Leguminous root nodule form retention agent, preparation method and use method of leguminous root nodule form retention agent

Abstract

The invention belongs to the technical field of plant specimen manufacturing, and particularly relates to a leguminous root nodule form retention agent and a preparation and use method thereof. The first component contains a dehydrating agent, a filling agent and a regulator for osmotic regulation, the second component contains an antiseptic bacteriostatic agent and a surface retaining agent, and the first component and the second component are stored separately. The method can be used for making leguminous root system root nodule specimens, so that the shapes and colors of leguminous root nodules can be better kept in the specimen making and long-term storage process.

Description

Technical Field

[0001] The invention belongs to the technical field of plant specimen preparation, and in particular relates to a leguminous root nodule morphology retaining agent, and a preparation method and a use method thereof. Background Art

[0002] The collection of whole plants or parts of plants to preserve them for a longer period of time is called a herbarium. The preparation of herbarium is a key tool in botanical research and teaching, and is intended to prolong the shelf life of the plant by preserving the whole plant or its parts. This process not only allows these specimens to be observed and studied at any time, but also provides important information on species identification and classification by maintaining the original morphology and color of the plant.

[0003] The traditional method of specimen preparation usually includes three steps: collection and trimming, pressing and drying, and binding and labeling. Collection and trimming means collecting healthy, complete, representative fresh plants or parts thereof; in the pressing and drying stage, pressure is applied to plant samples using special paper with high water absorption to remove moisture; binding and labeling means arranging, binding, labeling and archiving the dried specimens. However, there are some limitations in the current preparation of plant specimens, such as short color retention time, shriveled morphology, and easy mold. Due to its biochemical complexity, leguminous nodules are more susceptible to dehydration, shriveling, oxidation, blackening, and mold during the preservation process. These problems not only affect the ornamental value and scientific research use of the specimens, but also lead to the loss of important biological information. The reason why leguminous nodules are difficult to maintain their original morphology is mainly related to their biochemical characteristics. The nodules contain a high amount of water, and the water is lost rapidly during the specimen preparation process, resulting in the collapse of the cell structure, which makes the nodules shriveled and deformed. In addition, the abundant iron and other biometals in leguminous nodules are easily oxidized when exposed to oxygen, causing the surface of the nodules to turn black. Moreover, the enzymes and other organic substances in the nodules will undergo spontaneous chemical reactions during specimen processing, accelerating discoloration and tissue degradation. Therefore, it is urgent to provide a specimen preparation and preservation method that can better maintain the shape and color of leguminous nodules during specimen preparation and long-term preservation. Summary of the invention

[0004] The purpose of the present invention is to provide a leguminous nodule morphology preserving agent, preparation method and use method thereof in view of the deficiencies of the prior art. The leguminous nodule morphology preserving agent can be used for preparing leguminous root nodule specimens, so that the leguminous nodules can better maintain their morphology and color during specimen preparation and long-term storage.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0006] A leguminous root nodule morphology retainer comprises a first component containing a dehydrating agent, a filler and a regulator for osmotic regulation and a second component containing an antiseptic and an antibacterial agent and a surface retainer. The first component and the second component are stored separately.

[0007] According to the above scheme, the first component is a formaldehyde solution containing a dehydrating agent, a filler and a regulator for osmotic regulation.

[0008] According to the above scheme, the concentration of the filler in the first component is: 300.0g / L-700.0g / L, the concentration of the regulator used for osmotic regulation is 10.0g / L-100.0g / L; the concentration of the dehydrating agent is 0.5%-2.0wt%.

[0009] According to the above scheme, the filler is one or two of polyethylene glycol 200-600, polysorbate or carboxymethyl cellulose.

[0010] According to the above scheme, the regulator used for osmotic regulation is polyvinyl pyrrolidone.

[0011] According to the above scheme, the dehydrating agent is magnesium sulfate or magnesium chloride.

[0012] According to the above scheme, the concentration of the formaldehyde solution is 15-25%.

[0013] According to the above scheme, the second component is an anhydrous ethanol solution of an antiseptic and antibacterial agent and a surface retaining agent.

[0014] According to the above scheme, the preservative and antibacterial agent is one or a mixture of potassium sorbate, glufosinate, imazalil, and bacil.

[0015] According to the above scheme, the surface retaining agent is one or a mixture of polypropylene glycol or polyoxyethylene lauryl ether.

[0016] According to the above scheme, the concentrations of the preservative and antibacterial agent and the surface retaining agent are 0.5g / L-2.5g / L and 2.0g / L-25.0g / L respectively.

[0017] A preparation method of a leguminous nodule morphology retaining agent, the steps of which are as follows:

[0018] (1) dissolving a filler and a regulator for osmotic regulation in a formaldehyde solution at concentrations of 300.0 g / L to 700.0 g / L and 10.0 g / L to 100.0 g / L, respectively, to obtain a solution A;

[0019] (2) dissolving the dehydrating agent in solution A at a concentration of 0.5% to 2.0% to obtain solution B, i.e., the first component;

[0020] (3) Dissolve the preservative and antibacterial agent and the surface retaining agent in anhydrous ethanol at concentrations of 0.5 g / L-2.5 g / L and 2.0 g / L-25.0 g / L, respectively, to obtain solution C, i.e., the second component, which is stored for later use.

[0021] The method for using the above-mentioned leguminous nodule morphology retaining agent comprises the following steps:

[0022] (1) soaking the leguminous root nodule sample in the first component for 5-10 days and then taking it out;

[0023] (2) Soak the leguminous root nodule sample in the second component for 10-15 minutes, then take it out and dry it under ventilation at room temperature;

[0024] (3) Preservation of leguminous root nodule samples.

[0025] According to the above scheme, before soaking, the leguminous root nodule sample is first soaked in a sodium hypochlorite solution for disinfection and then taken out. The concentration of the sodium hypochlorite solution is 20-30%, and the disinfection and sterilization time is 5-10 minutes.

[0026] According to the above scheme, the leguminous root nodule samples are immersed in the first component for 5-10 days.

[0027] According to the above scheme, the leguminous root nodule sample is immersed in the second component for 10-15 minutes.

[0028] According to the above scheme, the preservation in step (3) is to dry the leguminous root nodule sample for preservation, or to make it into a dripping sample for preservation, or to make it into a liquid sample for preservation.

[0029] The leguminous nodule morphology retaining agent provided by the present invention uses a first component containing a dehydrating agent, a filler and a regulator for osmotic regulation, and promotes the replacement of water in the leguminous nodules by the filler under the action of the dehydrating agent, thereby maintaining the original morphology of the leguminous nodules. At the same time, the added regulator has an osmotic regulation effect on water molecules inside and outside the nodules, which is beneficial to maintaining the morphology of the nodules and preventing the nodules from collapsing due to excessive water loss, thereby solving the problem of deformation and shriveling of sample nodules in specimen preparation.

[0030] Then, the second component containing preservatives and antibacterial agents and surface retainers is used to inhibit the growth of bacteria on the nodule surface, reduce nodule decay and discoloration, and form a stable protective film on the nodule surface, which synergistically improves the morphology and color preservation period of leguminous nodules, which is beneficial to the long-term preservation of leguminous root nodule specimens.

[0031] Beneficial effects of the present invention:

[0032] (1) The preparation and use methods of a leguminous nodule morphology retainer provided by the present invention can prevent the nodules from losing water and collapsing, and is conducive to the stable maintenance of the morphology of leguminous nodules, by infiltrating the dehydrating agent, filler and regulator inside and outside the nodules, and using the second component containing the preservative and antibacterial agent and the surface retainer. The preparation method can effectively maintain the integrity of the morphology and size of the root nodule specimens, so that the leguminous nodules can better maintain their morphology and color during the specimen preparation and long-term storage, and solve the technical problems of nodule deformation, shriveling, dehydration, shrinkage, decay and discoloration in specimen preparation.

[0033] (2) The method of the present invention is simple to operate and can be used for the morphological maintenance, processing and preparation of leguminous root nodule specimens. The plant can be directly soaked after washing, which is simple to operate and does not require the use of high temperature or freeze drying. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The soybean root nodule sample treated with the leguminous nodule morphology retaining agent of Example 1 ( Figure 1 b) and control samples (soybean root nodule samples not treated with the legume nodule morphology maintaining agent of Example 1, Figure 1 a) photos;

[0035] Figure 2 This is a photograph of a soybean root nodule liquid-immersed specimen treated with the leguminous nodule morphology preserving agent of Example 1;

[0036] Figure 3 This is a peanut root nodule glue encapsulation sample treated with the leguminous nodule morphology retaining agent of Example 2 ( Figure 3 b) and control samples (peanut root nodule glue-encapsulated samples not treated with the leguminous nodule morphology retaining agent of Example 2, Figure 3 a) Photographs. DETAILED DESCRIPTION

[0037] The present invention is further described below with specific implementation modes, but the present invention is not limited to these embodiments.

[0038] Embodiment 1:

[0039] A leguminous root nodule morphology retainer comprises a first component containing polyethylene glycol (PEG500), polysorbate and polyvinyl pyrrolidone and a second component containing glufosinate, polypropylene glycol and polyoxyethylene lauryl ether. The first component and the second component are stored separately.

[0040] Preparation method:

[0041] 1. Solution A: Mix 300 g of polyethylene glycol (PEG500), 300 g of polysorbate and 50 g of polyvinyl pyrrolidone in a 1000 mL beaker, add 1000 mL of 20% formaldehyde solution, stir thoroughly to dissolve, and obtain filling solution A.

[0042] 2. Solution B: Take 10g of magnesium sulfate and put it in a 1000mL beaker. Add 1000mL of solution A and stir thoroughly to dissolve to obtain solution B.

[0043] 3. Solution C: Take 1g of the preservative and antibacterial agent glufosinate, 5g of polypropylene glycol and 10g of polyoxyethylene lauryl ether in a 1000mL beaker, add 1000mL of anhydrous ethanol, stir thoroughly to dissolve to obtain Solution C.

[0044] application:

[0045] Soak the soybean root and nodule samples in 20% sodium hypochlorite solution, clean the surface and sterilize for 5 minutes before taking them out; soak the soybean root and nodule samples in solution B and take them out after 8 days; after ventilation and drying at room temperature, soak the soybean root and nodule samples in solution C for 15 minutes before taking them out and ventilation and drying at room temperature; finally, put the soybean root and nodule samples in a specimen frame for drying and preservation, see Figure 1 b.

[0046] Alternatively, soybean root and nodule samples can be preserved in liquid immersion specimens, see Figure 2 . Preserving specimens by immersion in liquid can better reflect the effects of morphology, color, etc.

[0047] Control samples: Soybean root nodule samples were not treated with the above-mentioned leguminous nodule morphology preserving agent, and the samples directly dried and stored, see Figure 1 a.

[0048] Soybean root nodule specimens ( Figure 1 a) The nodules become deformed, shrunken, dehydrated and blackened.

[0049] The soybean root nodule specimens prepared by treating the leguminous nodule morphology retaining agent of Example 1 ( Figure 1 b) The nodule morphology is full and can reflect the original morphology and color effects of the leguminous nodule morphology. The morphology and color of the soybean nodules can be stably maintained after long-term storage (at least half a year or more). This shows that the leguminous nodule morphology preserving agent provided by the present invention is used for the preparation of root nodule samples, which can prevent the nodules from losing water and collapsing, and is conducive to the stable maintenance of the morphology of soybean roots and nodules, solving the technical problems of nodule deformation, shriveling, dehydration, shrinkage, decay and discoloration in specimen preparation. The leguminous nodule morphology preserving agent treated with liquid can fully reflect the original morphology and color effects of the leguminous nodule morphology.

[0050] Embodiment 2:

[0051] A leguminous root nodule morphology retainer comprises a first component containing polyethylene glycol (PEG500), polysorbate and polyvinyl pyrrolidone and a second component containing potassium sorbate, polypropylene glycol and polyoxyethylene lauryl ether. The first component and the second component are stored separately.

[0052] Preparation method:

[0053] 1. Solution A: Mix 350 g of polyethylene glycol (PEG500), 350 g of polysorbate and 30 g of polyvinyl pyrrolidone in a 1000 mL beaker, add 1000 mL of 20% formaldehyde solution, stir thoroughly to dissolve, and obtain filling solution A.

[0054] 2. Solution B: Take 15g of magnesium sulfate and put it in a 1000mL beaker. Add 1000mL of solution A and stir thoroughly to dissolve to obtain solution B.

[0055] 3. Solution C: Take 2g of preservative and antibacterial agent potassium sorbate, 10g of polypropylene glycol and 15g of polyoxyethylene lauryl ether in a 1000mL beaker, add 1000mL of anhydrous ethanol, stir thoroughly to dissolve to obtain Solution C.

[0056] application:

[0057] Soak the peanut root nodule samples in 20% sodium hypochlorite solution, clean the surface and disinfect for 10 minutes before taking them out; after ventilation and drying at room temperature, soak the soybean root and nodule samples in solution B and take them out after 6 days; after ventilation and drying at room temperature, soak the peanut root and nodule samples in solution C for 8 minutes before taking them out and ventilation and drying at room temperature; finally, encapsulate the peanut root nodule samples with glue to maintain the morphology of the peanut nodule specimens. Figure 3 b.

[0058] Control sample: The peanut root nodule sample was not treated with the above-mentioned leguminous nodule morphology retaining agent and was directly encapsulated with glue. Figure 3 a.

[0059] By comparison, we can see that:

[0060] Peanut root nodule glue-encapsulated specimens ( Figure 3 a) The root system loses color, the nodules become deformed and shrunken, and the nodules turn black.

[0061] Peanut root nodule glue-encapsulated specimens prepared by treating the leguminous nodule morphology retaining agent of Example 2 ( Figure 3b) The nodules are full in shape, reflecting the original shape and color of the peanut nodules. The shape and color of the peanut nodules can be stably maintained after long-term storage (at least two years). It can prevent the nodules from losing water and collapsing, and is conducive to the stable maintenance of the shape of the peanut roots and nodules, solving the technical problems of deformation, shriveling, dehydration, shrinkage, decay and discoloration of the nodules in specimen preparation.

[0062] It should be understood that those skilled in the art can make improvements or changes based on the above description, and all these improvements and changes should fall within the scope of protection of the appended claims of the present invention.

Claims

1. A leguminous nodule morphology retaining agent, characterized in that: The invention comprises a first component containing a dehydrating agent, a filler and a regulator for osmotic regulation and a second component containing an antiseptic and an antibacterial agent and a surface retaining agent, wherein the first component and the second component are stored separately.

2. The leguminous nodule morphology retaining agent according to claim 1, characterized in that: The first component is a formaldehyde solution containing a dehydrating agent, a filler and a regulator for osmotic regulation; the second component is an anhydrous ethanol solution containing an antiseptic and antibacterial agent and a surface retaining agent.

3. The leguminous nodule morphology retaining agent according to claim 1 or 2, characterized in that: The filler is one or a mixture of polyethylene glycol 200-600, polysorbate or carboxymethyl cellulose; the regulator for osmotic regulation is polyvinyl pyrrolidone; and the dehydrating agent is magnesium sulfate or magnesium chloride.

4. The leguminous nodule morphology retaining agent according to claim 1 or 2, characterized in that: The antiseptic and antibacterial agent is one or a mixture of potassium sorbate, glufosinate, imazalil or bacil; the surface retaining agent is one or a mixture of two of polypropylene glycol or polyoxyethylene lauryl ether.

5. The leguminous nodule morphology retaining agent according to claim 2, characterized in that: The concentration of the formaldehyde solution is 15-25%.

6. The leguminous nodule morphology retaining agent according to claim 2, characterized in that: The concentrations of filler and regulator in the first component are 300.0g / L-700.0g / L and 10.0g / L-100.0g / L respectively; the concentration of dehydrating agent is 0.5%-2.0wt%; the concentrations of preservative and antibacterial agent and surface retaining agent are 0.5g / L-2.5g / L and 2.0g / L-25.0g / L respectively.

7. The method for preparing a leguminous nodule morphology retaining agent according to claim 1, characterized in that: Here are the steps: (1) dissolving a filler and a regulator for osmotic regulation in a formaldehyde solution at concentrations of 300.0 g / L to 700.0 g / L and 10.0 g / L to 100.0 g / L, respectively, to obtain a solution A; (2) dissolving the dehydrating agent in solution A at a concentration of 0.5% to 2.0% to obtain solution B, i.e., the first component; (3) Dissolve the preservative and antibacterial agent and the surface retaining agent in anhydrous ethanol at concentrations of 0.5 g / L-2.5 g / L and 2.0 g / L-25.0 g / L, respectively, to obtain solution C, i.e., the second component, which is stored for later use.

8. The method for using the leguminous nodule morphology retaining agent according to claim 1, characterized in that: The following steps are involved: (1) soaking the leguminous root nodule sample in the first component for 5-10 days and then taking it out; (2) Soak the leguminous root nodule sample in the second component for 10-15 minutes, then take it out and dry it under ventilation at room temperature; (3) Preservation of leguminous root nodule samples.

9. The method of use according to claim 8, characterized in that: Before the soaking, the leguminous root nodule sample is first soaked in a sodium hypochlorite solution for disinfection and then taken out.

10. The method of use according to claim 8, characterized in that: The legume nodule sample is immersed in the first component for 5-10 days; the legume nodule sample is immersed in the second component for 10-15 minutes.

Images