Bacillus mucilaginosus and method for preparing bio-organic fertilizer through solid state fermentation of bacillus mucilaginosus

[0022]Example 1:

[0023]Isolation of pinacterus

[0024]Its separation process is usually the following steps:

[0025]The first step is to prepare Alexandrove culture base: its components include: NA2HPO4 2G, FECL3 0.005g, MgSO4 · 7h2O 0.5g, Caco3 0.1 g, sucrose 3g, potassium long stone powder (washed 5 times with deionized water) 1 g, distilled water 1000 mL, pH 7.0, agar 20g, and 20 min.

[0026]In the second step, the initial sieve of Potashus: Incorporal inoculation into the cultural substrate of Alexanderov culture, 10 days of 37 ° C constant temperature incubator, and there is no transparent ring formation on the medium. size. The diameter of the hydrolysis ring and colonies can be measured by a straight-foot, and the relative size of the strain is probabulous can be judged according to the diameter ratio of the hydrolysis ring to the colony. Pick the largest strain in the water and name it, and is named strain M-1.

[0027]Example 2:

[0028]16S rDNA sequence molecule identification of strain M-1

[0029]The strain QF-101 genomic DNA was extracted using CHELEX method, and 16S RDNA general primer 27f (5'-agagttgatcmtgctcAg-3 '), 1492R (5'-ggttacctTgTCGACTTTT-3'), 16S RDNA gene of strain QF-101 genomic DNA The long sequence was subjected to PCR amplification. The PCR product was subjected to 1% agarose gel electrophoresis, and the purpose of the SanPREP column DNA gel recovery kit (SANGONBIOTECH, ORDER No. B518131) was used to recover the destination DNA fragment. Sequence analysis of DNA fragments using 3730XL DNA Analyzer (Applied Biosystems), sequencing kits use Big Dyterminator V3.1 (Applied Biosystems).

[0030]The two-way sequencing results were spliced ​​using Seqman (DNASTAR Lasergene) to get a sequence of 16srRRNA gene close to the full length. Use eZBiocloud Identify Service (https://www.ezbiocloud.net/identify) and NCBI Nucleotide Blast (https://blast.ncbi.nlm.nih.gov/blast.cgi), the 16s rRNA gene sequence result is used with the GenBank database To compare, select a valid naming mode strain as a reference.

[0031]The strain M-1 was identified as a Bacillus Mucilaginosus. The morphological characteristics of the strain are: on the nitrogen-free solid medium, colony pilot, exhibit glass hemispherical, smooth, transparent, glue viscous, flexible; bacteria is rod, both ends are blunt, size It is 4 to 7 μm × 1 to 1.4 μm; after culturing 30 h in a starch medium, a large amount of spores, branches or near-end, elliptical, a size of about 1.5 to 1.7 μm × 2.8 ~ 3.4 μm are formed in the cells. .

[0032]Example 3:

[0033]Qualitative test of M-1 xacemic ability of strains

[0034]The culture solution of the cultured strain M-1 was 50 ml of 50 ml of 250 mL, and the fermentation liquid sterilized at 121 ° C for 30 min was blank. 12.5 g of potassium longlite powder, added to the culture solution and inactivated medium, each treatment for 3 parallel, cultured at 37 ° C, 160 rpm, 5th and 10th day, potassium tetrazoaborate The content of soluble potassium was determined by weight.

[0035]Table 1 strain M-1 解 potassium capacity qualitative test

[0036] 5 days of solution (mg / L) 10 days of solution (mg / L) Blank control 2.381 4.912 Strain M-1 30.735 58.975

[0037]The strain has been deposited in the normal microbiological center of China Microbial Square Safety, and the deposit number is CGMCC No. 19428, named Bacillus Mucilaginosus M-1. The bacterium has a strong solution of potassium potassium, and the ability to promote the ability of the culture solution can be 5d and 10d to reach 30.735 mg / l and 58.975 mg / L.

[0038]Bacillus M-1 16S rDNA sequence of this strain

[0039]Streptococcus M-1 16S rDNA sequence

[0040]AGAGTTTGATCCTGGCTCAGGACGACCGCTGGCGGCGTGCCTAATACAT GCAAGTCGAGCGGAGCACTTCGGTGCTTAGCGGCGGACGGGTGAGTAA CACGTAGGCAACCTGCCTGTAAGATCGGGATAACTACCGGAAACGGTA GCTAAGACCGGATAGCTGGTTTCGGTGCATGCCGGAATCATGAAACACG GGGCAACCTGTCGCTTACGGATGGGCCTGCGGCGCATTAGCTAGTTGGC GGGGTAATGGCCCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGC AGCAGTAGGGAATCTTCCGCAATGGGCGCAAGCCTGACGGAGCAACGC CGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGCCAGGGAA GAATGTCGTGGAGAGTAACTGCTCTGCGAATGACGGTACCTGAGAAGA AAGCCCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGGGC AAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGTCTTTT AAGTCTGGTGTTTAAGCCCGGGGCTCAACCCCGGTTCGCACCGGAAAC TGGAAGACTTGAGTGCAGGAGAGGAAAGCGGAATTCCACGTGTAGCGG TGAAATGCGTAGAGATCTGGAGGAACACCAGTGGCGAAGGCGGCTTTC TGGACTGTAACTGACGCTGAGGCGCGAAAACGTGGGGAGCAAACAGG ATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGGTGTTA GGGGTTTCGATACCCTTGGTGCCGAAGTAAACACAATAAGCACTCCGCC TGGGGAGTACGCTCGCAAGAGTGAAACTCAAAGGAATTGACGGGGACC CGCACAAGCAGTGGAGTATGTGGTTTAATTCGAAGCAACGCGAAGAAC CTTACCAGGTCTTGA CATCCCTCTGAAAACCCTAGAGATAGGGCCCTCC TTCGGGACAGAGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTC GTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGACTTTAGTTGCCAGCATTGAGTTGGGCACTCTAGAGTGACTGCCGGTGACAAACAG GAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGC TACACACGTACTACAATGGCCGGTACAACGGGAAGCGAAGTCGCGAGA TGGAGCGAATCCTTAGAAGCCGGTCTCAGTTCGGATTGCAGGCTGCAAC TCGCCTGCATGAAGTCGGAATTGCTAGTAATCGCGGATCAGCATACCGC GGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCACGAGA GTTTACAACACCCGAAGCCGGTGGGGTAACCCGTAAGGGAGCCAGCCG TCGAAGGTGGGGTAGATGAATGGGGTGAAGTCGTAACAAGGTAACC