Anti-antimony bacteria NXH3 and application thereof
A technology of bacteria and as3, applied in the field of environmental biology, can solve problems such as polluting farmland in mining areas, reducing crop quality, affecting crop growth, etc., to achieve the effects of promoting vegetation growth, alleviating the toxicity of plants to heavy metals, and broad application potential
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[0015] Example 1 Isolation and screening of anti-antimony bacteria and anti-Sb ability
[0016] Collect the plant rhizosphere soil samples near the smelter in the antimony mining area of Xikuangshan, Lengshuijiang City, Hunan Province. Weigh 100g of the above fresh soil sample and add an appropriate amount of potassium antimony tartrate ([C 8 H 4 K 2 O 12 Sb 2 ·3(H 2 O)]), make the final concentration of antimony in the soil sample 1000mg / kg, and place it in a 28°C constant temperature incubator for enrichment for one week. Take 10g of the above soil sample and place it in 10 glass beads and 90mL 0.85% NaCl sterile solution. Shake for 30 min in a shaker at 180 r min-1 at 30°C to fully disperse the sample. Take 0.1mL spread on CDM medium (CDM medium formula: MgSO 4 ·7H 2 O 2.0 g, NH 4 Cl 1.0 g, Na 2 SO 4 1.0g, 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 1000 mL, pH 7.2), in which the CDM medium uses potassium antimony tartrat...
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[0019] Example 2 Identification of NXH3 strain
[0020] The strain was analyzed by morphology, culture characteristics and 16S rDNA sequence sequencing. The 16S rDNA molecular identification is carried out according to the following steps: pick a single colony of the selected strains and inoculate it in liquid LB medium, shake culture at 30℃, 150r / min on a shaker, take out the culture medium at 24h, centrifuge at 5000r / min for 1min to take the supernatant ,According to the bacterial genomic DNA extraction kit (provided by Tiangen Biochemical Technology Co., Ltd.), extract colony DNA; universal primers 27F and 1492R are used for PCR amplification of the extracted bacterial DNA; 27F sequence is 5′-AGA GTT TGA TCC TGG CTC AG -3′; 1492R sequence is 5′-AAGGAG 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 BLAST sequenced in the NCBI database. And compare the homology.
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[0022] Example 3 Tolerance of Stenotrophomonas NXH3 to multiple heavy metals
[0023] Prepared to contain different concentrations of As 3+ , Cd 2+ , Cr 6+ , Hg 2 Heavy metal CDM solid medium, Cd 2+ , Cr 6+ , Hg 2 The concentration of heavy metals starts from 100mg / L, followed by 100mg / L, 200mg / L, 400mg / L, 800mg / L, 1200mg / L, 1600mg / L, As 3+ The concentration starts from 100mg / L, followed by 100mg / L, 200mg / L, 400mg / L, 800mg / L, 1200mg / L, 1600mg / L, 2000mg / L, 2500 mg / L, 3500 mg / L, 4500 mg / L, with no heavy metal treatment as a control. Stenotrophomonas ( Stenotrophomonas sp) NXH3 strain was inoculated into heavy metal-free CDM medium for activation and culture for 24 hours. 0.1 mL was inoculated into the medium containing different types and different concentrations of heavy metals, and incubated at 30°C for 24 hours. The results are shown in Table 1 for the growth conditions in the medium of different concentrations of heavy metals. It can be seen from the table that the NXH3 stra...
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