A culture device and experimental method for monitoring the interaction of soil animals and microorganisms with plant root systems
By designing a culture device and experimental method suitable for the interaction between soil animals and microorganisms and plant roots, and using rough-surfaced microporous hollow silica beads and culture bottles with specific structures, the problem of real-time observation and sampling in the study of the interaction between soil animals and microorganisms and plant roots was solved, and the real-time performance and repeatability of the experiment were improved.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- NANJING INST OF ENVIRONMENTAL SCI MINIST OF ECOLOGY & ENVIRONMENT OF THE PEOPLES REPUBLIC OF CHINA
- Filing Date
- 2026-03-20
- Publication Date
- 2026-05-29
AI Technical Summary
In existing research on the interaction between soil animals and microorganisms and plant roots, the interaction phenomena are not visible, the interaction period is uncontrollable, the culture conditions are complex, there are many interfering factors, real-time observation and sampling are not possible, and the experimental conditions have poor repeatability.
A culture device and experimental method for monitoring the interaction between soil animals and microorganisms and plant roots were developed. Rough-surfaced microporous hollow silica beads were used as the culture medium. The culture flasks with umbrella-shaped or inverted V-shaped structures were designed. Combined with artificial climate chambers and aseptic operation, real-time observation and sampling were achieved, sample contamination was prevented, and experimental repeatability was improved.
It achieves uniform distribution and visualization of plant roots, supports normal plant growth, facilitates real-time sampling and detection, improves the efficiency of real-time observation and sampling in experiments, reduces sample contamination, and improves the repeatability of experimental conditions.
Smart Images
Figure CN122095989A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the technical field of plant growth and cultivation equipment, specifically relating to a cultivation device and experimental method for monitoring the interaction between soil animals and microorganisms and plant roots. Background Technology
[0002] Soil animals and soil microorganisms are living organisms that inhabit the soil. The former mainly includes invertebrates, including annelids, arthropods, mollusks, nematodes, and protozoa. The latter includes groups such as bacteria, actinomycetes, fungi, and algae.
[0003] Soil animals and soil microorganisms play a crucial role not only in rock weathering and the formation of primary soils, but also in soil growth and development, the formation and evolution of soil fertility, and the nutrient supply to higher plants. Furthermore, they interact with plant roots during plant growth, such as through herbivorous nematodes, rhizobia, and pathogens.
[0004] Currently, there are many difficulties in the study of the interaction between soil animals, soil microorganisms and plant roots, especially the fact that the interaction phenomenon is not visible, the interaction period is uncontrollable, the culture conditions are complex, there are many interfering factors, and real-time sampling is not possible.
[0005] To address the problems of existing research on the interaction between plant roots and soil animals and microorganisms, such as the inability to observe and sample in real time and poor repeatability of experimental conditions, it is an urgent problem for those skilled in the art to solve how to provide a culture device and experimental method for monitoring the interaction between soil animals and microorganisms and plant roots. Summary of the Invention
[0006] The main objective of this invention is to provide a cultivation device and experimental method for monitoring the interaction between soil animals and microorganisms and plant roots, thereby solving the aforementioned technical problems. This device and method allow for real-time observation and sampling of the monitored plants, minimizing sample contamination, and also addressing issues such as poor repeatability of experimental conditions. It facilitates sampling and testing by operators during experiments, making it more convenient to use.
[0007] To achieve the above objectives, the present invention adopts the following technical solution: A culture device for monitoring the interaction between soil animals and microorganisms and plant roots includes a culture bottle body, each culture bottle body containing a culture medium, and each culture bottle body having multiple exchange ports at its bottom end, each exchange port being covered with a cap.
[0008] Furthermore, the culture medium uses rough-surfaced microporous hollow silicon beads with a diameter of 0.5cm-1cm, and the material is silicon dioxide.
[0009] Furthermore, when the plant is a taproot plant, the main body of the culture bottle has an umbrella-shaped structure.
[0010] Furthermore, when the plant is a fibrous root system, the main body of the culture bottle has an inverted V-shaped structure.
[0011] An experimental method for monitoring the interaction between soil animals and microorganisms and plant roots includes the following steps: S1, Plant preparation: Select the appropriate plant varieties, disinfect the seed surface and germinate in a sterile environment, and transfer the plants to the corresponding culture bottle body according to their different root systems for cultivation. S2, Soil biological inoculation, involves injecting soil animals or microorganisms into the rhizosphere; S3, Cultivation and Management: Place the plants and the main body of the culture bottle in an artificial climate chamber. Adjust the light duration, day and night temperature, and relative humidity according to the experimental requirements. In addition, spray and irrigate with nutrient solution regularly according to the different needs of different plants. S4, growth monitoring, after inoculation, monitors the interaction between soil organisms and plant roots from multiple perspectives at various stages of plant growth. S5, real-time sampling, sampling the sample at different times of interaction between the two according to experimental needs; S6. After sampling and processing, the plants are put back into the culture bottle, and culture medium and culture solution are added to continue the culture, which facilitates continuous research.
[0012] Furthermore, in step S1, the mouth of the culture bottle body is sealed with double-layered sterile black gauze to maintain a sterile environment inside the culture bottle body, and the entire culture bottle body is covered with a black bag or placed in a dark box to prevent the root system from being exposed to light.
[0013] Furthermore, in step S5, the cover at the exchange port is opened during sampling to discharge the culture medium and culture solution. Substances in the culture medium or culture solution are extracted as needed. Then, the plant is removed, and samples are taken from the roots. After sampling, the plant is put back into the culture bottle body, and culture solution and culture medium are added again for continued cultivation. Subsequently, multiple sampling studies are conducted at different growth stages according to experimental requirements.
[0014] Compared with the prior art, the present invention has the following beneficial effects: In this invention, the plant root system is fully and evenly distributed and visualized through a cultivation device. In addition, the use of a culture medium to assist plant growth not only supports normal plant growth but also allows for convenient real-time sampling and testing of plant roots, resulting in higher efficiency. Furthermore, the experimental method in this invention solves the problems of existing studies on the interaction between plant roots and soil animals and microorganisms, such as the inability to observe and sample in real time, easy sample contamination, and poor repeatability of experimental conditions. This makes it easier for operators to perform sampling and testing during experiments, making it more convenient to use. Attached Figure Description
[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.
[0016] Figure 1 This is a schematic diagram of the experimental procedure.
[0017] Figure 2 This is a schematic diagram of the culture bottle structure for taproot plants.
[0018] Figure 3 This is a schematic diagram of the structure of a culture bottle for plants with fibrous roots.
[0019] Among them, 1-culture flask body, 2-culture medium, 3-exchange port. Detailed Implementation
[0020] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0021] Example 1, such as Figure 1-3 As shown, the present invention provides a culture device for monitoring the interaction between soil animals and microorganisms and plant roots, including a culture bottle body 1, a culture medium 2 is provided inside the culture bottle body 1, a plurality of exchange ports 3 are provided at the bottom of the culture bottle body 1, and a cover is provided at each of the exchange ports 3. The material of the culture bottle body 1 is glass or polycarbonate.
[0022] In this embodiment, the culture medium 2 uses rough-surfaced, microporous hollow silica beads with a diameter of 0.5cm-1cm. The material is silica, and the beads can be sterilized by high-temperature and high-pressure sterilization or by soaking in a 10% sodium hypochlorite solution before use. The hollow and microporous structure of the silica beads can retain air or moisture, providing a better growth environment for soil animals and microorganisms. The rough, uneven surface can increase the contact area between the individual silica beads and between the silica beads and the plant roots, and reduce damage to the plant roots during sampling.
[0023] In this embodiment, when the plant is a taproot plant, the main body 1 of the culture bottle has an umbrella-shaped structure.
[0024] In this embodiment, when the plant is a fibrous root plant, the main body 1 of the culture bottle has an inverted V-shaped structure.
[0025] An experimental method for monitoring the interaction between soil animals and microorganisms and plant roots includes the following steps: S1, Plant preparation: Select the appropriate plant varieties, disinfect the seed surface and germinate in a sterile environment. After about a week, transfer the plants to the corresponding culture bottle body 1 according to their different root systems for cultivation. Culture medium 2 and culture solution are prepared in advance in culture bottle body 1. Culture medium 2 is silica beads and culture solution is suitable for soil animals or soil microorganisms, such as Hoagland nutrient solution. S2, soil biological inoculation, involves injecting soil animals or microorganisms into the rhizosphere about one week after the main body of the plant transfer culture bottle is placed. In addition, when inoculation and transfer of soil animals, plants and microorganisms require aseptic conditions, they should be carried out on a sterile operating table to prevent contamination by other microorganisms. S3, Cultivation and Management: Place the plant and the main body of the culture bottle 1 in an artificial climate chamber or greenhouse. Adjust the light duration, day and night temperature, and relative humidity according to the experimental requirements. Spray and irrigate with nutrient solution at regular intervals according to the different needs of the plants. The artificial climate chamber or greenhouse can control the temperature, humidity, and light conditions. It is equipped with a temperature control system and a humidity controller, which can be adjusted according to different plants. Light can be adjusted by installing LED plant growth lights and shade nets. S4, growth monitoring, after inoculation, monitors the interaction between soil organisms and plant roots from multiple perspectives at various stages of plant growth; it monitors the culture environment by installing thermometers, hygrometers, pH meters, etc., and can also use cameras for auxiliary detection and recording.
[0026] S5 allows for real-time sampling, taking samples at different stages of interaction between the two components as needed for the experiment. It also allows for observation of soil animals and microbial infection status, counting of cysts, and observation of the interaction between microorganisms and roots using handheld and desktop microscopes. S6. After sampling and processing, the plant is put back into the culture bottle, and culture medium 2 is added to continue the culture, which facilitates continuous research.
[0027] In this embodiment, in step S1, the mouth of the culture bottle body 1 is sealed with double-layered sterile black gauze to maintain a sterile environment inside the culture bottle body 1. The entire culture bottle body 1 is covered with a black bag or placed in a dark box to prevent the root system from being exposed to light.
[0028] In this embodiment, during step S5, the cover at the exchange port 3 can be opened to discharge the culture medium 2 and culture solution. The substances in the culture medium 2 or culture solution can be extracted as needed. Then, the plant is taken out, and the sample on the root system is taken. After sampling, the plant is put back into the culture bottle body 1, and the culture solution and culture medium 2 are added again for continued cultivation. Afterwards, multiple sampling studies are conducted at different growth stages according to experimental requirements.
[0029] Example 2: Soybean cyst nematode (Heterodera glycines) is one of the most important pathogenic nematodes that harm soybean production worldwide. There are many difficulties in studying its biological characteristics, pathogenic mechanism and control technology. In particular, the infection process is invisible, the infection period is uncontrollable, it is difficult to detect and sample during the infection period, especially in the early stage of infection, the culture conditions are complicated, there are many interfering factors, real-time sampling is not possible, and standardization is difficult.
[0030] System preparation and sterilization: The cleaned culture bottle body 1 and culture medium 2 are sterilized by high-pressure steam. In a sterile operating table, the sterile germinated susceptible soybean plants are transferred into the umbrella-shaped culture bottle body 1, which is specially designed for taproot plants. The sterilized culture medium 2, i.e., silica beads, is filled in, and the culture solution, i.e., Hoagland nutrient solution, is added.
[0031] Host plant cultivation: Placed in an artificial climate chamber, the cultivation conditions are 14h / 10h photoperiod (day / night), day temperature 25±1℃, night temperature 20±1℃, and relative humidity 60-70%.
[0032] Nematode inoculation and culture: Collect purified second-instar larvae of soybean cyst nematodes and prepare a suspension at a certain concentration (e.g., 500-1000 larvae / mL). When the host plant seedlings are about 15 days old, inject the nematode suspension near the roots using a microsyringe. Place the main body of the culture bottle 1 back into the artificial climate chamber for continued culture.
[0033] Growth monitoring: After inoculation, the interaction between soybean cyst nematodes and soybean roots can be monitored from multiple angles at various stages of soybean growth, and various devices can be used to take pictures and record data.
[0034] Real-time sampling: As needed for the experiment, samples can be taken from the infected parts at different stages of the interaction between soybean cyst nematodes and soybean roots. During sampling, first open exchange port 3 to drain the silica beads and Hoagland nutrient solution, then remove the soybean and sample the infected parts of the roots. After sampling, the soybean can be placed back into the main body 1 of the culture bottle, and Hoagland nutrient solution and silica beads can be added again for continued cultivation. Multiple sampling studies can then be conducted at various growth stages as needed for the experiment.
[0035] Nematode collection: 35-45 days after inoculation, mature cysts begin to form on soybean roots. Silica beads and Hoagland nutrient solution can be drained, and the cysts on the roots and in the substrate can be collected. Larvae of all instars can also be collected from the silica beads and Hoagland nutrient solution.
[0036] Example 3: Wheat root rot is a global disease and one of the most important wheat diseases in China, occurring in all wheat-growing areas. It is widely distributed, especially in rainy years and humid regions. Infection often causes premature leaf withering, affecting grain filling and reducing thousand-grain weight. Infection of the ear can cause withered white ears, further impacting yield and quality. High seed-carrying rates reduce germination rates and cause root rot, severely affecting wheat emergence and seedling growth.
[0037] There are many challenges in studying the biological characteristics, pathogenic mechanisms, and control techniques of wheat root rot, including the fact that the infection process is invisible, the infection period is uncontrollable, it is difficult to detect and sample during the infection period, especially in the early stage, the culture conditions are complex, there are many interfering factors, real-time sampling is not possible, and standardization is difficult.
[0038] Activation and sporulation: Inoculate Bipolaris sorokiniana onto PDA plates and incubate at 28-30℃ for 5-7 days until the colonies mature and produce a large number of spores.
[0039] Preparation of spore suspension: Wash spores with sterile water or 0.05% Tween solution, filter through multiple layers of gauze or filter paper to remove hyphae, and adjust the concentration to 10–10 spores / mL using a hemocytometer (10 spores / mL is commonly used). Prepare and use immediately.
[0040] Host plant cultivation: Select susceptible or testable wheat varieties, and after one week of germination, transfer them to the main body 1 of a culture bottle with an inverted V-shaped structure specifically for fibrous root plants. Fill the bottle with sterilized culture medium 2, which is silica beads, and add Hoagland nutrient solution. Cultivate the seedlings to the required seedling age for inoculation (such as the one-leaf-one-heart stage or the three-leaf stage) to ensure that the wheat seedlings are healthy and grow uniformly.
[0041] Inoculation and Cultivation: Inject the prepared spore suspension into the vicinity of the roots using a syringe. Immediately after inoculation, cover the plant with a transparent plastic bag or cling film for 24-72 hours, maintaining a relative humidity of over 90%, which is crucial for successful infection. Place the plant in an environment of 25-28℃; darkness or low light conditions are more conducive to invasion. After approximately 24 hours, transition to a normal photocycle (e.g., 12 hours light / 12 hours darkness).
[0042] Disease observation and recording: Typical symptoms (browning of roots and stem base, spindle-shaped lesions on leaves, etc.) usually begin to appear 7-14 days after inoculation. Root samples can be taken for research at any time after inoculation, and subsequent cultivation and observation can continue as needed.
[0043] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on its differences from other embodiments. Similar or identical parts between embodiments can be referred to interchangeably. For the apparatus disclosed in the embodiments, since they correspond to the methods disclosed in the embodiments, the description is relatively simple; relevant parts can be referred to the method section.
[0044] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. A culture device for monitoring the interaction between soil animals and microorganisms and plant roots, characterized in that, The culture bottle includes a culture bottle body (1), and a culture medium (2) is provided inside the culture bottle body (1). Multiple exchange ports (3) are provided at the bottom of the culture bottle body (1), and a cover is provided at each of the exchange ports (3).
2. The culture device for monitoring the interaction between soil animals and microorganisms and plant roots according to claim 1, characterized in that, The culture medium (2) uses rough-surfaced microporous hollow silicon beads with a diameter of 0.5cm-1cm and is made of silicon dioxide.
3. The culture device for monitoring the interaction between soil animals and microorganisms and plant roots according to claim 2, characterized in that, When the plant is a taproot plant, the main body (1) of the culture bottle is an umbrella-shaped structure.
4. The culture device for monitoring the interaction between soil animals and microorganisms and plant roots according to claim 2, characterized in that, When the plant is a fibrous root system, the main body (1) of the culture bottle is an inverted V-shaped structure.
5. An experimental method for monitoring the interaction between soil animals and microorganisms and plant roots, implemented using the culture apparatus described in any one of claims 1-4, characterized in that, Includes the following steps: S1, Plant preparation: Select the appropriate plant varieties, disinfect the seed surface and germinate in a sterile environment, and transfer the plants to the corresponding culture bottle body (1) according to their different root systems for cultivation. S2, Soil biological inoculation, involves injecting soil animals or microorganisms into the rhizosphere; S3, cultivation and management: place the plant and the main body of the culture bottle (1) in an artificial climate chamber, adjust the light time, day and night temperature and relative humidity according to the experimental needs, and spray and irrigate nutrient solution regularly according to the needs of different plants. S4, growth monitoring, after inoculation, monitors the interaction between soil organisms and plant roots from multiple perspectives at various stages of plant growth. S5, real-time sampling, sampling the sample at different times of interaction between the two according to experimental needs; S6. After sampling and processing, the plant is put back into the culture bottle, and culture medium and culture medium (2) are added to continue the culture, so as to facilitate continuous research.
6. The experimental method for monitoring the interaction between soil animals and microorganisms and plant roots according to claim 5, characterized in that, In step S1, the mouth of the culture bottle body (1) is sealed with double-layer sterile black gauze to maintain a sterile environment inside the culture bottle body (1). The entire culture bottle body (1) is covered with a black bag or placed in a dark box to prevent the root system from being exposed to light.
7. The experimental method for monitoring the interaction between soil animals and microorganisms and plant roots according to claim 5, characterized in that, In step S5, when sampling, open the cover at the exchange port (3) to discharge the culture medium (2) and culture solution. Extract the substances in the culture medium (2) or culture solution as needed, then take out the plant and take samples from the root system. After sampling, put the plant back into the culture bottle body (1) and add culture solution and culture medium (2) again to continue the culture. Then, according to the experimental requirements, conduct multiple sampling studies at different growth stages.