Red-spotted stonecrop root lead-resistant strain Providence bacterium, screening method and application of red-spotted stonecrop root lead-resistant strain Providence bacterium
A Providence, strain technology, applied in microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve problems such as human health hazards, soil pollution, disruption of normal ecosystem functions, etc.
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Embodiment 1
[0028] Example 1 Isolation of Sedum rhizosphere lead-resistant bacterial strain Providencia sp.BPb7
[0029] Materials and Methods
[0030] 1. Medium
[0031] Bacterial culture medium: beef extract 3g, peptone 10g, NaCl 5g, distilled water 1000mL, pH7.0.
[0032] Prepare the screening medium: the basic medium is 1000ml of bacterial culture medium, add 10mL of trace element solution and vitamin solution, sterilize at 121°C for 20min, cool to about 70°C, add lead solution mother liquor, so that the lead content in the medium is 1000mg· L -1 ; Culture at 37°C.
[0033] The trace element solution is: MnSO 4 0.01g, ZnSO 4 0.05g, H 3 BO 3 0.01g, CaCl 2 0.01g, dilute to 1L, and store at 4°C in the dark. The vitamin solution is: creatine 0.025g, ascorbic acid 0.025g, riboflavin 0.025g, citric acid 0.02g, dilute to 1L, and store in the dark at 4°C. Lead solution mother liquor: Pb(CH 3 COOH) 2 ·3H 2 O formulated as Pb 2+ The concentration is 200mg·mL -1 The mother liquor ...
Embodiment 2
[0036] The strain identification of embodiment 2 bacterial strain BPb7
[0037] 1. Genomic DNA extraction
[0038] The above-mentioned strain BPb7 was mass-cultured according to conventional technical means, and then its genomic DNA was obtained.
[0039] 2. Identification method of 16S rDNA of strain BPb7
[0040] 2.1 PCR amplification, sequencing and phylogenetic tree construction of 16S rDNA of strain BPb7
[0041] 2.1. PCR amplification of 116S rDNA gene sequence
[0042] The primers at both ends of the amplified 16S rDNA gene sequence are universal primers: forward primer BSF8 / 20: 5'-AGAGTTTGATCCTGGCTCAG-3' (SEQ ID NO: 1) and reverse primer BSR1541 / 20: 5'-AAGGAGGTGATCCAGCCGCA-3' (SEQ ID NO: 2). The PCR reaction system was 50 μL, and the reaction conditions were denaturation at 94°C for 5 minutes; followed by 35 cycle reactions: denaturation at 94°C for 45 s, annealing at 50°C for 45 s, and extension at 72°C for 90 s; then extension at 72°C for 10 min, and finally stor...
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