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Applications of Folsomia candida heat shock protein hsp70 gene in soil environment quality monitoring

A technology for environmental quality and soil, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as unclear springtails, achieve short time consumption, sensitivity at individual level, maintenance of soil safety and sustainable use Effect

Inactive Publication Date: 2013-07-24
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is unclear whether the expression of the collembola F. candida hsp70 gene can be used as a molecular marker to monitor changes in the soil environment

Method used

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  • Applications of Folsomia candida heat shock protein hsp70 gene in soil environment quality monitoring
  • Applications of Folsomia candida heat shock protein hsp70 gene in soil environment quality monitoring
  • Applications of Folsomia candida heat shock protein hsp70 gene in soil environment quality monitoring

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Experimental program
Comparison scheme
Effect test

preparation example Construction

[0056] 4. Preparation of standard curve and relative quantitative PCR

[0057] (1) Preparation of standard curve

[0058] Positive clones were randomly selected and expanded in LB liquid medium. Take about 1ml of fresh bacteria liquid for plasmid extraction. Plasmid extraction was carried out using a plasmid purification kit according to the instructions. The concentration and purity of the extracted plasmid were detected by Nanodrop, a micro-nucleic acid and protein analyzer. The purified plasmid was sequentially diluted into standard samples according to a 10-fold dilution gradient. Choose 10 in this experiment 3 -10 8 A range of standard samples was used to make a standard curve. The standard sample and the sample to be tested were subjected to fluorescent quantitative PCR, and each sample was repeated three times, and the copy number of the target gene in the sample was calculated according to the standard curve.

[0059] (2) Quantitative PCR reaction

[0060] The ...

Embodiment 1

[0065] The application of the expression of the collembola F.candida hsp70 gene in the monitoring of heavy metal Hg soil pollution is mainly divided into three steps, namely: Collembola exposure; RNA extraction and cDNA synthesis; relative quantification of the hsp70 gene. The result obtained is as figure 1 Shown, its half effect concentration EC50 value is 0.42mg / kg.

Embodiment 2

[0067] The application of the expression of the collembola F.candida hsp70 gene in the monitoring of the organophosphorus pesticide-profenofos soil pollution is mainly divided into three steps, namely: exposure of collembola; RNA extraction and cDNA synthesis; hsp70 gene relative Quantitative. The result obtained is as figure 2 As shown, its half effect concentration EC50 value is 0.23mg / kg. Among them, the minimum monitoring concentration of profenofos with statistical significance is 0.2mg / kg.

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Abstract

The invention relates to applications of Folsomia candida heat shock protein hsp70 (F. candida hsp70) gene in soil environment quality monitoring. The inventor finds that the F. candida hsp70 gene can be used for the soil environment quality monitoring. The invention provides a soil environment quality monitoring method. The method comprises the following steps: 1, exposing Folsomia candida in soil to be detected; and 2, detecting the expression level of the hsp70 gene in the exposed Folsomia candida. The method allows the soil quality to be rapidly monitored and diagnosed through treating artificially-cultured soil animal Folsomia candida as an experiment and using the F. candida hsp70 gene expression level within 48h as a soil quality monitoring index. The method can be used for the early stage diagnosis and early warning polluted soil, and the soil quality assessment, and is more sensitive and rapid than traditional toxicity test methods.

Description

technical field [0001] The invention relates to a method for using environmental biomarkers to monitor soil pollution. Specifically, the present invention relates to the use of the dose-effect relationship between the hsp70 gene expression of quantitative collembola (Folsomia candida) and the soil pollution level to characterize the toxicity of polluted soil, which is suitable for the diagnosis, environmental early warning and ecological risk of soil environmental pollution Evaluate. Background technique [0002] Soil pollution is an increasingly serious environmental problem, which directly threatens the safety of human food health and the sustainable development of soil ecological functions. Toxicity and risk assessment of contaminated soils have attracted widespread attention, but only a combination of chemical and biological methods can effectively detect the interaction between soil pollutants and organisms. [0003] As an important model organism, F. candida is often...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 贺纪正刘玉荣郑袁明
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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