Sinomicrobium strain SM8 and application thereof in salt-tolerant growth promotion
A technology of Microbacteria sinensis and Microbacteria, which is applied in microorganism-based methods, applications, microorganisms, etc., can solve the problems of poor adaptability and poor effect of related microorganisms, and achieve the effects of promoting crop growth, simple production and improved germination rate.
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Embodiment 1
[0030] Example 1 Isolation, screening, purification and preservation of Microbacterium sinensis SM8 CGMCC No.15757 strain
[0031] The rhizosphere soil samples of halophytes in Xinjiang Heshuo area were collected, and microorganisms were isolated and purified by gradient dilution method. Weigh 10g of the soil sample, put it into 100ml of sterile water (with glass beads added), shake at 200rpm, 30°C for 20min, to fully disperse the sample. After serial dilution, dilute with sterile water to prepare 10 -2 -10 -5 sample suspension diluent. Take 100 μL of the dilutions of each concentration, apply the conventional coating method on the TSA solid medium plate containing 2% NaCl, and place it in a constant temperature incubator at 30°C for cultivation. After 48 hours, after colonies grow on the plate, pick a single colony on the plate for purification and culture. For the purified single colonies ( figure 1 ), transferred to the slant of TSA medium containing 2% NaCl and stored...
Embodiment 2
[0032] Example 2 Identification of Microbacterium sinensis SM8 CGMCC No.15757 strain
[0033] Sequence determination and analysis of 16S rDNA of a new strain of Microbacterium sinensis CGMCC No.15757:
[0034] (1) Extraction of PCR template DNA:
[0035] The purified strain IS7 was inoculated into 2% NaCl TSA medium, cultured on a shaker at 30°C for 2 days, the bacteria were collected, and the total genomic DNA was extracted using a DNA extraction kit.
[0036] (2) PCR amplification
[0037] Using specific primers:
[0038] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3',
[0039] 1492R:5'-GGTTACCTTGTTACGACTT-3';
[0040] The total volume of the PCR reaction system was 25 μL, and the PCR amplification conditions were as follows: 94°C for 5 minutes; 94°C for 45s, 56°C for 45s, 72°C for 45s, 30 cycles; 72°C for 10 minutes.
[0041] (3) Sequence determination
[0042] The PCR amplified product was detected by electrophoresis and purified and then sequenced to obtain a 16S rDNA sequence wit...
Embodiment 3
[0045] Example 3 Detection of Salt Tolerance and Growth-promoting Ability of Microbacterium sinensis SM8 CGMCC No.15757 Strain
[0046] After adding 10% NaCl to the TSA medium, invert the plate and inoculate with Microbacterium sinensis SM8 CGMCC No.15757. Cultivate at 30°C, and observe the growth of the bacteria for 2-5 days.
[0047] Microbacterium sinensis SM8 CGMCC No.15757 was inoculated on a phosphate-solubilizing (inorganic and organic phosphate) bacterial culture medium at 30°C. After culturing for one week, observe the size of the transparent circle.
[0048] Microbacterium sinensis SM8 CGMCC No.15757 was inoculated in TSA liquid medium containing L-tryptophan (100mg / L) in 2% NaCl, cultured on a shaker (30°C, 180r / min) for 1d, and 50 μL of bacteria The suspension was dropped on a white ceramic plate, and an equal volume of Salkowski colorimetric solution (50mL 35% HClO 4 +1mL 0.5mol / L FeCl 3 ), and a mixed solution of 50 μL of uninoculated 2% TSA liquid medium and ...
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