Anaerobic river sediment degrading strain and application thereof
A technology of river sediment and strains, applied in the field of environmental microorganisms, can solve problems such as human health and ecological environment risks, and achieve the effects of reduction and harmlessness, high application value, and convenient operation
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Embodiment 1
[0030] Example 1: Isolation, screening and performance determination of anaerobic river sediment-degrading bacteria
[0031] The media and components used are as follows:
[0032] Liquid medium: glucose 5 g / L, tryptone 3 g / L, beef powder 2 g / L, NaCl 4 g / L, K 2 HPO 4 0.75 g / L, KH 2 PO 4 0.75 g / L, FeSO 4 ·7H 2 O 0.05 g / L, MgSO 4 ·7H 2 O 0.3 g / L, (NH 4 ) 2 SO 4 3 g / L, cysteine 0.25 g / L, natural pH.
[0033] Solid medium: glucose 5 g / L, tryptone 3 g / L, beef powder 2 g / L, NaCl 4 g / L, K 2 HPO 4 0.75 g / L, KH 2 PO 4 0.75 g / L, FeSO 4 ·7H 2 O 0.05 g / L, MgSO 4 ·7H 2 O 0.3 g / L, (NH 4 ) 2 SO 4 3 g / L, cysteine 0.25 g / L, 15 g / L agar powder, natural pH.
[0034] River sediment liquid medium: river sediment (the total solid TS content is 11%, and the VS content is 7.8% of TS, both are mass fractions). Add water to adjust the sediment TS content to 5%, and the pH value to 7.0.
[0035] River sediment solid medium: river sediment (TS content is 11%, VS content is 7...
Embodiment 2
[0042] Example 2: Biological Identification of Anaerobic River Sediment Degradation Strain YE6
[0043] Genomic DNA of strain YE6 was extracted and used as a template to amplify strain 16S rDNA with a pair of universal primers (27F, 1492R). The upstream primer is 27F (5'-AGAGTTTGATCCTGGCTCA-3'), and the downstream primer is 1492R (5'-GGTTACCTTGTTACGACTT-3').
[0044] The PCR reaction system (20 μL) is as follows: template DNA 0.5 μL, PCR Taqmix 10 μL, upstream and downstream primers 0.6 μL each, add ddH 2 O to 20 μL of reaction system.
[0045] PCR program: pre-denaturation at 94°C for 5 min, denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 1 min and 30 s, and a total of 30 cycles of the above, extension at 72°C for 10 min, and finally storage at 4°C.
[0046] The PCR product was purified and sequenced by Shanghai Jieli Biotechnology Co., Ltd., and the 16S rDNA sequence of the strain was obtained as shown in SEQ ID NO.1. The 16S rDNA sequence...
Embodiment 3
[0048] Embodiment 3: Preparation of liquid microbial agent of anaerobic river channel sediment degradation strain YE6
[0049] The YE6 pure strain was inoculated in 10 mL of liquid medium, cultured anaerobically at 37 °C for 24 h, and then inserted into the liquid medium of the next stage of expanded culture with an inoculum volume of 5% of the volume of the liquid medium. The liquid microbial agent of the strain can be obtained by carrying out multi-stage anaerobic expansion culture under the same conditions.
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