degrading seaweed residues and application thereof, and belongs to the technical field of soil conditioning.

Background

The sea algae resources in China are rich, and the algae with economic value are large-scale species such as brown algae, red algae, green algae, a small amount of blue algae and the like. To date, seaweeds have demonstrated great development potential and economic value in the fields and areas of foods, marine drugs, animal feeds, organic fertilizers, cosmetics, bioenergy, and the like. In the processing and utilizing process of the seaweed, the nutrients in the seaweed cannot be completely extracted due to the limitation of the existing process conditions, and the rest part remains in the waste seaweed residues. The seaweed residues contain substances such as alginic acid, mannitol, betaine, chitosan, polyphenol, alginate fiber, pectin, protein, lignin and the like besides partial moisture, and have rich nutrition. Therefore, how to further process and utilize the seaweed residues, and to exert the value to the greatest extent, becomes an increasingly concerned problem.

The seaweed lyase can degrade seaweed residues by decomposing alginate, alginic acid, algal polysaccharide, seaweed gel and the like. Seaweed lyase is classified into intracellular enzyme and extracellular enzyme according to enzyme production position, and classified into endo-enzyme and exo-enzyme according to enzyme cleavage mode. The seaweed lyase has wide sources and various types, and at present, nearly 100 seaweed lyases are separated, identified, cloned and purified from different species such as marine bacteria, terrestrial bacteria, marine mollusks and algae. Among them, marine bacteria are the most important source of seaweed lyase, and bacteria isolated from rotten kelp by marine Vibrio (Vibrio sp.) and Pseudoalteromonas (Pseudoalteromonas sp.) are capable of secreting seaweed salt lyase. However, these marine algae-derived bacteria that secrete algae lyase are difficult to adapt to the soil environment, which limits their development of application in agriculture.

Disclosure of Invention

In order to solve the problems, the invention provides a phosphorus-dissolving mosaic bacterium (Massilia phosphatilytica) SD-1, wherein the phosphorus-dissolving mosaic bacterium SD-1 is preserved in China general microbiological culture collection center (CGMCC) No.19791, and the preservation date is 2020, 05 and 09.

The phosphorus-dissolving mosaic bacteria SD-1 is derived from a multi-year tobacco planting soil sample of an ink test base of a tobacco institute of China academy of agricultural sciences in Qingdao, shandong province, the 16S rDNA sequence of the strain is shown as SEQ ID NO.1, and the sequence obtained by sequencing is subjected to nucleic acid sequence comparison, so that the result shows that the strain is the phosphorus-dissolving mosaic bacteria.

The invention also provides a phosphorus dissolving method, which comprises the following steps: inoculating the phosphorus-dissolving mosaic bacteria SD-1 into a culture system containing a poorly soluble inorganic phosphorus compound for culture, so as to convert the poorly soluble inorganic phosphorus compound in the culture system into soluble phosphorus. Phosphate solubilizing action, also known as phosphate solubilizing action. Refers to the process of converting organic phosphorus compounds in soil into Phosphate (POT) or converting poorly soluble phosphorus in soil into soluble phosphorus under the action of microorganisms. Can increase the content of available phosphorus in soil, and is beneficial to plant growth. The action of dissolving phosphorus in soil is divided into two types, enzymolysis type and acidolysis type. The former means that soil microorganisms gradually degrade organic phosphorus compounds such as nucleic acids, lecithin and phytin by using an enzyme system thereof, and finally convert into phosphate. The latter means that the solubility of calcium phosphate or apatite in the soil is increased by the dissolution of acidic metabolites of certain microorganisms, such as various organic acids and carbonic acid, sulfuric acid or nitric acid, etc. The effect of dissolving phosphorus (dissolving phosphorus) in soil is mainly dependent on the type and quantity of the matrix and ecological factors such as ventilation, moisture and soil pH, which influence activities of microorganisms involved in dissolving phosphorus.

In one embodiment of the present invention, the temperature of the culture is 25 to 40 ℃.

The invention also provides a method for degrading seaweed residues, which comprises the following steps: inoculating the phosphorus-dissolving mosaic bacteria SD-1 into a culture system containing seaweed residues for culture so as to degrade the seaweed residues in the culture system.

The invention also provides application of the phosphorus-dissolving mosaic bacteria SD-1 or the phosphorus-dissolving method in phosphorus dissolution.

The invention also provides application of the phosphorus-dissolving mosaic bacteria SD-1 or the method for degrading seaweed residues in degrading the seaweed residues.

The invention also provides application of the phosphorus-dissolving mosaic bacteria in preparing products, which is characterized in that the products have any one of the following functions:

(a) Dissolving phosphorus;

and/or, (b) degrading algae residue.

In one embodiment of the invention, the product comprises a soil conditioner and/or a biofertilizer.

The invention also provides a product, which contains the phosphorus-dissolving mosaic bacteria SD-1.

In one embodiment of the invention, the product has the functions as shown in any one of the following:

(a) Dissolving phosphorus;

and/or, (b) degrading algae residue.

The technical scheme of the invention has the following advantages:

the invention provides a phosphorus-dissolving mosaic bacterium (Massilia phosphatilytica) SD-1, and the preservation number of the phosphorus-dissolving mosaic bacterium SD-1 is CGMCC No.19791. The phosphorus-dissolving mosaic bacteria SD-1 can secrete a plurality of seaweed degrading enzymes, has short enzyme production fermentation time and strong capability of converting and utilizing seaweed residues, can extract residual organic nutrients in the seaweed residues by using the seaweed extracted waste residues as raw materials through a microbial degradation method to produce the biofertilizer capable of providing organic nutrient elements required by plant growth, thereby opening up a way for the application of the seaweed residues in agriculture for waste utilization. Meanwhile, the phosphorus-dissolving mosaic bacteria SD-1 can dissolve phosphate ores to generate soluble phosphorus so as to provide inorganic nutrient elements required for plant growth, and has the effect of improving soil. In addition, the phosphorus-dissolving mosaic bacteria SD-1 is derived from farmland soil, is different from strains derived from marine environment, can be directly applied to agricultural production, and has stronger resistance and adaptability to soil environment. In conclusion, the phosphorus-dissolving mosaic bacteria SD-1 can play an important role in relieving stress resistance of crops, improving soil, preventing and controlling soil insects and the like.

Preservation of biological materials

The phosphorus-dissolving mosaic bacteria (Massilia phosphatilytica) SD-1, which is named Massilia phosphatilytica in taxonomy, is preserved in China general microbiological culture Collection center (CGMCC) No.19791 in the year 2020, and has a preservation address of North Chen West Lu No.1 in the Korean region of Beijing city, wherein the preservation address is the China general microbiological culture Collection center of China, and the year 09 of the China is the year 05.

Drawings

Fig. 1: schematic representation of colonies.

Fig. 2: transparent circles observations (preliminary confirmation of strain degradability).

Fig. 3: schematic representation of purified colonies.

Fig. 4: transparent circles observations (reconfirmation of strain degradability).

Fig. 5: phylogenetic tree constructed by using 16S gene.

Fig. 6: phylogenetic tree of housekeeping genes.

Fig. 7: transcriptome sequencing wien plots. In FIG. 7, SD-A is the result of measurement of cells cultured in an alginate inorganic salt medium, and SD-G is the result of measurement of cells cultured in a glucose inorganic salt medium.

Fig. 8: transcriptome sequencing up-and down-regulated volcanic patterns. In fig. 8, the abscissa represents fold change values, i.e., FC values, of differences in gene expression between two groups of samples. The ordinate is a statistical test value of the variation in gene expression amount, i.e., p-value. The higher the p value, the more obvious the expression difference, and the numerical values of the abscissa and the ordinate are subjected to logarithmic treatment. Each dot in the figure represents a specific gene, the red dot represents a significantly up-regulated gene, the green dot represents a significantly down-regulated gene, and the gray dot is a non-significantly differential gene. After mapping all genes, it can be known that the left dot is a gene whose expression difference is down-regulated, the right dot is a gene whose expression difference is up-regulated, and the expression difference is more remarkable as the two and upper dots are located.

Detailed Description

The following examples are provided for a better understanding of the present invention and are not limited to the preferred embodiments described herein, but are not intended to limit the scope of the invention, any product which is the same or similar to the present invention, whether in light of the present teachings or in combination with other prior art features, falls within the scope of the present invention.

The following examples do not identify specific experimental procedures or conditions, which may be followed by procedures or conditions of conventional experimental procedures described in the literature in this field. The reagents or apparatus used were conventional reagent products commercially available without the manufacturer's knowledge.

Experimental example 1: obtaining, identifying and preserving phosphorus-dissolving mosaic bacteria SD-1

The method comprises the following specific steps:

soil collection and enrichment of degrading sodium alginate strains: the tobacco planting soil of the tobacco research institute, i.e. the ink test base of the national academy of agricultural sciences, qingdao, shandong is collected as a separation source sample. Firstly, air-drying and sieving the collected soil at room temperature (25 ℃) to remove sundries such as big stones, then taking 1kg of soil, adding sodium alginate powder accounting for 1% of the total mass of the soil, mixing, putting into a flowerpot, pouring tap water, and incubating for four weeks at room temperature.

Strain screening: weighing 10g of incubation soil, dissolving in a triangular flask filled with 90mL of sterile water, and shaking thoroughly to obtain 10 ^-1 A diluted soil suspension. Take 10 ^-1 Diluting the soil suspension to another triangular flask containing 90mL sterile water, shaking thoroughly to obtain 10 ^-2 A diluted soil suspension. And so on, get 10 in turn ^-3 、10 ^-4 、10 ^-5 、10 ^-6 A diluted soil suspension. Three dilutions of the soil suspension were pipetted onto screening plates (formula reference "Sawant s.s. for screening plates, salunke b.k., kim b.s. a rapid, positive, simple plate assay for detection of microbial alginate lyase activity. Enzyme and Microbial Technology (2015) 8-13"), specifically 0.5g/L peptone, 0.3g/L yeast extract, 2g/L sodium alginate, 2g/L ammonium sulfate, 1g/L potassium dihydrogen phosphate, 0.5g/L magnesium sulfate heptahydrate and 5g/L agar) and incubated in a 30 ℃ incubator for 48 hours to obtain colonies (see fig. 1). Bacterial colonies were picked up with sterilized toothpicks and backed up on a new screening plate, and incubated at 30℃for 48 hours, and then placed in a 4℃refrigerator.

Preliminary confirmation of the degradation ability of the strain: the transparent ring formed by adopting an iodine solution dyeing method is used for preliminarily determining the strain for degrading the sodium alginate, and the method specifically comprises the following steps: 1mL of the sterile solution was dropped onto a screening plate on which colonies were grown, the cells were peeled off from the screening plate by a glass coating rod, 5mL of gram iodine solution (product number A642190, purchased from Shanghai Biotechnology Co.) was dropped, and the transparent ring size was observed after 10 minutes of standing (see FIG. 2 for transparent ring observation results).

Purification of the strain and reconfirmation of the strain degradation ability: according to the size of the transparent ring on the original screening plate, picking the clone on the corresponding backup screening plate onto an ISP2 plate (the formula of the ISP2 plate is 4g/L yeast extract, 10g/L malt extract, 4g/L glucose and 20g/L agar), placing the clone in a 30 ℃ incubator for 48 hours, picking the newly grown monoclonal, inoculating the monoclonal into 5mL ISP2 liquid culture medium (the formula of the ISP2 liquid culture medium is used for removing agar on the basis of the ISP2 plate), and shake culturing for 24 hours at 30 ℃ and 200rpm to obtain a culture solution; during the culture, the growth of the strain was observed and found to be satisfactory. After spotting 5. Mu.L of the culture solution onto a screening plate and culturing the plate in an incubator at 30℃for 24 hours for a purified colony (purified colony is shown in FIG. 3), the transparent ring was again detected by the iodine staining method (see FIG. 4 for observation of transparent ring). The corresponding positive strain was stored in a-80℃refrigerator using an aqueous glycerol solution with a final concentration of 20% (v/v).

16S Gene identification assay: bacterial colony PCR amplification was performed on the isolated strains using bacterial universal primers 16S-27F (SEQ ID NO.2:5 '-AGAGTTTGATCMTGGCTCAG-3') and 16S-1492R (SEQ ID NO.3:5 '-TACGGYTACCTTGTTACGACTT-3'), specifically: dipping a bacterial colony by using a sterilized toothpick, and suspending in 50 mu L of sterilized redistilled water to obtain diluted bacterial liquid; mixing 2X Taq PCRMaster Mix. Mu.L, 22. Mu.L of sterile water, 1. Mu.L of each of the upstream and downstream primers (10. Mu.M) and 1. Mu.L of the diluted bacterial solution, centrifuging, and subjecting the resulting 50. Mu.L PCR system to PCR amplification (amplification conditions: 3min at 95℃for pre-denaturation, 30second at 95℃for denaturation, 30second at 55℃for annealing, 1.5min at 72℃for 30 cycles, and 10min at 72 ℃) for extension); the amplified product was submitted to Qingdao qing biotechnology company for detection and sequencing to obtain 16S rRNA of the isolated strain (the 16S rDNA sequence of the strain SD-1 is shown as SEQ ID NO. 1). After BLAST alignment analysis of the isolated strain 16S rRNA on NCBI (http:// www.ncbi.nlm.nih.gov /), it was again confirmed on the NZBioCloud website (https:// www.ezbiocloud.net /). The 16S sequences of the approximate strain were downloaded from NCBI database and phylogenetic tree was constructed using mega 6.0 (phylogenetic tree see FIG. 5). According to the characteristics of colony size, color, growth rate and the like, more than 90% of the strains identified from the screening plate are bacillus, wherein the strain SD-1 accounts for about 3% of the enriched strains. The strain SD-1 is further compared and identified by 16S, and the maximum similarity between the strain and the phosphorus-dissolving mosaic 12-OD1 is found, and the similarity reaches 99.6%, so that the strain is named as phosphorus-dissolving mosaic (Massilia phosphatilytica) SD-1. The phosphorus-dissolving mosaic bacteria SD-1 is preserved in China general microbiological culture Collection center (CGMCC) No.19791, and the preservation date is 2023, 04 and 27.

Experimental example 2: performance verification of phosphorus-dissolving mosaic bacteria

The experimental procedure was as follows:

genome sequencing analysis of carbohydrate utilization enzyme-based Gene experiments

After shaking culture of phosphorus-dissolving mosaic bacteria SD-1 to logarithmic phase at 30 ℃ and 200rpm by using ISP2 liquid culture medium, the bacteria are centrifugally collected and delivered to Shanghai Mejie sequencing company. The genome sketch of the bacterium is constructed, and the genome size of the phosphorus-dissolving mosaic bacterium SD-1 is found to be 7,311,277bp, 92 Scaffold are assembled, the GC content is 66.03%, 6411 proteins are encoded, 82 tRNA's are contained, and 1 rRNA is contained. The assembly conditions were as follows: the total frame length is Large Scaf Bases (bp) 7,311,277bp, and the total frame number is 92; the number of the large frames is 75, and the longest frame Largest Scaf Len (bp) is 554,169bp; scaf N50 (bp) was 316,925bp,Scaf N90 (bp) and 61,074bp, sequencing depth 219.64 fold.

The analysis result of the carbohydrate degrading enzyme system of the phosphorus-dissolving mosaic bacteria SD-1 is as follows: there are 159 glycoside hydrolases GH families, 72 glycosyltransferases, 68 carbohydrate esterases, 26 units with lignin degrading activity, 6 carbohydrate binding domains, and 15 other PL family polysaccharide hydrolases. The secretion system predicts that only two type I secretion systems, 18 type II secretion systems, 25 type VI secretion systems are found, and that no type IIII, IV, V secretion systems are found. Type III secretion systems are important plant virulence effector secretion systems and therefore have no potential for plant pathogenicity.

The phylogenetic tree was constructed according to whole genome sequencing, 19 strains closest to the species level were selected based on 31 housekeeping genes (dnaG, frr, infC, nusA, pgk, pyrG, rplA, rplB, rplC, rplD, rplE, rplF, rplK, rplL, rplM, rplN, rplP, rplS, rplT, rpmA, rpoB, rpsB, rpsC, rpsE, rpsI, rpsJ, rpsK, rpsM, rpsS, smpB, tsf) by comparison with a local database, and the phylogenetic tree was constructed by MEGA 6.0 software selection NJ (Neighbor-Joining) method (phylogenetic tree see fig. 6), showing that the relationship between phosphorus-dissolving mosaic SD-1 and malodorous mosaic was recent. The Marseilles putida can generate dimethyl disulfide, and can kill soil nematodes, soil pathogenic fungi and soil insects (see reference Massiia putida sp.nov., a dimethyl disulfide-producing bacterium isolated from wolfram mine tailing Int J Syst Evol Microbiol. 2016), so that the screened phosphorus-dissolving Marseilles putida SD-1 has great potential in the aspect of preventing and treating underground diseases.

Transcriptome sequencing comparative analysis degradation enzyme series experiment

Shaking culture of phosphorus-dissolving mosaic bacteria SD-1 to OD (OD) at 30 ℃ and 200rpm by using ISP2 liquid culture medium ₆₀₀ After=3.0, the cells were collected by centrifugation; respectively adding thallus into algin inorganic salt culture medium (namely inorganic salt ammonium salt culture medium containing 1g/L sodium alginate, the inorganic salt ammonium salt culture medium is purchased from Qingdao sea Bo biotechnology company, product number HB 8761), the formula of the inorganic salt ammonium salt culture medium is that potassium dihydrogen phosphate 3.0g/L, magnesium sulfate heptahydrate 0.1g/L, dipotassium hydrogen phosphate 1.0g/L, anhydrous calcium chloride 0.01g/L, disodium edetate 0.01g/L, ammonium nitrate 2.0gg/L and yeast powder 0.8 g/L) and glucose inorganic salt culture medium (namely inorganic salt ammonium salt culture medium containing 1g/L glucose) to OD ₆₀₀ After =0.5, shake-cultured at 30 ℃ for two hours at 200rpm, the cells were collected by centrifugation and submitted to Shanghai Meiji sequencing company, and the sequencing analysis results are shown in FIGS. 7 to 8 and Table 1.

In the quality control analysis of transcriptome sequencing original data of phosphorus-dissolving mosaic bacteria SD-1 in an algin inorganic salt culture medium and a glucose inorganic salt culture medium, the ratio Q20 and the ratio Q30 respectively reach 96% and 90%, the clear reads respectively reach 2911 ten thousand and 3200 ten thousand, and the clear bases respectively reach 3910Mb and 4137Mb.

Transcriptome sequencing of phosphorus-dissolving mosaic bacteria SD-1 in algin inorganic salt culture medium totally obtains the Total number of Total Reads of 29111506, 28245619 compared with genome, the ratio of the Total Reads to the genome of the genome is 97.03%, and the independent comparison rate of the Total Reads is 96.25%. Transcriptome sequencing in glucose mineral medium yielded a Total of 32002274 Total Reads of 30791265 aligned to the genome of the strain, with a alignment of 96.22% and a single alignment of 95.50%.

15 polysaccharide hydrolases in the phosphorus-dissolving mosaic bacteria SD-1 genome, eight of which are highly expressed in alginic acid culture medium. The PL9 family (mainly encoding pectin lyase, exogalacturonase, endorhamnogalacturonase, thiopeptidan lyase) has three genes highly expressed, genes 2913, 2922 and 3351, log2fc 3.25, 5.45 and 3.16 respectively. PL5 family has gene 4551 (log 2FC 3.58); the PL4-1 family (endorhamnogalacturonase) has gene 2136 (log 2FC 1.76); the PL22 family (galactono-lyase) has gene 3194 (log 2FC 2.22); the expression level of the PL18 family gene 1035 is highest, log2FC reaches 12.02, and the carbohydrate-active enzyme database CAZy database query finds that the PL18 family protein encoding algin lyase is mostly derived from marine bacteria Pseudomonas and Alternomonas, which probably is that degrading enzymes derived from marine bacteria are transferred into farmland microorganisms through a strategy of horizontal gene transfer. Except for the PL5 family gene 4551, which was predicted to be alginate lyase, the remaining genes were putative proteins.

TABLE 1 high expression polysaccharide lyase system in phosphorus-dissolving mosaic SD-1

Gene numbering	GH family	Log2FC	Encoded proteins
				gene1035	PL18	12.02	hypothetical protein(SEQ ID NO.4)
gene2136	PL4_1	1.76	hypothetical protein
				gene2913	PL9	3.25	hypothetical protein(SEQ ID NO.5)
gene2922	PL9	5.45	hypothetical protein(SEQ ID NO.6)
				gene3194	PL22	2.22	hypothetical protein(SEQ ID NO.7)
gene3351	PL9	3.16	hypothetical protein(SEQ ID NO.8)
				gene3656	PL11	0.73	rhamnogalacturonan lyase
gene4551	PL5	3.58	alginate lyase(SEQ ID NO.9)

Carbon source utilization experiment of Strain

The experiment of the utilization of each single carbohydrate was carried out by adding 0.5g/L of carbon source to the medium. The formulation of the medium used for the carbohydrate utilization experiments was as follows: 1.0g/L ammonium nitrate, 1.0g/L potassium dihydrogen phosphate, 1.0g/L dipotassium hydrogen phosphate, 0.2g/L magnesium sulfate heptahydrate, 0.02g/L calcium chloride, 0.05g/L ferric chloride, 0.8g/L yeast extract, and 5g/L carbon source. Single colonies of phosphorus-dissolving mosaic bacteria SD-1 are selected from a screening plate by using an inoculating needle and inoculated into 100mL of a culture medium used in a carbohydrate utilization experiment, and shake culture is carried out at 30 ℃ and 200rpm for two hours to determine OD ₆₀₀ Absorbance. It was found that phosphomosaic SD-1 can utilize lactose, starch, pectin, carboxymethyl cellulose, galacturonic acid, sodium potassium tartrate, and sodium citrate, but not fucose, mannitol, and chitosan. Meanwhile, it was found that phosphorus-dissolving mosaic bacteria SD-1 can grow on ISP2 and R2A media, but cannot grow on LB media, can grow at 10-40 ℃ and pH 5.0-8.0 (the optimal growth temperature is 28-30 ℃ and the optimal growth pH is 7.0-7.5), can generate white opaque colonies on ISP2 after 48 hours of culture at 30 ℃, and like actinomycetes, the colonies are tightly combined with solid media and have diameters of 2-3 mm.

API identification experiment

Single bacterial colony of phosphorus-dissolving mosaic bacteria SD-1 is picked from a screening flat plate by an inoculating needle to 0.9% (w/v, g/100 mL) of sterilizing physiological saline, carefully ground and vibrated to make the bacterial cells uniform, a tube part or a cup part is filled with bacterial suspension according to the application instruction of an API kit, part of small holes are covered with mineral oil to form an anaerobic environment, a culture box is covered, and incubation is carried out for 24 hours. It was found that phosphomosaic SD-1 was able to produce acids using lactose, peptone, starch, tyrosine, tween-20, tween-80, pectin, carboxymethyl cellulose, galacturonic acid, potassium sodium tartrate, but not using fucose, mannitol, chitosan, adipic acid, arabinose, glucose, malic acid, maltose, mannose, phenylacetic acid, potassium gluconate and trisodium citrate, but not using decanoic acid, mannitol and N-acetylglucosamine; nitrate reduction and oxidase were positive, whereas indole formation was not found; the enzyme activities of beta-glucosidase, protease and beta-galactosidase were positive, while the enzyme activities of arginine dihydrogenase and urease were negative.

Phosphorus dissolution ability experiment

Experiments were performed using PVK solid medium and PVK liquid medium (PVK liquid medium reference "Nautiyal CS. An efficient microbiological growth medium for screening phosphate solubilizing microorganic sms. FEMS Microbiol Lett 1999;170:265-270", formulation 10g/L glucose, 0.5g/L ammonium sulfate, 0.2g/L sodium chloride, 0.2g/L potassium chloride, 0.1g/L magnesium sulfate heptahydrate, 0.002g/L manganese sulfate monohydrate, 0.002g/L ferric sulfate heptahydrate and 5g/L basic calcium phosphate; solid medium was added 20g/L agar based on liquid medium). And (3) picking single bacterial colonies of the phosphorus-dissolving mosaic bacteria SD-1 from the screening plate by using an inoculating needle, inoculating the single bacterial colonies into an ISP2 liquid culture medium, and shake culturing at 30 ℃ and 200rpm until the OD600 = 3.0 to obtain bacterial liquid. After streaking the bacterial liquid on PVK solid culture medium and culturing the bacterial liquid in a 30 ℃ incubator for 48 hours, colony formation on the PVK solid culture medium is obviously observed, and the control escherichia coli cannot grow. After inoculating the bacterial liquid to PVK liquid medium with an inoculum size of 1% (v/v) and shake culturing at 30℃and 200rpm for 48 hours, the bacterial liquid was centrifuged (12,000 g,5 min), and the supernatant was subjected to detection of the content of soluble phosphorus by the method of blue molybdate (see "Murphy J, riley JP. A modified single solution method for the determination of phosphate in natural waters. Anal Chim Acta1962; 27:31-36"), whereby it was detected that the content of soluble phosphorus was 40. Mu.g/L.

Seaweed residue degradation and utilization test

Experiments were performed using an inorganic salt ammonium salt medium added with 2.5g/L seaweed residue (dry weight) as a degradation medium (seaweed residue was supplied by Qingdao Mingyu seaweed Co., ltd., water content was 80%, inorganic salt ammonium salt medium was purchased from Qingdao sea Bo biotechnology Co., product No. HB8761, and the formula of inorganic salt ammonium salt medium was 3.0g/L potassium dihydrogen phosphate, 0.1g/L magnesium sulfate heptahydrate, 1.0g/L dipotassium hydrogen phosphate, 0.01g/L calcium chloride anhydrous and 0.01g/L disodium ethylenediamine tetraacetate). And (3) picking single bacterial colonies of the phosphorus-dissolving mosaic bacteria SD-1 from the screening plate by using an inoculating needle, inoculating the single bacterial colonies into an ISP2 liquid culture medium, and shake culturing at 30 ℃ and 200rpm until the OD600 = 3.0 to obtain bacterial liquid. Inoculating the bacterial liquid into degradation culture medium with an inoculum size of 1% (v/v), shake culturing at 30deg.C and 200rpm for one week, filtering and weighing with 40 mesh sieve (pore diameter of 0.45 mm), and collecting supernatant to obtain seaweed liquid fertilizer. The reduction amount of dry matters before and after the culture of the seaweed residues is used as an index of the extraction rate, and the phosphorus-dissolving mosaic bacteria SD-1 can be calculated to degrade 95% of the seaweed residues in a week of the culture time.

It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the invention.

Claims

1. The phosphorus-dissolving mosaic bacteria (Massilia phosphatilytica) is characterized in that the phosphorus-dissolving mosaic bacteria are preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of 19791.

2. A method of dissolving phosphorus, the method comprising: inoculating the phosphorus-soluble mosaic bacteria of claim 1 into a culture system containing a poorly soluble inorganic phosphorus compound for culture, so as to convert the poorly soluble inorganic phosphorus compound in the culture system into soluble phosphorus.

3. A method of degrading algae residue, the method comprising: inoculating the phosphorus-dissolved mosaic bacteria of claim 1 into a culture system containing seaweed residues for culture, so as to degrade the seaweed residues in the culture system.

4. Use of the phosphorus-dissolving mosaic bacteria of claim 1 or the phosphorus-dissolving method of claim 2 in dissolving phosphorus.

5. Use of the phosphorus-dissolving mosaic bacteria of claim 1 or the method for degrading algae residue of claim 3 for degrading algae residue.

6. Use of a phosphorus-dissolving mosaic bacterium according to claim 1 for the preparation of a product, wherein said product has any one of the following functions:

(a) Dissolving phosphorus;

and/or, (b) degrading algae residue.

7. Use according to claim 6, wherein the product comprises a soil conditioner and/or a biofertilizer.

8. A product comprising the phosphorus-dissolving mosaic of claim 1.

9. A product according to claim 8, wherein the product has the functions as shown in any one of the following:

(a) Dissolving phosphorus;

and/or, (b) degrading algae residue.

10. The product according to claim 8 or 9, wherein the product comprises a soil conditioner and/or a biofertilizer.