A method for determining the content of saturated hydrocarbon degradation gene alkb in petroleum-contaminated soil

Abstract

The invention provides a method for measuring the content of a saturated hydrocarbon-degrading gene AlkB in petroleum-contaminated soil. With saturated hydrocarbon with highest content in the petroleum-contaminated soil as a target degraded component, the saturated hydrocarbon-degrading gene AlkB is quantified by adopting culture-independent molecular biological technique, namely real-time fluorescent quantitative PCR (polymerase chain reaction). The method mainly comprises the steps of extracting DNA (deoxyribonucleic acid) in the soil, designing degenerate primers for specifically amplifying the degrading gene of saturated hydrocarbon, determining PCR conditions, making a fluorescent quantitative PCR standard curve, and measuring the sample. The novel method can be used for detecting the biodegradation activity of saturated hydrocarbon, simultaneously can be used for accurately reflecting degradation and metabolism pathways of microorganisms and the petroleum contamination statuses of areas, and has important application vales in oil field contamination monitoring and control.

Description

technical field [0001] The invention belongs to the field of environmental oil pollution monitoring, in particular to a method for measuring the content of saturated hydrocarbon degradation gene AlkB in oil-polluted soil. Background technique [0002] Petroleum is an important source of energy to promote social development, but about 1×10 9 tPetroleum and its products enter groundwater, surface water and soil during petroleum exploitation, refining, storage, transportation, use and various leakage accidents, of which my country accounts for more than 600,000 tons, directly causing 5 million hectares of soil in my country to be polluted by petroleum. The saturated hydrocarbon components in petroleum components can destroy the structure of the soil, and the content is large, which has certain toxicity, and brings a series of serious problems such as reduced plant production, hidden food safety hazards, and ecological environment damage to the local area. [0003] The green, ec...

AI Technical Summary

Problems solved by technology

[0002] Petroleum is an important source of energy to promote social development, but about 1×10 9 tPetroleum and its products enter groundwater, surface water and soil during petroleum exploitation, refining, storage, transportation, use and various leakage accidents, of which my country accounts for more than 600,000 tons, directly causing 5 million hectares of soil in my country to be polluted by petroleum. The saturated hydrocarbon components in petroleum components can destroy the structure of the soil, and the content is large and has certain toxicity, which brings a series of serious problems to the local area, such as reduced plant production, hidden food safety hazards, and ecological environment damage.

[0003] The green, economical and efficient way of biodegrading petroleum has aroused the research and advocacy of many domestic scholars, but the traditional method of microbial screening and cultivation can only separate microbial species accounting for 1% to 10% of the total number of petroleum degrading bacteria, which cannot reflect the true degradation level of microorganisms at all. and reflect the diversity and distribution of microorganisms in the soil

In addition, although there have been many studies and reports on terminal monooxygenases and cytochrome P450s that encode saturated hydrocarbons, their primer design is often limited by the type of degrading bacteria, and the synthetic metabolic enzymes cannot represent most of the degrading bacteria. genotype

Images