Soil-borne disease detection method based on soil microbial community

Abstract

The invention discloses a soil-borne disease detection method based on a soil microbial community, which comprises the following steps: selecting an area for measuring the detection of the soil microbial community, determining the size of the selected area, delimiting the selected area, observing the plant growth state of the selected area and the actual condition of soil, and performing preliminary judgment; the method comprises the following steps: formulating a soil sampling mode, calibrating an initial position of soil sampling in a selected area, sequentially sampling the soil from the initial position, independently storing the sampled soil by adopting sample boxes, marking the sample boxes, and sequentially detecting and analyzing the types and the quantity of microorganisms in the soil of each sample box. According to the soil-borne disease detection method for the soil microflora, diversified soil sampling modes are adopted, comprehensive and comprehensive sampling of soil is achieved, multi-angle analysis and judgment are conducted through observation, microbiological analysis and experimental detection, and the soil-borne diseases of the soil microflora are accurately detected.

Description

Technical Field

[0001] The present invention relates to the technical field of soil-borne disease detection methods, and particularly to a soil-borne disease detection method based on soil microbial communities. Background Art

[0002] Soil refers to a layer of loose material on the earth's surface, composed of various granular minerals, organic matter, moisture, air, microorganisms, etc., and can grow plants. Soil is composed of minerals weathered from rocks, organic matter produced by the decomposition of plant and animal residues and microorganisms, soil organisms (solid phase substances), as well as moisture (liquid phase substances), air (gas phase substances), oxidized humus, etc.

[0003] Solid substances include soil minerals, organic matter, nutrients obtained after light-inhibiting and sterilizing microorganisms, etc. Liquid substances mainly refer to soil moisture, and the gas is the air existing in soil pores. These three types of substances in the soil constitute a contradictory unity. They are interconnected and restrict each other, providing essential living conditions for crops and being the material basis of soil fertility.

[0004] The substances in the soil can be generally summarized into three parts: the solid part, the liquid part, and the gas part (solid phase, liquid phase, and gas phase).

[0005] Soil minerals are mineral particles of different sizes (sand grains, soil grains, and colloidal particles) formed by the weathering of rocks. There are many types of soil minerals with complex chemical compositions. It directly affects the physical and chemical properties of the soil and is one of the important sources of crop nutrients.

[0006] The solid soil particles composed of minerals and humus are the main body of the soil, accounting for about 50% of the soil volume. The pores between the solid particles are occupied by gas and moisture.

[0007] Most of the gas in the soil is oxygen, nitrogen, etc. entering from the atmosphere, and a small part is carbon dioxide, water vapor, etc. produced by the life activities in the soil. The moisture in the soil mainly enters the soil from the surface, including many dissolved substances.

[0008] There are a large number of microorganisms in the soil. Diseases will also occur in the soil under the action of microorganisms. Some soil diseases gradually expand under the influence of external destructive factors. In order to understand the situation of soil-borne diseases in the soil microbial community, it is necessary to detect the soil. However, at present, the detection of soil diseases generally uses sampling observation or microbial analysis. However, such soil disease inspections are relatively one-sided and cannot comprehensively understand the disease situation of the soil. Therefore, an improved technology is urgently needed to solve this problem existing in the prior art. Summary of the Invention

[0009] The object of the present invention is to provide a soil-borne disease detection method based on soil microbial communities, which can achieve comprehensive and integrated sampling of soil, and conduct multi-angle analysis and judgment through observation, microbial analysis and experimental detection, so as to accurately detect soil-borne diseases of soil microbial communities, and solve the problems raised in the above-mentioned background technology.

[0010] To achieve the above object, the present invention provides the following technical solution: A soil-borne disease detection method based on soil microbial communities, the soil-borne disease detection method of the soil microbial communities includes:

[0011] S1. Select the area for measuring soil microbial community detection, determine the size of the selected area, and delimit the selected area;

[0012] S2. Observe the plant growth state and the actual situation of the soil in the delimited area, and make a preliminary judgment;

[0013] S3. Develop a soil sampling method, calibrate the initial position of soil sampling in the selected area, and sequentially conduct soil sampling starting from the initial position. The sampled soil is independently stored in a sample box, and the sample box is marked;

[0014] S4. Sequentially detect and analyze the types and quantities of microorganisms in the soil of each sample box, and isolate and identify the microorganisms and pathogens in the soil samples;

[0015] S5. Measure and analyze the small animals and microorganisms in the soil, and statistically analyze the measured data.

[0016] Preferably, when delimiting the area in S1, a circular range, a square range or a triangular range is used to enclose the area;

[0017] When sequentially selecting the area, the same enclosing method is used to enclose the land area.

[0018] Preferably, in S2, observe the plant growth state of the selected area, observe the number of plants growing inside the selected area, the types of plants and the plant growth state;

[0019] The plant growth state includes the height of the plant, the leaf state of the plant, and the pest state of the plant;

[0020] The actual situation of the soil includes the water content, minerals, organic matter and microorganisms in the soil, and the proportion of water content, minerals, organic matter and microorganisms in each soil sample is detected respectively.

[0021] Preferably, the soil sampling method is sequential sampling in a straight line. Starting from the calibrated location in the selected area as the initial sampling position, sequential sampling is carried out. The mass of each sampled soil is 120g - 130g, and each soil sample is sealed and stored in a transparent sealed box;

[0022] The external of the sample box is marked with the sampling time, the mass of the sampled sample, and the sampling location.

[0023] Preferably, in S4, after diluting the soil sample, it is spread on an agar medium, and the growing colonies are observed and counted;

[0024] After diluting the soil sample, a smear or suspension is prepared and observed using an optical microscope or an electron microscope to provide morphological information of the microorganisms.

[0025] Preferably, the data measured for each sample are statistically analyzed, and the various data in each sample are compared.

[0026] Preferably, in S3, a cylindrical sampling tube is used for soil sampling, and the digging depth of each soil sample is the same.

[0027] Preferably, in S4, the soil is placed in a separation sieve to break up the soil, and the substances in the broken-up soil are counted and classified.

[0028] Compared with the prior art, the beneficial effects of the present invention are:

[0029] (1) Each soil sample is sealed and stored in a transparent sealed box. The sampled soil is independently stored in a sample box, and the sample box is marked. The external of the sample box is marked with the sampling time, the mass of the sampled sample, and the sampling location. The transparent sample sealed box not only has good airtightness, but also can avoid the loss of soil moisture and the loss of microorganisms in the sample;

[0030] (2) By using equidistant continuous sampling, the specific situation of soil diseases and the spread range can be observed;

[0031] (3) After diluting the soil sample, a smear or suspension is prepared and observed using an optical microscope or an electron microscope to provide morphological information of the microorganisms. The number and types of microorganisms in the soil are detected through an electron microscope, so as to judge the general situation of the distribution of microorganisms in the soil, and thus indirectly judge the soil-borne disease situation of the soil microbial community;

[0032] (4) The method for detecting soil-borne diseases of the soil microbial community adopts a diversified soil sampling method to achieve comprehensive and comprehensive sampling of the soil, and conducts multi-angle analysis and judgment through observation, microbial analysis, and experimental detection to accurately detect the soil-borne diseases of the soil microbial community. Brief Description of the Drawings

[0033] Figure 1 It is a flow chart of the method for detecting soil-borne diseases based on the soil microbial community of the present invention. Detailed Description of the Embodiments

[0034] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0035] Please refer to Figure 1 , the present invention provides a technical solution: a method for detecting soil-borne diseases based on the soil microbial community, and the method for detecting soil-borne diseases of the soil microbial community includes:

[0036] S1. Select the area for measuring the soil microbial community detection, determine the size of the selected area, and demarcate the selected area.

[0037] S2. Observe the plant growth status and the actual situation of the soil in the demarcated area for a preliminary judgment.

[0038] S3. Develop a soil sampling method, mark the initial position of soil sampling in the selected area, and sequentially conduct soil sampling starting from the initial position. The sampled soil is independently stored in a sample box, and the sample box is marked.

[0039] S4. Sequentially detect and analyze the types and quantities of microorganisms in each sample box of soil, and isolate and identify the microorganisms and pathogens in the soil samples.

[0040] S5. Measure and analyze the small animals and microorganisms in the soil, and statistically analyze the measured data.

[0041] Select the area for measuring the soil microbial community detection, determine the size of the selected area, demarcate the selected area. When demarcating the area, use a circular range, a square range, or a triangular range to enclose the area. When sequentially selecting the area, use the same enclosing method to enclose the land area.

[0042] Example 1

[0043] Select the area for measuring soil microbial community detection, determine the size of the selected area, delimit the selected area. When delimiting the area, use a circle to enclose the range. Determine the starting point of sampling inside the enclosed range, and conduct circular sampling within the circular enclosed range, with equal-distance circular sampling outward from the circular position.

[0044] Example 2

[0045] Select the area for measuring soil microbial community detection, determine the size of the selected area, delimit the selected area. When delimiting the area, use a square to enclose it. Inside the enclosed range, use a diagonal straight-line sampling method for sampling, with a radius distance interval of 30 cm for each sampling loop.

[0046] Example 3

[0047] Select the area for measuring soil microbial community detection, determine the size of the selected area, delimit the selected area. When delimiting the area, use an equilateral triangle to enclose the range. Inside the enclosed range, use each corner as the starting point and sample along the sides of the triangle, with a sampling interval of 30 cm each time.

[0048] Example 4

[0049] Select the area for measuring soil microbial community detection, determine the size of the selected area, delimit the selected area. When delimiting the area, use a square to enclose the range. Conduct cross sampling inside the enclosed orientation, with a sampling interval of 30 cm each time.

[0050] Soil sampling is carried out using a cylindrical sampling tube, and the digging depth of each soil sample is the same. Place the soil in a sieve to break up the soil, and count and classify the substances in the broken-up soil.

[0051] By using equally spaced continuous sampling, the specific situation of soil diseases and the spread range can be observed.

[0052] Observe the plant growth status in the selected area, observe the number of plants growing inside the selected area, the types of plants, and the growth status of plants. The growth status of plants includes the height of plants, the leaf status of plants, and the pest status of plants. Statistically classify the plant species in the selected area, observe the growth status of the same type or the same species of plants, sort the counted plants according to the number of plants, and observe the growth status of each plant, analyzing the leaves, roots, and tendrils of plants. Different plants have different requirements for the soil growth environment. Judge the distribution of soil diseases and the distribution of soil diseases through the types of growing plants and the specific situation of plant growth.

[0053] The actual conditions of the soil include the water content, minerals, organic matter and microorganisms in the soil. The proportions of the water content, minerals, organic matter and microorganisms in each soil sample are detected respectively, and the plant growth status and the actual conditions of the soil in the delineated area are observed for a preliminary judgment.

[0054] Formulate the soil sampling method. The soil sampling method is sequential sampling in a straight line. Starting from the calibrated location in the selected area as the initial sampling position, sequential sampling is carried out. Calibrate the initial position of soil sampling in the selected area, and start sequential soil sampling from the initial position. The mass of each sample taken is 120g - 130g. Each soil sample is stored sealed in a transparent sealed box. The sampled soil is stored independently in a sample box, and the sample box is marked. The external of the sample box is marked with the sampling time, the mass of the sampled sample, and the sampling location. The transparent sample sealed box not only has good airtightness, but also can avoid the loss of soil water seal and the loss of microorganisms in the sample.

[0055] Sequentially detect and analyze the types and quantities of microorganisms in the soil of each sample box, isolate and identify the microorganisms and pathogens in the soil samples. After diluting the soil samples, spread them on the agar medium, observe and count the growing colonies. After diluting the soil samples, prepare smears or suspensions and observe them using an optical microscope or an electron microscope to provide morphological information of the microorganisms. Detect the quantity and types of microorganisms in the soil through an electron microscope, so as to judge a general situation of the distribution of microorganisms in the soil, and indirectly judge the soil-borne diseases of the soil microbial community.

[0056] Sequentially detect and analyze the types and quantities of microorganisms in the soil of each sample box, isolate and identify the microorganisms and pathogens in the soil samples, and accurately judge whether there are diseases in the soil. Common analysis methods include polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), etc.

[0057] Statistically analyze the data measured for each sample, and compare the various data in each sample. Measure and analyze the small animals and microorganisms in the soil, statistically analyze the measured data, and statistically analyze the data detected in the soil of each sample and make a table or pie chart to intuitively judge the soil-borne disease situation of the soil microbial community from the actual data.

[0058] The method for detecting soil-borne diseases of the soil microbial community adopts a diversified soil sampling method to achieve comprehensive and integrated sampling of the soil, and conducts multi-angle analysis and judgment through observation, microbial analysis and experimental detection to accurately detect the soil-borne diseases of the soil microbial community.

[0059] Although embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A soil-borne disease detection method based on soil microbial communities, characterized in that: The soil-borne disease detection method of the soil microbial community comprises: S1. Select an area for measuring soil microbial community detection, determine the size of the selected area, and demarcate the selected area; S2. Observe the plant growth status and soil conditions in the designated area and make a preliminary judgment; S3. Formulate a soil sampling method, mark the initial position of soil sampling in the selected area, and start sampling the soil from the initial position in sequence. The sampled soil is stored independently in a sample box, and the sample box is marked; S4. Detect and analyze the types and quantities of microorganisms in the soil of each sample box in turn, and separate and identify the microorganisms and pathogens in the soil samples; S5. Measure and analyze small animals and microorganisms in the soil, and statistically analyze the measured data.

2. The method for detecting soil-borne diseases based on soil microbial communities according to claim 1, characterized in that: When demarcating the area in S1, a circular area, a square area or a triangular area is used; When selecting areas one by one, the same method of enclosing the land is used to enclose the area.

3. The method for detecting soil-borne diseases based on soil microbial communities according to claim 1, characterized in that: The S2 observes the growth status of plants in the selected area, and observes the number of plants growing in the selected area, the types of plants, and the growth status of plants; The plant growth status includes plant height, plant leaf status, and plant pest status; The actual conditions of the soil include the water content, minerals, organic matter and microorganisms in the soil, and the proportions of water content, minerals, organic matter and microorganisms in each soil sample are detected respectively.

4. The method for detecting soil-borne diseases based on soil microbial communities according to claim 1, characterized in that: The soil sampling method is linear sampling, starting from the marked location of the selected area as the initial sampling location, and the mass of each sample is 120g-130g, and each soil sample is sealed and stored in a transparent sealed box; The outside of the sample box is marked with the sampling time, the sample quality, and the sampling location.

5. The method for detecting soil-borne diseases based on soil microbial communities according to claim 1, characterized in that: The S4 dilutes the soil sample, spreads it on an agar medium, and observes and counts the growing colonies; After diluting the soil sample, a smear or suspension is prepared and observed using an optical microscope or electron microscope to provide morphological information of the microorganisms.

6. The method for detecting soil-borne diseases based on soil microbial communities according to claim 1, characterized in that: The data measured for each sample are statistically analyzed, and the data in each sample are compared.

7. The method for detecting soil-borne diseases based on soil microbial communities according to claim 1, characterized in that: The soil sampling in S3 is performed using a cylindrical sampling tube, and the digging depth of each soil sample is the same.

8. The method for detecting soil-borne diseases based on soil microbial communities according to claim 1, characterized in that: The soil in S4 is placed in a separation sieve to break up the soil, and the substances in the broken up soil are counted and classified.

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