CN114134020A

CN114134020A - Nondestructive sampling device and nondestructive sampling method for plant rhizosphere microorganisms

Info

Publication number: CN114134020A
Application number: CN202111453232.9A
Authority: CN
Inventors: 谢剑波; 刘思佳; 陈思思; 谭书贤
Original assignee: Beijing Forestry University
Current assignee: Beijing Forestry University
Priority date: 2021-12-01
Filing date: 2021-12-01
Publication date: 2022-03-04

Abstract

The invention belongs to the technical field of experimental sampling, and particularly relates to a nondestructive sampling device and a nondestructive sampling method for plant rhizosphere microorganisms. The invention provides a nondestructive sampling device for plant rhizosphere microorganisms, which comprises an inner layer mesh container and at least one outer layer mesh container. The nondestructive sampling device provided by the invention can not only ensure that plant materials are prevented from being damaged, but also ensure that rhizosphere microorganisms of the same plant are not changed due to root system damage, and further can sample the same plant at multiple time points; and can the ration sample, the sample is convenient quick more. Therefore, the device provided by the invention can provide technical support for researches such as dynamic change and regulation network of plant rhizosphere microorganisms.

Description

Nondestructive sampling device and nondestructive sampling method for plant rhizosphere microorganisms

Technical Field

The invention belongs to the technical field of experimental sampling, and particularly relates to a nondestructive sampling device and a nondestructive sampling method for plant rhizosphere microorganisms.

Background

The rhizosphere is the area where the life activities and metabolism of the root system have the most direct and most intense influence on the soil, is an important place where soil microorganisms act on plants, and is specially a thin layer of soil around the root system of the plants. Rhizosphere microorganisms are specific microorganism flora living in plant rhizosphere and directly influenced by root systems, and have profound influences on various aspects such as plant growth, nutrition and health (Wangxiaoling and the like, a mechanism for promoting the utilization of rice nitrogen by rhizosphere microorganisms [ J ]. plant science and newspaper, 2019,54(03): 285-. With the progress of research, research on rhizosphere microorganisms is increasingly emphasized, and the key functions of root microorganisms are to improve nutrient absorption, promote plant growth and development, improve root system structure, and protect hosts from biotic and abiotic stresses (wangma et al. Weak waxberry rhizosphere soil microorganism diversity research [ J ] Zhejiang agricultural science, 2021,62(06): 1128+ 1132.).

Although there are many studies on rhizosphere microorganisms at home and abroad, the sampling mode is single. Typically, plant rootlets are collected, followed by removal of soil that does not adhere tightly to the roots and washing with PBS buffer (Xu Jin et al. the structure and function of the biological ecology microbiome [ J ]. Nature communications,2018,9(1): 4894; Xu Ling et al. Dry delay degradation of the sorghum microorganisms and enzymes for monoderm bacteria [ J ]. Proceedings of the National Academy of science of the United States of America,2018,115(18): E4284-E4293). The sampling process is complicated, the root of the whole plant is almost used for one-time sampling, and the plant is difficult to survive after sampling. In order to complete the corresponding experiment, a large number of plants are usually consumed, which not only causes a large amount of waste of experimental materials, but also increases errors undoubtedly for the experiment needing to observe the dynamic change of rhizosphere microorganisms in a combined sampling mode of a plurality of plants, and increases greater difficulty for the deeper research of the rhizosphere microorganisms.

Disclosure of Invention

In order to overcome the technical problems, the invention provides a nondestructive sampling device and a nondestructive sampling method for plant rhizosphere microorganisms.

In order to achieve the above purpose, the invention provides the following technical scheme:

the invention provides a nondestructive sampling device for plant rhizosphere microorganisms, which comprises an inner layer mesh container and at least one outer layer mesh container.

Preferably, the bottom of the inner-layer mesh container is provided with a cross opening.

Preferably, the mesh aperture of the inner layer mesh container is more than or equal to 30 μm; the mesh aperture of the outer layer mesh container is 50-150 mu m.

The method for nondestructively sampling the rhizosphere microorganisms of the plants by adopting the nondestructive sampling device in the technical scheme comprises the following steps: and moving the seedlings with soil into an inner-layer reticular container of the nondestructive sampling device, wherein the periphery of the inner-layer reticular container surrounds at least one outer-layer reticular container, placing the nondestructive sampling device in the soil, continuously culturing until the roots of the plants spread the whole inner-layer reticular container, and then taking at least one outer-layer reticular container for sampling.

Preferably, the plant comprises poplar, tomato, eggplant or rice.

Preferably, the seedling is a robust seedling; the culture method of the seedling with strong growth comprises the steps of dipping the sterile tissue culture seedling into a hormone sterilization solution for root treatment, and carrying out sterile culture to obtain the seedling with strong growth.

Preferably, the hormone sterilization solution comprises a hormone and a sterilant; the hormone comprises indolebutyric acid or 1-naphthylacetic acid; the sterilizing agent comprises carbendazim or zineb.

Preferably, the concentration of the hormone in the hormone sterilization solution is 0.0075-0.015 mg/mL, and the concentration of the sterilizing agent is 0.5-1% by mass.

Preferably, the time for root dipping treatment is 10s to 60 s.

Preferably, the characteristics of robust growth include large and spread leaves, bright green color and strong and straight and tough essence.

Has the advantages that:

the invention provides a nondestructive sampling device for plant rhizosphere microorganisms, which comprises an inner layer mesh container and at least one outer layer mesh container. When the sampling device is used, the periphery of the inner-layer mesh container is surrounded by at least one outer-layer mesh container, the nondestructive sampling device provided by the invention is used for sampling plant rhizosphere microorganisms, the mode of collecting plant fine roots to obtain rhizosphere soil is not needed, the plant rhizosphere soil can be obtained only by extracting the outer-layer mesh container, and then rhizosphere microorganism samples are obtained, so that plant materials can be prevented from being damaged due to the collection of the plant fine roots, the rhizosphere microorganisms of the same plant cannot be changed due to the damage of the root system, the sampling of the same plant at different time points can be realized, and the multi-time-point sampling of the same plant can be realized.

Simultaneously, can select to set up an outer netted container or a plurality of outer netted containers according to the needs of experiment sample, and then reach the quantitative sampling of plant rhizosphere microorganism, the sample is convenient more quick. Therefore, the device provided by the invention can provide technical support for researches such as dynamic change and regulation network of plant rhizosphere microorganisms.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments will be briefly described below.

FIG. 1-A is a schematic view of a non-destructive sampling apparatus;

FIG. 1-B is a schematic view of the structure of the inner mesh container;

FIG. 1-C is a schematic view of the structure of an outer layer mesh container;

FIG. 2-A is a perspective view of an example lossless sampling apparatus;

FIG. 2-B is a top view of an example non-destructive sampling device;

FIG. 2-C is a front view of an example non-destructive sampling device;

FIG. 3 is a graph showing the results of the microbial composition of soil samples collected in example 1;

wherein 1 is the netted container of inlayer, 2 is the netted container of outer, and 3 are no end hollow structure, and 4 are the rope on the netted container of inlayer, and 5 are the rope on the netted container of outer.

Detailed Description

In the invention, the nondestructive sampling device sequentially comprises an inner-layer mesh container and at least one outer-layer mesh container from inside to outside. In use, the inner mesh container will surround at least one outer mesh container at the periphery of the inner mesh container. As shown in FIG. 1-A, the present invention provides a nondestructive sampling device comprising an inner mesh container 1, and at least one outer mesh container 2; the outer layer net structure is located the outside of inlayer net structure, and when having a plurality of outer layer net structure, it is a plurality of outer layer net structure with inlayer net structure peripheral line places in order, forms the parcel inlayer net structure's inner chamber.

When used, the non-destructive sampling device preferably further comprises a bottomless hollow structure 3. In the invention, the bottomless hollow structure is positioned outside the outer-layer reticular container, and when a plurality of outer-layer reticular structures are arranged, the outer-layer reticular structures are sequentially arranged on the inner periphery of the bottomless hollow structure and can wrap the inner-layer reticular container.

In the present invention, the bottomless hollow structure has an effect of assisting in fixing the inner-layer mesh container and the outer-layer mesh container. In the present invention, the shape of the bottomless hollow structure preferably conforms to the shape of the inner mesh container, and more preferably is a cylinder. The material of the bottomless hollow structure is not specially limited, and any material capable of supporting the shape of the bottomless hollow structure can be adopted. The size of the bottomless hollow structure is not particularly limited in the invention, and the bottomless hollow structure can be wrapped and fixed on the inner-layer reticular container and the outer-layer reticular container, such as a hollow cylinder with the bottom diameter of 8cm and the height of 20cm is adopted in the embodiment of the invention.

According to the invention, after the nondestructive sampling device is assembled, the bottomless hollow structure is preferably removed.

In the present invention, the mesh diameter of the inner mesh container is preferably not less than 30 μm, more preferably 30 to 50 μm, and still more preferably 50 μm. The aperture can meet the requirement that microorganisms such as bacteria, fungi and the like can freely pass through; the upper limit of the pore diameter of the internal reticular structure is not specially limited, and the plant can only grow out after the roots of the plant are spread.

The size of the internal reticular container is not specially limited, and the sampling requirement of the corresponding plant can be met. When the plant is poplar, the inner-layer reticular container is preferably a cuboid and a cylinder; the diameter of the bottom of the cylinder is preferably 6-10 cm, and more preferably 6 cm; the height of the cylinder is preferably 18-25 cm, and more preferably 18 cm. The material and the structure of the inner-layer reticular container are not specially limited, and the requirement of sampling corresponding plants can be met, for example, a nylon mesh bag is specifically adopted in the embodiment of the invention.

In the invention, the inner-layer netted container is preferably a cover-free container, the bottom of the inner-layer netted container is preferably provided with cross openings, and the ratio of the diameter and the number of the cross openings to the inner diameter of the inner-layer netted container is preferably 0.8-1.2 cm: 2-4: 6 cm; the cross openings can ensure that the water can permeate and the root system can extend out after fully spreading, for example, in the embodiment of the invention, a cylindrical inner-layer reticular container with the bottom diameter of 6cm is preferably adopted for sampling the rhizosphere microorganisms of the poplar, and the circular bottom of the inner-layer reticular container preferably comprises two cross openings of 0.8-1.2 cm, and more preferably comprises two cross openings of 1 cm. The cross openings are distributed on two sides of the central point on the same diameter of the bottom.

As shown in fig. 1-B, the inner mesh container 1 preferably further includes a string 4 thereon. The rope is preferably arranged at the opening of the inner-layer reticular container, the existence mode and the material of the rope are not particularly limited, and the nondestructive sampling device is only convenient to assemble. In the present invention, the number of the outer layer mesh containers is at least 1, and more preferably, the peripheral mesh containers are required to be arranged in a circle along the periphery of the inner layer mesh container.

In the invention, the mesh aperture of the outer layer mesh container is preferably 50-150 μm, more preferably 100-150 μm, and further preferably 150 μm; the aperture ensures that the soil is not leaked out.

In the present invention, the height of the outer mesh container is preferably kept consistent with the height of the inner mesh container; the height ensures consistency of rhizosphere microorganisms of the inner layer mesh container and the outer layer mesh container. The invention has no special limitation on other sizes of the outer-layer reticular container, and the sampling quantity of a single outer-layer reticular container corresponding to the plant rhizosphere sample can be met. The material and the structure of the outer layer reticular container are not specially limited. When the plant is poplar, the outer-layer mesh container is preferably rectangular; the height of the rectangular outer-layer mesh container can be preferably 18-25 cm, and more preferably 18 cm; the width is preferably 1.5-2 cm, and more preferably 2 cm. In the embodiment of the invention, a rectangular nylon mesh bag with the height of 18cm, the width of 2cm and the aperture of 150 mu m is specifically adopted.

As shown in fig. 1-C, the outer mesh container 2 preferably further comprises a string 5 thereon. The rope is arranged at the opening of the outer layer mesh container, so that the sampling is convenient. The invention has no special limit on the existence mode and the material of the rope, and only needs to meet the requirement that the outer layer reticular container is conveniently pulled out by the rope during sampling.

In the present invention, the method for constructing the lossless sampling apparatus preferably includes the steps of:

after inserting the bottomless hollow structure into soil, discharging the outer-layer netted container in the bottomless hollow structure to form a bottomless hollow structure comprising the outer-layer netted container;

placing the inner-layer reticular container in the bottomless hollow structure comprising the outer-layer reticular container to obtain the nondestructive sampling device;

the outer layer netted container in the soilless hollow structure is positioned at the periphery of the inner layer netted container.

The invention preferably inserts the bottomless hollow structure into the soil to form a soilless hollow structure placed in the soil. The soil is preferably present in the planting device or the field, more preferably in the planting device. In the present invention, the soil is preferably a soil having bacteria.

After the soilless hollow structure placed in the soil is obtained, the outer-layer netted container is preferably arranged inside the soilless hollow structure to form the soilless hollow structure comprising the outer-layer netted container. In the invention, the outer layer of reticular containers are preferably arranged along the inner wall of the soilless hollow structure for a circle; the outer layer reticular container is preferably clung to the inner wall of the soilless hollow structure.

After the soilless hollow structure comprising the outer-layer netted container is obtained, the inner-layer netted container is preferably placed in the soilless hollow structure comprising the outer-layer netted container, so that the nondestructive sampling device is obtained; the outer layer netted container in the soilless hollow structure is preferably positioned at the periphery of the inner layer netted container.

In practice, the invention also preferably comprises withdrawing the soil-less hollow structure from the non-destructive sampling device. In the invention, the soilless hollow structure has the function of assisting in fixing and forming the nondestructive sampling device, and more particularly, the function of facilitating the placement of the inner reticular container and the outer reticular container in the nondestructive sampling device in soil.

The invention also provides a method for nondestructively sampling plant rhizosphere microorganisms by adopting the nondestructive sampling device, which comprises the following steps: transferring the seedlings with soil to an inner-layer reticular container of the nondestructive sampling device, continuously culturing until plant roots spread the whole inner-layer reticular container, and sampling the outer-layer reticular container; the nondestructive sampling device is placed in soil.

In the present invention, the plant preferably includes poplar, tomato, eggplant or rice, more preferably includes poplar and tomato, and further preferably includes poplar.

In the present invention, the seedling preferably includes a seedling which is strongly grown; the culture method of the strong seedling preferably comprises the steps of dipping the sterile tissue culture seedling into a hormone sterilization solution for root treatment, and performing sterile culture to obtain the strong seedling.

The invention preferably treats the aseptic tissue culture seedling with the hormone sterilized solution to dip the root to obtain the aseptic tissue culture seedling treated with the root.

The invention has no special limitation on the obtaining of the sterile tissue culture seedling, and the conventional sterile tissue culture technology in the field is adopted. When the plant is a poplar, the medium of the sterile tissue culture seedling preferably consists of the following components in concentration: 2.18 g/L1/2 MS, 0.05-0.1 mg/L PNA, 0.05-0.1 mg/L IBA, 18-23 g/L sucrose and 5.5-7 g/L agar, more preferably 2.18 g/L1/2 MS, 0.1mg/L LNAA, 0.1mg/L IBA, 20g/L sucrose and 6g/L agar. The growth state of the sterile tissue culture seedling is not particularly limited, and the sterile tissue culture seedling can reach the conventional transplantable state in the technical field, for example, when the plant is a poplar, the tissue culture seedling growing about 8cm of the poplar is preferably selected; the 8cm poplar tissue culture seedlings grow to be full of tissue culture bottles, plants stand upright, and leaves stretch to be emerald green.

In the present invention, the hormone sterilization solution preferably comprises a hormone and a sterilizing agent; the hormone preferably comprises indolebutyric acid (IBA) or 1-naphthaleneacetic acid (NAA), more preferably indolebutyric acid (IBA); the sterilizing agent preferably comprises carbendazim or zineb, more preferably comprises carbendazim. In the invention, the concentration of the hormone in the hormone sterilization solution is preferably 0.005-0.015 mg/L, and more preferably 0.01 mg/L; the concentration of the carbendazim is preferably 0.75-1.25%, and more preferably 1%; the dosage of the hormone sterilization solution is not specially limited, and the requirement of the conventional root dipping means in the field on root dipping of corresponding plants can be met. For example, when the plant is poplar, 200mL of the hormone sterilizing solution containing 0.01mg/L of indolebutyric acid and 1% of carbendazim is preferably adopted for root dipping treatment of poplar in the invention. In the invention, the time for root dipping treatment is preferably 10-60 s, and more preferably 10 s.

The invention also preferably carries out sterile culture on the obtained sterile tissue culture seedlings dipped with roots to obtain robust seedlings.

The sterile culture method is not particularly limited, and the conventional sterile culture method in the field can be adopted. When the plant is a poplar, the invention preferably transplants the sterile tissue culture seedling dipped with the root into a plug tray filled with sterilized soil for culture; the plug is preferably a plug with 21 holes and the height of the plug is 15 cm. The used sterilized soil preferably comprises humus soil and vermiculite, and the volume ratio of the humus soil to the vermiculite is preferably (0.8-1.2): (0.8-1.2), and more preferably 1: 1. The soil sterilization mode and sterilization equipment are not particularly limited, and the soil sterilization mode can be realized by adopting a conventional sterilization mode in the field, such as sterilization at 121 ℃ for 20min in an autoclave.

In the present invention, the growth characteristics of the well-growing seedling are preferably leafy and extended, bright green in color and tough and straight. In the invention, the characteristic of robust growth can also be preferably evaluated by adopting the height or growth time of seedlings, for example, when the sterile tissue culture seedlings dipped with roots are transplanted into sterilized soil to grow for 30-45 days and the height is 15-20 cm, the seedlings can also be determined to be robust. The invention has no special limit on the days and the height of the growth of the sterilized tissue culture seedlings dipped with the roots in the sterilized soil, and can continue to grow and spread roots in the following process according to the growth characteristics of plants.

After the seedlings with strong growth are obtained, the obtained seedlings with strong growth are preferably moved into the inner-layer reticular container of the nondestructive sampling device with soil, and are cultured continuously until the whole inner-layer reticular container of the plant root vines is obtained, and then the outer-layer reticular container is taken for sampling. In the invention, the soil-carrying transplanting mode can ensure the integrity and the indestructibility of the root system. In the present invention, the outer layer mesh container is preferably filled with the bacteria-containing soil before the strong seedlings are transplanted, and the outer layer mesh container is more preferably filled with the bacteria-containing soil.

In the present invention, the non-destructive sampling device is placed in soil, preferably, the soil is a soil with bacteria. After the seedlings with strong growth are moved to the internal netted container of the nondestructive sampling device, the method also preferably comprises the step of covering a layer of soil on the nondestructive sampling device, wherein the soil is preferably the soil with bacteria; the thickness of the soil is not particularly limited by the invention, and the opening of the nondestructive sampling device is ensured to be completely covered by the soil, such as 5cm in the sampling of the poplar rhizosphere microorganisms in the embodiment of the invention. The source of the bacteria-bearing soil is not particularly limited, and the conventional bacteria-bearing soil suitable for plant growth in the field can be adopted.

The invention has no special limit on the state of the plant root spreading the whole inner layer reticular container, and can sample according to the time point of experimental design after the root spreading the whole inner layer reticular container. In the present invention, the sampling of the outer mesh container preferably includes taking out one or more outer mesh containers, and more preferably includes pulling out the outer mesh container through a rope at an opening of the outer mesh container to obtain a rhizosphere microorganism sample.

In order to further illustrate the present invention, the following detailed description of the technical solutions provided by the present invention is made with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.

Example 1

Taking 50 poplar tissue culture seedlings in a sterile rooting culture medium, removing the culture medium attached to the roots at medium temperature in clear water, immersing the roots in 200ml of mixed solution (0.01mg/L IBA and 1% carbendazim) for 10s, and transferring the roots to a plug tray filled with sterilized soil (the ratio of humus to vermiculite is 1: 1).

And when the seedlings of the poplar are transferred to sterilized soil for 30 days, the seedlings of the poplar grow to about 15cm, at the moment, the whole seedlings of the poplar in the plug are taken out with soil, and the seedlings are placed into a cylindrical nylon mesh bag with the aperture of 50 mu m, the diameter of the bottom of the cylindrical nylon mesh bag being 6cm, the height of the cylindrical nylon mesh bag being 18cm, and two cross openings with the diameter of 1cm are arranged near the outer side of the bottom of the cylindrical nylon mesh bag.

Inserting a bottomless hollow cylinder with the bottom diameter of 8cm and the height of 20cm into a planting groove filled with the fungus soil, digging out the soil in the cylinder, vertically placing 8 rectangular nylon mesh bags with the pore diameter of 150 mu m, the length of 18cm and the width of 2cm, filled with the fungus soil, in the outermost side in the cylinder, placing the cylindrical nylon mesh bags filled with poplar seedlings in the center of the cylinder, and finally covering a layer of fungus soil with the thickness of about 5cm on the whole device to ensure that the mouths of the nylon mesh bags are completely covered by soil.

The device is successfully arranged, and the nondestructive sampling of plants can be realized.

After the device is arranged for one week, most plants survive (47 plants), and the rest plants are healthy, and the leaves have no scab. After the poplar seedlings grow stably for 45 days, the average plant height of the plants is about 25cm, the plants are strong, and the next experiment can be carried out. FIG. 2-A is a panoramic view of the nondestructive device planted with young poplar seedlings; FIG. 2-B is a top view of the nondestructive device planted with poplar seedlings; FIG. 2-C is a front view of the nondestructive device planted with poplar seedlings.

And directly extracting the rectangular nylon mesh bag for sampling according to the experimental design time point. After sequencing of the amplicon and analysis of 16S rDNA, the results are shown in table 1 and fig. 3, and it can be seen that the soil sample obtained in example 1 includes 22 phyla, 673 microorganisms of 105 genera, and the species composition is abundant, and the results are successful.

Table 1 example 1 results of composition of microorganisms in soil samples

Example 2

Taking 50 poplar tissue culture seedlings in a sterile rooting culture medium, removing the culture medium attached to the roots at medium temperature in clear water, and transferring the seedlings to a plug tray filled with sterilized soil (the ratio of humus soil to vermiculite is 1: 1).

And when the seedlings are transferred to the sterilized soil for 30 days, the seedlings of the poplar grow to about 15cm, at the moment, the whole seedlings of the poplar in the hole tray are taken out with soil, and the seedlings are placed into a cylindrical nylon mesh bag with the aperture of 50 mu m, the bottom diameter of 6cm and the height of 18 cm.

Inserting a bottomless hollow cylinder with the bottom diameter of 8cm and the height of 20cm into a planting groove filled with the fungus soil, digging out the soil in the cylinder, vertically placing 8 rectangular nylon mesh bags with the pore diameter of 150 mu m, the length of 18cm and the width of 2cm filled with the fungus soil on the outermost side in the cylinder, and placing the cylindrical nylon mesh bags filled with poplar seedlings in the center of the cylinder.

However, after the device is arranged for one week, the seedling is in a poor state, leaves and plants are wilted and die in large quantities, and when the central nylon mesh bag is taken out, the situation that the bottom of the cylindrical nylon mesh bag is not provided with a cross structure and is not easy to drain water is found, so that the plant is excessively moisturized, and a rhizosphere sample of a good-growing plant cannot be obtained.

Example 3

When the seedlings of the poplar are moved to sterilized soil for 30 days, the seedlings of the poplar grow to about 15cm, at the moment, the whole seedlings of the poplar in the hole tray are taken out with soil, and are placed into a cylindrical nylon mesh bag with the hole diameter of 50 mu m, the bottom diameter of 6cm, the height of 18cm and two cross openings with the diameter of 1cm close to the outer side of the bottom, and the plants are placed in the nylon mesh bag through the cross openings so as to be convenient for water to flow.

After one week of plant placement, a small number of plants died (8 plants), but the remaining plant leaves had lesions. After the plants grow for one month, the average plant height of the seedlings is 10cm, the state is general, and rectangular nylon mesh bags are directly extracted according to the experimental design time points for sampling.

After the experiment result is subjected to amplicon sequencing and 16SrDNA analysis, the species composition is single, and the experiment result is poor.

Comparative example 1

The pore size of the inner mesh container was 150 μm compared to example 1.

Root system in the cylinder nylon pocket can stretch out the pocket from each position, even twine in the outer nylon pocket, can't realize harmless sample, and then can't satisfy the experiment demand.

From the above embodiments, it can be known that the nondestructive sampling device provided by the present invention can be used for quantitatively sampling rhizosphere microorganism samples, can realize multi-time sampling of the same plant sample, and can ensure the quality of the sample and the avoidance of plant damage.

Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.

Claims

1. A non-destructive sampling device for plant rhizosphere microorganisms is characterized by comprising an inner layer mesh container and at least one outer layer mesh container.

2. The non-destructive sampling device of claim 1, wherein the bottom of said inner mesh container is provided with a cross opening.

3. The non-destructive sampling device of claim 1, wherein said inner mesh container has a mesh size of 30 μm or more; the mesh aperture of the outer layer mesh container is 50-150 mu m.

4. A method for non-destructive sampling of plant rhizosphere microorganisms using the non-destructive sampling device of any one of claims 1 to 3, comprising the steps of:

and moving the seedlings with soil into an inner-layer reticular container of the nondestructive sampling device, wherein the periphery of the inner-layer reticular container surrounds at least one outer-layer reticular container, placing the nondestructive sampling device in the soil, continuously culturing until the roots of the plants spread the whole inner-layer reticular container, and then taking at least one outer-layer reticular container for sampling.

5. The method of claim 4, wherein the plant comprises poplar, tomato, eggplant or rice.

6. The method of claim 4 or 5, wherein the seedling is a robust growing seedling; the culture method of the seedling with strong growth comprises the steps of dipping the sterile tissue culture seedling into a hormone sterilization solution for root treatment, and carrying out sterile culture to obtain the seedling with strong growth.

7. The method of claim 6, wherein the hormone sterilization solution comprises a hormone and a sterilant;

the hormone comprises indolebutyric acid or 1-naphthylacetic acid; the sterilizing agent comprises carbendazim or zineb.

8. The method as claimed in claim 7, wherein the concentration of the hormone in the hormone sterilization solution is 0.0075-0.015 mg/mL, and the mass concentration of the sterilizing agent is 0.5-1%.

9. The method according to claim 7 or 8, wherein the time for root dipping treatment is 10 to 60 seconds.

10. The method of claim 6, wherein the robust growth characteristics include leaf size and stretch, bright green color, and being dry, straight, tough and tough.