CN210381991U

CN210381991U - Test device for exploring in-situ rhizosphere PAHs degradation functional microorganisms

Info

Publication number: CN210381991U
Application number: CN201920823229.3U
Authority: CN
Inventors: 李继兵; 罗春玲; 李军; 张干
Original assignee: Guangzhou Institute of Geochemistry of CAS
Current assignee: Guangzhou Institute of Geochemistry of CAS
Priority date: 2019-05-31
Filing date: 2019-05-31
Publication date: 2020-04-24
Anticipated expiration: 2029-05-31

Abstract

The utility model discloses a probe test device of normal position rhizosphere PAHs degradation function microorganism, including planting the box, be equipped with the contaminated soil who contains PAHs in planting the box, it has the plant to plant on the contaminated soil, still includes the glass pipe, and the intraductal mark soil that contains stable isotope mark that is equipped with of glass, the glass pipe is buried underground in contaminated soil, and the partial root system of plant stretches into in the mark soil of glass pipe. The utility model discloses a test device introduces rhizosphere normal position research with SIP, helps understanding which microorganism has participated in the pollutant degradation in the actual rhizosphere environment, what its degradation mechanism is, and how the interrelation between the degradation function microorganism also can probe into the influence mechanism of plant rhizosphere effect to function microorganism kind and metabolic capability simultaneously, provides more theoretical foundation for microbial remediation and rhizosphere repair technique.

Description

Test device for exploring in-situ rhizosphere PAHs degradation functional microorganisms

Technical Field

The utility model relates to a probe test device of normal position rhizosphere PAHs degradation function microorganism.

Background

Polycyclic Aromatic Hydrocarbons (PAHs) are a class of hydrocarbons containing two or more fused benzene ring structures in the molecule, and are representative of Persistent Organic Pollutants (POPs) (Edwards, 1983). Such substances can enter the body through the skin, respiratory tract and digestive tract, constituting a significant threat to the ecological environment and human health (Grova et al, 2011; Li et al, 2017; Xiong et al, 2017). In view of their adverse effects on human health and ecological environment, PAHs are the focus of attention of the government and academia of various countries (Meng et al, 2019). Therefore, an efficient and cheap PAHs pollution remediation method is sought, the elimination speed of the PAHs in the environment is improved, the degradation process of the PAHs in the environment is proved, the content of the PAHs in the environment is kept at a lower concentration level, and the method has important practical significance for preventing and treating soil pollution, guaranteeing human health and ecological safety and realizing economic and environmental sustainable development. Microbial degradation is one of the main ways for eliminating PAHs in soil under natural conditions, and is also a main remediation measure for PAHs contaminated soil. The traditional separation culture technology is adopted to separate and obtain the high-efficiency PAHs degrading bacteria from the polluted environment, which is a key step of microbial remediation (Bartha, 1986; Wei et al, 2017; Li et al, 2018 a). However, this method has great limitations. Only 0.1-1% of culturable microorganisms can be obtained in the laboratory at present; a large number of environmental factors, such as osmotic pressure, temperature, and other environmental conditions, make the in situ environment significantly different from pure culture (Schut et al, 1993; Varela et al, 2004); factors such as alteration of niches under traditional culture conditions, disruption of co-metabolism and quorum sensing systems also lead to reduced culturability of the microorganism (Guan et al, 2000; Bruns et al, 2002). Therefore, the research on the whole functional microbial community (including culturable and non-culturable microorganisms) actually participating in the PAHs degradation in the in-situ environment and the mutual relationship thereof has important significance for breaking through the bottleneck of the traditional PAHs degradation microorganism research.

Stable Isotope Probe (SIP) technology is a technology that combines the structure and function of microbial communities in complex environments. The technology can directly detect which microorganisms in the environment participate in the degradation of PAHs without carrying out isolated culture on the microorganisms, and becomes an important means for identifying functional microorganisms in situ (Song et al, 2015; Jiang et al, 2018; Li et al, 2018 b). The technology is used for determining a functional microbial community in a sample by adding a proper amount of a stable isotope labeled substrate to the sample, separating and analyzing an isotope labeled biomarker after finishing culture and combining a molecular biology method (Dumont and Murrell, 2005). Stable isotopes currently used for labelling include¹³C、¹⁵N、¹⁸O, and the like. The basic operation flow of the technology mainly comprises the following steps: firstly, in-situ or microcosm environmental samples are exposed to a pollutant matrix containing stable isotope labels for culture; then the microorganisms capable of metabolizing by utilizing the isotope labeled substrate in the sample can be subjected to assimilation synthesis to form the biomarker containing the isotope label; and finally, collecting the sample, extracting, separating, purifying, measuring and analyzing the biomarker, and identifying the microorganisms with specific functions in the environmental sample. At present, SIP technology is applied to explore PAHs degradation functional microorganisms in environments such as soil, sewage and the like, and a plurality of functional microorganisms with in-situ degradation are identified.

The rhizosphere microorganisms are actively metabolized, are the areas where PAHs are reduced most rapidly, and have special positions in the degradation process of the PAHs in the nature. The rhizosphere is a unique circle formed by the interaction of a plant root system and soil microorganisms, and is a place for the interaction of the plant-soil-microorganisms and the environment. The abnormally developed root system of the plant provides a habitat and a suitable growing environment for microorganisms. The plant rhizosphere can increase the air permeability and permeability of soil and the dispersion of soil aggregates, thereby improving the metabolic activity and diversity of rhizosphere microorganisms and simultaneously increasing the bioavailability of PAHs (Hamdi et al, 2007). In addition, the root system of the plant can secrete a large amount of carbohydrates such as amino acids and organic acids, so that nutrition is provided for the growth and the propagation of microorganisms, the content of the carbohydrates reaches dozens or even thousands of times of that of non-rhizosphere, and some secretion components can be used as surfactants to increase the solubility of PAHs (Cebron et al, 2011). Rhizosphere soil has lower organic pollutant content, higher degradation rate, and is often accompanied by an increase in the diversity and number of degrading microorganisms, as compared to non-rhizosphere soil (Cebron et al, 2011; huangjunwei et al, 2017). Plant rhizosphere environment provides convenient conditions for organic pollutant remediation, and is being widely valued by researchers in various countries.

Currently, SIP research on PAHs degradation is limited to microcosm experiments, but no research is conducted on in-situ rhizosphere environment. Therefore, it is necessary to design a test device capable of exploring the in situ degradation function of PAHs to solve the practical problem.

SUMMERY OF THE UTILITY MODEL

To the problem that exists among the above-mentioned prior art, the utility model provides a probe test device of normal position rhizosphere PAHs degradation function microorganism can realize the research to normal position rhizosphere PAHs degradation function microorganism.

In order to achieve the above purpose, the technical scheme of the utility model is that:

the utility model provides a probe test device of normal position rhizosphere PAHs degradation function microorganism, includes plants the box, is equipped with the contaminated soil that contains PAHs in planting the box, has planted the plant on the contaminated soil, still includes the glass pipe, is equipped with the mark soil that contains stable isotope mark in the glass pipe, and the glass pipe is buried underground in contaminated soil, and the partial root system of plant stretches into in the mark soil of glass pipe.

As an improvement of the utility model, the planting box be detachable rectangle box body, including roof, curb plate and bottom plate, the roof is equipped with the through-hole that supplies the plant to pass, and the curb plate is transparent glass board, and the bottom plate is surface covering glass fiber's perforated plate.

Compared with the prior art, the utility model, its beneficial effect lies in:

the utility model discloses a test device introduces rhizosphere normal position research with SIP, helps understanding which microorganism has participated in the pollutant degradation in the actual rhizosphere environment, what its degradation mechanism is, and how the interrelation between the degradation function microorganism also can probe into the influence mechanism of plant rhizosphere effect to function microorganism kind and metabolic capability simultaneously, provides more theoretical foundation for microbial remediation and rhizosphere repair technique.

Drawings

FIG. 1 is a schematic structural view of a test apparatus of the present invention;

description of reference numerals: 1-a bottom plate; 2-side plate; 3-a top plate; 4-contaminated soil; 5-a plant; 6-root system; 7-a glass tube; 8-bolt assembly.

Detailed Description

In order to make the above objects, features and advantages of the present invention more comprehensible, the present invention is described in detail with reference to the accompanying drawings and the detailed description.

As shown in figure 1, the test device for researching in-situ rhizosphere PAHs degradation functional microorganisms comprises a planting box and a glass tube 7. The planting box is filled with polluted soil 4 containing PAHs, and plants 5 are planted on the polluted soil 4. The planting box is a detachable rectangular box body and comprises a bottom plate 1, four side plates 2 and a top plate 3, wherein the bottom plate 1 is a porous plate with the surface covered with glass fiber, and the planting box has the functions of improving the drainage effect and the air permeability of a soil medium in the box and avoiding the root system of a plant from mildewing and rotting due to too much watering or blocked breathing; the side plates 2 are transparent glass plates, the four side plates 2 are detachably arranged between the bottom plate 1 and the top plate 3 through bolt assemblies 8, and the side plates 3 needing to be opened can be selected according to the growth condition of a plant root system in the using process; the top plate is provided with a through hole for the plant 5 to pass through, and the through hole is also used for watering.

The glass tube 7 is filled with labeled soil containing stable isotope label which can be selected¹²C-PAHs and¹³C-PAHs marks, the glass tubes 7 are buried in the polluted soil, and part of root systems 6 of the plants 5 extend into the marked soil of the glass tubes 7, so that the combination of SIP and in-situ rhizosphere is realized.

During the use, the planting box is adorned and is polluted soil well, plants and waits to carry out normal position rhizosphere SIP experiment after the vegetation reaches certain time (because of the plant difference), selects the glass curb plate that needs to open according to the growth condition of plant roots, opens the curb plate after, introduces the root system respectively to being equipped with and is equipped with¹²C-PAHs and¹³after the glass tube of the soil marked by the C-PAHs is cultured for a period of time, the glass tube with different treatments can be taken out, and the soil in the glass tube is directly analyzed by biomarkers, metabolic intermediates and degradation genes.

The above embodiments are only for illustrating the technical concept and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the contents of the present invention and to implement the present invention, which cannot limit the protection scope of the present invention. All equivalent changes and modifications made according to the spirit of the present invention should be covered by the protection scope of the present invention.

Claims

1. A test device for exploring in-situ rhizosphere PAHs degradation functional microorganisms is characterized in that: the soil-planting device comprises a planting box, wherein the planting box is internally provided with contaminated soil containing PAHs, plants are planted on the contaminated soil, the glass tube is internally provided with labeled soil containing stable isotopes, the glass tube is buried in the contaminated soil, and partial root systems of the plants extend into the labeled soil of the glass tube.

2. The device of claim 1, wherein the device comprises: the planting box be detachable rectangle box body, including roof, curb plate and bottom plate, the roof is equipped with the through-hole that supplies the plant to pass, the curb plate is transparent glass board, the bottom plate is the porous plate of surface covering glass fiber.