Rhizobium strain and application thereof
A rhizobia, XS-1 technology, applied in the direction of bacteria, biological water/sewage treatment, enzymes, etc., can solve the problem of cellulose hydrolysis efficiency drop
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Embodiment 1
[0053] 1. Isolation and identification of Rhizobium sp. XS-1CGMCC No.23976
[0054] Rhizobium sp. XS-1CGMCC No.23976 was isolated from a water sample in Suqian, Jiangsu in July 2020.
[0055]XS-1 is a Gram-negative bacterium, which forms a round regular yellow colony after being cultured on LB solid medium for 72 hours. The activated XS-1 was inoculated in LB liquid medium, and shaken at 30°C and 150rpm for 72h. Take 1ml of freshly cultured XS-1 bacterial solution and centrifuge at 4°C and 8000rpm for 5min, collect the bacteria in a 2ml centrifuge tube, and extract DNA using a DNA extraction kit. After electrophoresis detection, use the universal primers 8F and 1492R to perform PCR amplification using the above-mentioned extracted DNA as a template to detect 16S rDNA:
[0056] 8F: 5'-AGAGTTTGATCCTGGCTCA-3'
[0057] 1492R: 5'-GGTTACCTTGTTACGACTT-3'
[0058] The 16S rDNA of strain XS-1 has the nucleotide sequence of sequence 1 in the sequence listing. After comparison, XS-1...
Embodiment 2
[0072] Embodiment 2, the β-glucosidase activity of rhizobia XS-1
[0073] The preparation method of the solid medium A used in this example is as follows: LB liquid medium is used as the basal medium, and the medium obtained by adding aescin, ferric ammonium citrate and agar to the basal medium, in which seven The content of phylloside is 0.05% (m / v), the content of ferric ammonium citrate is 0.25% (m / v), and the content of agar is 1.5% (m / v).
[0074] The activated XS-1 was inoculated in LB liquid medium, and shaken at 30°C and 150rpm for 24h. Take 2 μl of fresh bacterial liquid and spot it on the above solid medium A, and culture it statically at 30°C for 48 hours. The result shows that there is an obvious black precipitation circle around the XS-1 colony, and the diameter ratio of the black precipitation circle to the colony is 17 / 8= 2.12, indicating that strain XS-1 has better β-glucosidase activity.
Embodiment 3
[0075] Example 3, functional detection of rhizobia XS-1 degrading phenol
[0076] The preparation method of the high-salt phenol medium used in this example is as follows (taking 1L as an example): 5g of peptone, 1g of yeast powder, 100g of sodium chloride, and 0.2g of phenol, dissolved in water to a volume of 1L, and sterilized at 115°C for 20 minutes .
[0077] The activated XS-1 was inoculated in LB liquid medium, and shaken at 30°C and 150rpm for 24h. Centrifuge the freshly cultured bacterial solution at 4°C and 8000 rpm for 5 minutes, collect the bacterial cells and suspend them with an equal volume of sterile water to form a bacterial suspension to obtain the XS-1 bacterial suspension.
[0078] Inoculation culture solution treatment: inoculate the XS-1 bacterial suspension in a 100mL Erlenmeyer flask containing 50mL high-salt phenol medium at a volume ratio of 5% (v / v), shake at 30°C and 150rpm for 3 days to obtain inoculation culture medium. Set up 3 repetitions, eac...
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