Device and method for researching interaction of carbon nutrition and mineral nutrient elements of crops

Abstract

The invention discloses a device and a method for researching interaction of carbon nutrition and mineral nutrient elements of crops, the device comprises a culture dish and a middle cover plate, the culture dish comprises a dish body and a dish cover, the middle cover plate is provided with an upward support and a downward foot stool, the foot stool of the middle cover plate is supported in the dish body of the culture dish, and the dish cover is provided with a water inlet and a water outlet. And the bracket of the middle cover plate upwards supports the dish cover of the culture dish. According to the device and the method, photosynthesis can be well inhibited on the basis of not influencing plant photomorphological establishment, endogenous carbon nutrient substances of crops can be consumed in a short time, and the plants can quickly respond to exogenous carbon and nitrogen supply. Meanwhile, the device and the method can be expanded and applied to research on interaction of carbon nutrition and other mineral nutrient elements of other crops such as wheat, soybean and corn.

Description

Technical Field

[0001] This invention relates to the field of biology, and more specifically, to an apparatus and method for studying the interaction between crop carbon nutrition and other mineral nutrients. Background Technology

[0002] As autotrophic organisms, plants convert carbon dioxide and water into organic carbohydrates necessary for their growth through photosynthesis in their leaves, while simultaneously absorbing and utilizing other mineral nutrients from the external environment and soil through their roots. Extensive research in recent decades has revealed a complex interplay of regulatory and balancing mechanisms between photosynthetically fixed carbon nutrients and their products (such as sucrose, glucose, and trehalose-6-phosphate) and mineral nutrients absorbed and utilized by roots (such as nitrogen, sulfur, phosphorus, potassium, calcium, and magnesium), thus coordinating plant growth and physiological functions. Therefore, systematically elucidating the molecular mechanisms of the interaction between carbon nutrients and other mineral nutrients is crucial for improving crop photosynthetic efficiency and the efficiency of mineral nutrient absorption and utilization, thereby guiding the cultivation of high-yield, low-consumption, and environmentally friendly new crop varieties.

[0003] Because plants can synthesize carbon nutrients autonomously, precise control of carbon nutrition is crucial for studying its interactions with other mineral nutrients. However, there are currently no well-established and systematic research devices and methods for studying the interactions of carbon nutrition with other mineral nutrients, especially for crops. One challenge is that because plants can synthesize carbohydrates through photosynthesis, it is impossible to precisely control the concentration of carbon (i.e., sugar concentration) in plants. Previous solutions involved using shading to reduce photosynthesis and inhibit sugar synthesis, but this method also inhibits photomorphogenesis, thus having certain drawbacks. Another challenge is that the cotyledons or endosperm of crops store large amounts of carbohydrates such as starch, and their internal nutrient supply can sustain their growth for a long time. Therefore, they are not sensitive to changes in external nutrient supply, making it difficult to disrupt their internal carbon-nitrogen balance in a short period of time. Summary of the Invention

[0004] To address the two challenges mentioned above, this invention presents a novel research device and method adapted for studying carbon-nitrogen interactions, using rice as the research subject.

[0005] This invention provides an apparatus for precisely controlling the carbon nutrient supply to crops and studying the interaction between carbon and mineral nutrients. The apparatus includes a petri dish and an intermediate cover plate. The petri dish includes a dish body and a lid. The intermediate cover plate has an upward-facing support and a downward-facing leg. The leg of the intermediate cover plate supports the petri dish body, and the support of the intermediate cover plate supports the petri dish lid. In use, seeds are placed under the intermediate cover plate, and the culture solution is used to completely submerge the seeds or plants, ensuring that the crop seedlings are completely immersed in the liquid culture medium. This achieves the purpose of inhibiting plant photosynthesis while ensuring normal photomorphogenesis without causing stress such as waterlogging or hypoxia.

[0006] Furthermore, the intermediate cover plate is provided with multiple holes, which are evenly or unevenly distributed on the intermediate cover plate.

[0007] Furthermore, the cross-sectional shape of the culture dish is circular, square, rectangular, or irregular; the height of the dish body is 5-100 mm, the height of the dish lid is 5-100 mm, and the area of ​​the dish body and the dish lid is 2826-160000 mm². 2 .

[0008] Furthermore, the shape of the intermediate cover plate is adapted to the shape of the petri dish. The intermediate cover plate is placed inside the dish and the dish lid is closed. The intermediate cover plate divides the internal space of the petri dish into an upper and lower part, with the height of the lower part being the length of the legs. The thickness of the intermediate cover plate is 0.3-3 mm. The legs are vertically bent downwards along the edge of the intermediate cover plate, and there are 4-8 legs. The supports are vertically bent upwards along the edge of the intermediate cover plate, and there are 2-8 supports. The length of the legs is 1-100 mm and the width is 3-20 mm. The length of the supports is 5-100 mm and the width is 3-20 mm. The pore diameter is 1-10 mm, and the pore density is 0.1-5 pores / cm³. 2 Different shapes, areas, and heights of intermediate cover plates are provided according to the shape, area, and height of the petri dishes used. The size is selected according to the different petri dish sizes and the size of the crop seedlings. The perforations can reduce air bubbles caused by surface tension of the nutrient solution to a certain extent. The area of ​​the intermediate cover plate is determined according to the petri dish body, so that the cover plate can be inserted into the petri dish body and is 1-5 mm away from the dish wall, so that it can be fixed in the petri dish. The legs can leave space between the cover plate and the bottom to prevent the seeds from being crushed and damaged. The support can hold the lid against the dish to prevent it from being pushed up during seedling growth.

[0009] Furthermore, the culture dish is made of a highly transparent polymer material or transparent glass material, wherein the highly transparent polymer material is polycarbonate or polystyrene. The middle cover plate is made of a highly transparent polymer material such as polycarbonate or polystyrene, or transparent glass material.

[0010] This invention provides a method for studying the interaction between crop carbon nutrition and mineral nutrients, including the step of immersing seeds with part of the endosperm removed in a culture solution below an intermediate cover plate using the aforementioned apparatus, wherein the culture solution is a nutrient solution containing different proportions of carbon and other mineral nutrients.

[0011] Furthermore, the amount of culture medium is selected based on the area of ​​the petri dish, the height of the middle cover stand, and the crop seedlings, so that the height of the culture medium in the petri dish just covers the entire seedling; the culture medium is changed every 2-4 days.

[0012] Furthermore, the other nutrients are nitrogen or mineral elements; the culture is carried out under sterile conditions.

[0013] Furthermore, the crop is rice, wheat, corn, or soybean.

[0014] Furthermore, the crop is rice, and the endosperm is removed in a ratio of 2 / 3.

[0015] The apparatus and method of this invention can effectively suppress photosynthesis without affecting plant photomorphogenesis, while simultaneously consuming endogenous carbon nutrients in crops in a short period, enabling plants to rapidly respond to exogenous carbon and nitrogen supplies. Furthermore, this apparatus and method can be extended to studies of the interaction between carbon nutrition and other mineral nutrients in other crops such as wheat, soybeans, and corn.

[0016] In summary, this invention establishes an apparatus and method for studying the interaction between crop carbon nutrition and other mineral nutrients, laying the foundation for the analysis of molecular mechanisms and molecular breeding in future scientific research. Attached Figure Description

[0017] Figure 1 This is a structural example diagram of a device for precisely controlling crop carbon nutrient supply and studying the interaction between carbon and mineral nutrients, where A is a three-dimensional view, B is a longitudinal sectional view, and C is a transverse sectional view.

[0018] Figure 2 This is a structural example diagram of another device used for precisely controlling crop carbon nutrient supply and studying the interaction between carbon and mineral nutrients, where A is a three-dimensional view, B is a longitudinal sectional view, and C is a transverse sectional view.

[0019] Figure 3 This is a schematic diagram of the use of the above-mentioned device to study the carbon and nitrogen balance of rice, where A is a schematic diagram and B is a photograph of the cultivation state.

[0020] Figure 4 To observe the phenotypic characteristics of rice seedlings with different proportions of endosperm removed, the scale bar was 2 cm.

[0021] Figure 5 To set up a glucose concentration gradient and determine the sugar supply concentration, the scale bar is 2 cm.

[0022] Figure 6 Phenotypic observation and physiological measurement of rice seedlings under different carbon and nitrogen concentrations. Scale bar: 2 cm.

[0023] Figure 7 For phenotypic statistics of transitioning from carbon-nitrogen imbalance to carbon-nitrogen balance conditions, the scale bar is 2 cm.

[0024] Explanation of reference numerals in the attached diagram: 1. Lid of the dish, 2. Middle cover plate, 3. Dish body, 21. Support, 22. Leg, 23. Hole. Detailed Implementation

[0025] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0026] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0027] Unless otherwise specified, the quantitative experiments in the following examples are all repeated three times, and the results are averaged. Invention Overview This invention provides an apparatus for precisely controlling the carbon nutrient supply to crops and studying the interaction between carbon and mineral nutrients. The apparatus includes a petri dish and an intermediate cover plate. The petri dish includes a dish body and a lid. The intermediate cover plate has an upward-facing support and a downward-facing leg. The leg of the intermediate cover plate supports the petri dish body, and the support of the intermediate cover plate supports the petri dish lid. In use, seeds are placed under the intermediate cover plate, and the culture solution is used to completely submerge the seeds or plants, ensuring that the crop seedlings are completely immersed in the liquid culture medium. This achieves the purpose of inhibiting plant photosynthesis while ensuring normal photomorphogenesis without causing stress such as waterlogging or hypoxia.

[0029] Furthermore, the intermediate cover plate is provided with multiple holes, which are evenly or unevenly distributed on the intermediate cover plate.

[0030] Furthermore, the cross-sectional shape of the culture dish is circular, square, rectangular, or irregular; the height of the dish body is 5-100 mm, the height of the dish lid is 5-100 mm, and the area of ​​the dish body and the dish lid is 2826-160000 mm². 2 .

[0031] Furthermore, the shape of the intermediate cover plate is adapted to the shape of the petri dish. The intermediate cover plate is placed inside the dish and the dish lid is closed. The intermediate cover plate divides the internal space of the petri dish into an upper and lower part, with the height of the lower part being the length of the legs. The thickness of the intermediate cover plate is 0.3-3 mm. The legs are vertically bent downwards along the edge of the intermediate cover plate, and there are 4-8 legs. The supports are vertically bent upwards along the edge of the intermediate cover plate, and there are 2-8 supports. The length of the legs is 1-100 mm and the width is 3-20 mm. The length of the supports is 5-100 mm and the width is 3-20 mm. The pore diameter is 1-10 mm, and the pore density is 0.1-5 pores / cm³. 2 Different shapes, areas, and heights of intermediate cover plates are provided according to the shape, area, and height of the petri dishes used. The size is selected according to the different petri dish sizes and the size of the crop seedlings. The perforations can reduce air bubbles caused by surface tension of the nutrient solution to a certain extent. The area of ​​the intermediate cover plate is determined according to the petri dish body, so that the cover plate can be inserted into the petri dish body and is 1-5 mm away from the dish wall, so that it can be fixed in the petri dish. The legs can leave space between the cover plate and the bottom to prevent the seeds from being crushed and damaged. The support can hold the lid against the dish to prevent it from being pushed up during seedling growth.

[0032] Furthermore, the culture dish is made of a highly transparent polymer material or transparent glass material, wherein the highly transparent polymer material is polycarbonate or polystyrene. The middle cover plate is made of a highly transparent polymer material such as polycarbonate or polystyrene, or transparent glass material.

[0033] This invention provides a method for studying the interaction between crop carbon nutrition and mineral nutrients, including the step of immersing seeds with part of the endosperm removed in a culture solution below an intermediate cover plate using the aforementioned apparatus, wherein the culture solution is a nutrient solution containing different proportions of carbon and other mineral nutrients.

[0034] Furthermore, the amount of culture medium is selected based on the area of ​​the petri dish, the height of the middle cover stand, and the crop seedlings, so that the height of the culture medium in the petri dish just covers the entire seedling; the culture medium is changed every 2-4 days.

[0035] Furthermore, the other nutrients are nitrogen or mineral elements; the culture is carried out under sterile conditions.

[0036] Furthermore, the crop is rice, wheat, corn, or soybean.

[0037] Furthermore, the crop is rice, and the endosperm is removed in a ratio of 2 / 3.

[0038] Example 1 This invention uses an intermediate cover plate to immerse the plant in the nutrient solution, reducing carbon dioxide absorption and thus inhibiting photosynthesis, thereby minimizing the impact of endogenous sugar synthesis in the plant. In this invention... Figure 1 The structure of a square petri dish and its matching center cover is described. Figure 2 The text lists round petri dishes and their suitable middle covers. The appropriate petri dish and cover should be selected based on the size and growth stage of the crop seeds and seedlings. This includes the cover's shape, area, and thickness; the length, width, number, and position of the legs and supports; and the location, size, and density of the perforations. The edge of the cover should be approximately 1-3 mm from the inner diameter of the petri dish to ensure it fits snugly inside.

[0039] This invention, taking the study of carbon and nitrogen interactions in rice as an example, designs a petri dish and its compatible intermediate cover structure. The invention uses a circular petri dish as the culture container; the dish body is 20 mm high, 134 mm in inner diameter, and has an area of ​​approximately 14100 mm². 2 The petri dish lid 1 is 7 mm high and 143 mm in diameter. A central cover plate 2 is designed and manufactured according to the size of the petri dish; it is circular in shape, 128 mm in diameter, and 1 mm thick. The holes 23 are 4 mm in diameter with a spacing of 19 mm. The cover plate has four legs 22 and two supports 21, each 11 mm long and 10 mm wide. The central cover plate is securely fixed in the petri dish, leaving space between the cover and the bottom to prevent damage to the seeds. The supports hold the petri dish lid in place to prevent the seedlings from being pushed up during growth. The cover plate is made of polycarbonate (PC), with a light transmittance of approximately 88%. It can be autoclaved (121°C, 20 minutes) to ensure sterility during cultivation. PC plastic is relatively soft; the legs and supports are cut to size according to the crop seeds and the height of the petri dish before use. Its usage is as follows... Figure 3 As shown.

[0040] This invention provides a method for cultivating rice seedlings using the aforementioned carbon-nitrogen concentration ratio, comprising the following steps: 1) Remove the shells from the Nipponbare seeds, disinfect them by washing with sodium hypochlorite with an effective chlorine content of 2.5% for 40-50 minutes, place them in a 9cm round petri dish, add 6 mL of sterile water, soak them at 28℃ for 24 hours, and then germinate them at 37℃ for 24-48 hours. 2) Select rice seeds with uniform germination, remove 2 / 3 of the endosperm (approximately at the lower edge of the embryo), place them in a new 9 cm culture dish, add 8 mL of nutrient solution, and culture them under the conditions of 25-28℃ and light:dark = 12 h: 12 h. 3) After 3 days of cultivation, select seedlings with uniform growth of plumule and radicle, transfer them to 15 cm round dishes, add 40 mL of nutrient solution, cover with a perforated round cover plate, and continue cultivation. 4) On the seventh day of cultivation, the seedlings can be removed and their phenotypes observed.

[0041] Example 2 1. Material preparation 1) Plant material: Nipponbare rice seeds.

[0042] 2) Instruments and equipment: clean bench, forceps, scalpel, light incubator, sealing film, 9 cm culture dish (BIOFIL) for surface tension reduction, 15 cm culture dish (BIOFIL), 1 L filtration flask (BIOFIL), 1 L beaker, pipette, transparent perforated plate.

[0043] 3) Experimental reagents: sodium hypochlorite solution (available chlorine content 2-2.5%), sterile water, Kimura B nitrogen-free culture medium stock solution (Beijing Cooler Master Technology Co., Ltd.).

[0044] 2. Preparation of liquid culture medium 1) Preparation of nitrogen-free 1 / 2 Kimura B liquid medium: Add 500 μL of 1000× nitrogen-free macro- and micro-element mixture, 500 μL of 1000× iron salt, 500 μL of 1000× calcium salt, and 625 μL of 1M K2SO4 to a 1 L beaker, and bring the volume to 1 L. The pH is 5.7. Prepare nitrogen-free 1 / 2 Kimura B liquid medium and filter and sterilize it using a 1 L vacuum filtration flask.

[0045] 2) Preparation of nitrogen-added 1 / 2 Kimura B liquid medium: Add 500 μL of 1000× nitrogen-free macro- and micro-element mixture, 500 μL of 1000× iron salt, 500 μL of 1000× calcium salt, 1.25 mL of 1 M KNO3, and 1.25 mL of 0.5 M (NH4)2SO4 to a 1 L beaker, and adjust the volume to 1 L. Set the pH to 5.7 to prepare nitrogen-added 1 / 2 Kimura B liquid medium (total nitrogen concentration 2.5 mM). Filter and sterilize using a 1 L vacuum filtration flask.

[0046] 3) Preparation of culture medium: Using 1 / 2 nitrogen-added Kimura B liquid medium as a low-carbon, high-nitrogen culture medium (total carbon concentration 0 mM, total nitrogen concentration 2.5 mM), different amounts of glucose were added to obtain high-carbon, high-nitrogen culture media with different C:N ratios. For example, 2.7 g of glucose was added to 1 L of 1 / 2 nitrogen-added Kimura B liquid medium and mixed thoroughly to obtain a high-carbon, high-nitrogen culture medium (at this point, the total carbon concentration in the culture medium was 15 mM, and the total nitrogen concentration was 2.5 mM).

[0047] Using nitrogen-free 1 / 2 Kimura B liquid medium as a low-carbon, low-nitrogen culture medium (total carbon concentration 0 mM, total nitrogen concentration 0 mM), different amounts of glucose were added to obtain high-carbon, low-nitrogen culture media with different C:N ratios. For example, adding 2.7 g of glucose to 1 L of nitrogen-free 1 / 2 Kimura B liquid medium and mixing thoroughly yielded a high-carbon, low-nitrogen culture medium (at this point, the total carbon concentration in the culture medium was 15 mM, and the total nitrogen concentration was 0 mM).

[0048] 3. Seed washing 1) Remove the husks from the dried rice seeds, place them in a centrifuge tube, add 20-30 mL of 75% ethanol, and wash them on a shaker for 2 minutes; 2) Discard 75% ethanol, add sodium hypochlorite solution (available chlorine is 2-2.5%), and wash on a shaker for 40-50 min; 3) Discard the sodium hypochlorite solution in the clean bench and rinse with sterile water 4-5 times; 4) Place about 20 rice seeds into a sterile 9 cm culture dish with de-surface tension. 5) Add 6 mL of sterile water to each petri dish and seal with sealing film; 6) Place in an incubator at 28℃ overnight, then place in an incubator at 37℃ the next day to germinate; 7) After 24-48 hours of germination, both the plumule and radicle will emerge.

[0049] 4. Endosperm removal Crop seeds contain a large amount of nutrients, requiring the removal of their endosperm (or cotyledons) to deplete the crop's endogenous nutrients as much as possible. This allows for precise control of the carbon-nitrogen balance through the supply of exogenous carbon and nitrogen. The following steps outline the removal and culturing of the endosperm.

[0050] 1) Select rice seeds where both the embryo and the endosperm have emerged and are growing uniformly. Remove 2 / 3 of the endosperm (approximately 1 / 3 of the seed volume) along the lower edge of the embryo. Figure 4 A), or remove all the endosperm.

[0051] 2) Place the seeds with the endosperm removed into new 9 cm culture dishes. Add 8 mL of 1 / 2 Kimura B liquid medium with four carbon-nitrogen concentrations: C:N=0:0 (low carbon, low nitrogen), C:N=15 mM:2.5 mM (high carbon, high nitrogen), C:N=15 mM:0 (high carbon, low nitrogen), and C:N=0:2.5 mM (low carbon, high nitrogen) to different culture dishes. Seal the dishes with sealing film and place them in the tissue culture room for incubation (temperature: 25-28℃, light / dark = 12 h / 12 ​​h). This is day 0.

[0052] The results are as follows Figure 4 As shown, Figure 4 The growth of rice seeds with different proportions of endosperm removed two days after germination was compared in sterile ultrapure water. Figure 4 B, C, and D in the study statistically analyzed the stem and leaf length, root length, and crown root number of rice seedlings after 10 days of continuous growth. It was found that the growth of rice seedlings with 2 / 3 of the endosperm removed and those with all of the endosperm removed were significantly inhibited after the 7th day. However, removing 2 / 3 of the endosperm was easier to perform than removing all of the endosperm, was less likely to damage the embryo, and ensured normal seedling development. Therefore, it was determined that 2 / 3 of the endosperm should be removed for subsequent experiments.

[0053] 5. Change the fluid 1) Green embryos emerge on the second to third day. Select seedlings with uniform growth and place them in a 15 cm round petri dish. 2) Cover with the middle cover plate, add 40 mL of 1 / 2 Kimura B liquid culture medium with four different carbon and nitrogen concentrations, and completely submerge the rice. The culture medium should be gently shaken frequently to ensure even distribution of nutrients and to increase the oxygen content in the water to prevent waterlogging stress. Do this 3-4 times a day. 3) Phenotypic observation and physiological measurement were performed on the seventh day of growth.

[0054] 6. Phenotypic identification and physiological testing 1) Stem and leaf length measurement: Rice seedlings were placed flat in 15 cm round dishes, photographed, and the stem and leaf length of the seedlings was counted using ImageJ software; 2) Measurement of crown root number and total root length: All crown roots and taproots of rice seedlings were separated with tweezers, the number of crown roots was counted, the images were taken, and the total root length was counted using ImageJ; 3) Chlorophyll determination: Take 30-60 mg of rice seedlings, cut them into pieces, add 2 mL of extraction solution of acetone:ethanol:ultrapure water = 4.5:4.5:1, extract overnight at 4℃ in the dark for 12 h until the leaves turn completely white, measure the absorbance at wavelengths of 645 nm and 663 nm, and calculate the chlorophyll content.

[0055] 4) Determination of soluble sugar content: Take 50-100 mg of fresh sample, freeze it in liquid nitrogen, grind it into powder, add 1 mL of 10% methanol, vortex mix, sonicate at room temperature for 20 min, centrifuge at 12000 rpm for 10 min, take the supernatant for liquid chromatography analysis, and determine the content of glucose, fructose and sucrose.

[0056] like Figure 5 As shown, Figure 5 B, C, and D in the figure represent the statistics of stem and leaf length, crown root number, and root length of rice seedlings under different carbon and nitrogen concentrations, respectively. It was found that the number of crown roots of rice seedlings showed a significant response to high carbon and low nitrogen conditions, and the glucose concentration was finally set at 15 mM. Therefore, four different carbon and nitrogen concentration ratios were set: low carbon and low nitrogen (C:N=0:0, 0C0N), high carbon and high nitrogen (C:N=15 mM:2.5 mM, 15C2.5N), high carbon and low nitrogen (C:N=15 mM:0, 15C0N), and low carbon and high nitrogen (C:N=0:2.5 mM, 0C2.5N).

[0057] like Figure 6 As shown, after 7 days of growth under different carbon-nitrogen ratios, Nipponbare seedlings exhibited significant differences in stem and leaf length, crown root number, total root length, and chlorophyll content. Compared to low-carbon / low-nitrogen and high-nitrogen / high-carbon conditions, under high-carbon / low-carbon conditions, the number of crown roots and total root length of Nipponbare seedlings significantly increased, while chlorophyll content significantly decreased, and soluble sugars such as glucose, fructose, and sucrose accumulated significantly in the stems, leaves, and roots. Conversely, under low-carbon / high-nitrogen conditions, the number of crown roots and total root length significantly decreased, while the total stem and leaf length and chlorophyll content significantly increased, and the content of soluble sugars such as glucose, fructose, and sucrose in the stems, leaves, and roots significantly decreased.

[0058] like Figure 7 As shown, using the system of this study, Nipponbare seedlings were grown under a carbon-nitrogen imbalance condition of high carbon and low nitrogen for 3 days, and then transferred to a balanced carbon-nitrogen condition of low carbon and low nitrogen and high carbon and high nitrogen for 4 days. It was observed that the number of crown roots and the total root length of Nipponbare seedlings decreased significantly, while the chlorophyll content increased significantly. Similarly, when seedlings were placed under the same treatment of low carbon and high nitrogen, the number of crown roots and the total root length increased significantly.

[0059] 7. Results Analysis All of these phenomena demonstrate that different carbon-nitrogen ratios can regulate the growth and development of rice seedlings. The device system of this invention can be used to study the regulation of crop seedling growth by carbon-nitrogen ratios.

[0060] 1) The stem and leaf lengths of low-carbon / low-nitrogen and high-carbon / low-nitrogen seedlings were significantly shorter than those of high-carbon / high-nitrogen and low-carbon / high-nitrogen seedlings, indicating that stem and leaf length does not specifically respond to the carbon-nitrogen ratio; 2) Compared to low-carbon / low-nitrogen and high-carbon / high-nitrogen conditions, the number of crown roots, total root length, and soluble sugar content of rice seedlings significantly increased, while chlorophyll content significantly decreased under high-carbon / low-nitrogen conditions; whereas, the number of crown roots, total root length, and soluble sugar content of rice seedlings significantly decreased under low-carbon / high-nitrogen conditions. These results indicate that carbon-nitrogen balance can regulate the number of crown roots, total root length, chlorophyll content, and soluble sugar content of rice.

[0061] This invention uses rice as an example to construct the system. If other crops are used, the proportion of endosperm removal, the supply of glucose and total nitrogen, and the supply of various reagents and nutrient solutions need to be explored by oneself. However, this circular culture dish and intermediate cover plate device is applicable to most crops, or the size of the culture dish and intermediate cover plate can be adjusted according to the growth characteristics of different crops.

[0062] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.

Claims

1. A device for precisely controlling crop carbon nutrient supply and studying the interaction between carbon and mineral nutrients, characterized in that, The device includes a petri dish and an intermediate cover plate (2). The petri dish includes a dish body (3) and a dish lid (1). The intermediate cover plate (2) is provided with an upward support (21) and a downward leg (22). The leg (22) of the intermediate cover plate is supported inside the dish body (3) of the petri dish, and the support (21) of the intermediate cover plate supports the dish lid (1) of the petri dish upward.

2. The apparatus according to claim 1, characterized in that, The intermediate cover plate is provided with a plurality of holes (23), which are evenly or unevenly distributed on the intermediate cover plate.

3. The apparatus according to claim 3, characterized in that, The cross-sectional shape of the culture dish is circular, square, rectangular, or irregular. The height of the dish body is 5-100 mm, the height of the lid is 5-100 mm, and the area of ​​the dish body and lid is 2826-160000 mm². 2 .

4. The apparatus according to claim 1, characterized in that, The shape of the intermediate cover plate (2) is adapted to the shape of the petri dish. The intermediate cover plate (2) is placed inside the petri dish body (3), and the petri dish lid (1) is placed on the petri dish body (3). The intermediate cover plate (2) divides the inner space of the petri dish into an upper part and a lower part. The height of the lower part is the length of the legs (22). The thickness of the intermediate cover plate (2) is 0.3-3 mm. The legs (22) are vertically bent downward along the edge of the intermediate cover plate. There are 4-8 legs. The supports (21) are vertically bent upward along the edge of the intermediate cover plate. There are 2-8 supports. The length of the legs is 1100 mm and the width is 3-20 mm. The length of the supports is 5-100 mm and the width is 3-20 mm. The diameter of the holes is 1-10 mm and the pore density is 0.1-5 pores / cm. 2 .

5. The apparatus according to any one of claims 1-4, characterized in that, The culture dish is made of a highly transparent polymer material or transparent glass material, wherein the highly transparent polymer material is polycarbonate or polystyrene; the middle cover plate is made of a highly transparent polycarbonate or polystyrene material.

6. A method for studying the interaction between crop carbon nutrition and mineral nutrients, characterized in that, The method includes the step of immersing seeds with a portion of the endosperm removed in a culture medium below an intermediate cover plate using the apparatus described in any one of claims 1-4, wherein the culture medium is a nutrient solution containing different proportions of carbon and other mineral nutrients.

7. The method according to claim 5, characterized in that, The amount of culture medium used is selected based on the area of ​​the petri dish, the height of the middle cover stand, and the crop seedlings, so that the height of the culture medium in the petri dish just covers the entire seedling; the culture medium is changed every 2-4 days.

8. The method according to claim 5, characterized in that, The other nutrients are nitrogen or other mineral elements, including phosphorus, sulfur, potassium, calcium, magnesium, iron, zinc, manganese, boron, nickel, molybdenum, copper and chlorine; the culture is carried out under sterile conditions.

9. The apparatus according to any one of claims 1-5 or the method according to any one of claims 6-8, characterized in that, The crop is rice, wheat, corn, or soybean.

10. The method according to any one of claims 6-8, characterized in that, The crop is rice, and the endosperm is removed in a ratio of 2 / 3.

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