[논문]Artificial induction and isolation of cadmium-tolerant soil bacteria

Lee, Sangman

doi:10.3839/jabc.2020.017

Artificial induction and isolation of cadmium-tolerant soil bacteria 원문보기

Journal of applied biological chemistry, v.63 no.2, 2020년, pp.125 - 129

Lee, Sangman (Division of Applied Biology and Chemistry, School of Applied Biosciences, College of Agriculture and Life Sciences, Kyungpook National University)

Abstract ▼ AI-Helper

Environmental pollution caused by various heavy metals is a serious global problem. To solve this problem, microbial bioremediation of contaminated metals has developed rapidly as an effective strategy when physical and chemical techniques are not suitable. In this study, cadmium (Cd)-tolerant soil bacteria were isolated via artificial induction in laboratory conditions instead of screening bacteria naturally adapted to metal-contaminated soils. Wild-type (WT) bacteria grown in uncontaminated soils were artificially and sequentially adapted to gradually increasing Cd concentrations of up to 15 mM. The resultant cells, named Soil-CdR15, survived at a Cd concentration of 10 mM, whereas WT cells failed to survive with 4 mM Cd on solid media for 2 d. In liquid media containing Cd, the SoilCdR15 cells grew with 15 mM Cd for 7 d, whereas the WT cells could not grow with 5 mM Cd. Both Soil-CdR15 and WT cells removed approximately 35% of Cd at the same capacity from liquid media containing either 0.5 or 1.0 mM Cd over 2 d. In addition to Cd, the Soil-CdR15 cells showed increased resistance to nickel, zinc, and arsenic compared to WT cells. The Soil-CdR cells were identified as Burkholderia sp. by partial sequencing of 16S rRNA. The data presented in this study demonstrate that isolation of heavy metal-tolerant microorganisms via artificial induction in laboratory conditions is possible and may be useful for the application of the microorganisms for the bioremediation of heavy metals.

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제안 방법

Thus, the long-exposure of microorganisms to heavy metals in artificial conditions can induce metal tolerance by inducing mutations during the adaptation process. Therefore, this study focused on the artificial generation of bacterial mutants that have a strong tolerance to Cd, a representative non-essential heavy metal, using the sequential adaptation of bacteria to gradually increases in Cd stress.

대상 데이터

A soil sample was collected from a rose garden at Kyungpook National University, Korea. Approximately 5 g soil was mixed with 10 mL Luria-Bertani (LB) medium, and 100 µL of the supernatant was inoculated in 5 mL LB medium.
8% (w/v) agarose gel electrophoresis and purified using a Gel Extraction Kit (Invitrogen, Carlsbad, CA, USA). The purified DNA fragments were sent to Solgent (Daejeon, Korea) for sequencing.
The universal primers 27F (5'-AGA GTT TGA TCC TGG CTC AG-3') and 1492R (5'-GGC TAC CTT GTT ACG ACT T-3') were used.

데이터처리

A one-way analysis of variance (ANOVA) was carried out to measure significant differences among the groups at the significant level of 5%, p <0.05 using a SPSS program with a 23.0 version.

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Artificial induction and isolation of cadmium-tolerant soil bacteria 원문보기

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Artificial induction and isolation of cadmium-tolerant soil bacteria 원문보기

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