An Antimony-resistant Bacteria nxh2 and Its Application
A technology of bacteria gram and lebsiella, applied in the field of environmental biology, can solve the problems of polluting the farmland in mining areas, affecting the growth of crops, reducing the quality of crops, etc. Effect
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Embodiment 1
[0015] Example 1 Isolation and screening of antimony-resistant bacteria and anti-Sb ability
[0016] Collect plant rhizosphere soil samples near the smeltery in Xikuangshan Antimony Mining Area, Lengshuijiang City, Hunan Province, weigh 100g of the above-mentioned fresh soil samples, add an appropriate amount of antimony potassium tartrate ([C 8 h 4 K 2 o 12 Sb 2 3(H 2 O)]), so that the final concentration of antimony in the soil sample was 1000mg / kg, and placed in a constant temperature incubator at 28°C for enrichment and cultivation for one week. Take 10g of the above soil sample and place it in a sterile solution containing 10 glass beads and 90mL of 0.85% NaCl, shake it in a shaker at 30°C and 180rmin-1 for 30min to fully disperse the sample. Take 0.1mL and spread it on the CDM medium (CDM medium formula: MgSO 4 ·7H 2 O 2.0 g, NH 4 Cl 1.0 g, Na 2 SO 4 1.0 g, K 2 HPO 4 0.013 g, CaCl 2 2H 2 O 0.067 g, Na-lactate 5.0 g, agar 15.0 g, add distilled water to 100...
Embodiment 2
[0019] The identification of embodiment 2 NXH2 bacterial strain
[0020]The strain was subjected to morphological, culture characteristics and sequence analysis of 16S rDNA. The 16S rDNA molecular identification was carried out according to the following steps: pick a single colony of the screened strain and inoculate it in liquid LB medium, culture it on a shaking table at 30°C and 150r / min, take out the culture medium at 24h, and centrifuge at 5000r / min for 1min to get the supernatant According to the bacterial genomic DNA extraction kit (provided by Tiangen Biochemical Technology Co., Ltd.), the colony DNA was extracted; the general primers 27F and 1492R were used for PCR amplification of the extracted bacterial DNA; the sequence of 27F was 5′-AGA GTT TGA TCC TGG CTC AG -3'; 1492R sequence is 5'-AAG GAG GTG ATC CAG CCG CA-3' primers were synthesized by Beijing Ruibo Xingke Biotechnology Co., Ltd.; the PCR products were sequenced, and the sequencing results were analyzed by ...
Embodiment 3
[0022] Example 3: Tolerance of Klebsiella NXH2 to various heavy metals
[0023] Separately prepared with different concentrations of As 3+ 、Cd 2+ 、Cr 6+ , Hg 2 CDM solid medium for heavy metals, Cd 2+ 、Cr 6+ , Hg 2 Heavy metal concentration starts from 100mg / L, followed by 100mg / L, 200mg / L, 400mg / L, 800mg / L, 1200mg / L, 1600mg / L, As 3+ Concentration starting from 100mg / L, followed by 100mg / L, 200mg / L, 400mg / L, 800mg / L, 1200mg / L, 1600mg / L, 2000mg / L, 3000 mg / L, 4000 mg / L, without adding Heavy metal treatment served as a control. Klebsiella sp. ( Klebsiella sp. ) NXH2 strain was inoculated into heavy metal-free CDM medium for activation and culture for 24 hours, and 0.1 mL was inoculated into medium containing different types and concentrations of heavy metals, and cultured at 30°C for 24 hours. , The growth situation in the culture medium of different concentrations of heavy metals, the results are shown in Table 1. It can be seen from the table that the NXH2 strain has...
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