CN114680103B

CN114680103B - Method for preparing plant disease specimen

Info

Publication number: CN114680103B
Application number: CN202210399005.0A
Authority: CN
Inventors: 闫金姣; 徐记迪; 费昭雪; 贾彩霞; 陈亚菲; 李培琴
Original assignee: Northwest A&F University
Current assignee: Northwest A&F University
Priority date: 2022-04-15
Filing date: 2022-04-15
Publication date: 2023-03-14
Anticipated expiration: 2042-04-15
Also published as: CN114680103A

Abstract

The invention discloses a method for manufacturing a plant disease specimen, which belongs to the field of specimen manufacturing and comprises the following steps: (1) sampling: selecting a typical sample with obvious plant disease symptom symptoms according to needs; (2) primary drying: placing the sample in an oven for primary drying; (3) fixing treatment: spraying a color retention fixing agent on the surface of the sample after primary drying, and standing at normal temperature; (4) secondary drying: placing the fixed sample in an oven for secondary drying; (5) composite drying: embedding the sample after the secondary drying into silica gel particles, and completely drying in an oven; (6) shaping: and (3) spraying a color retention fixing agent on the surface of the completely dried sample, embedding the sample in epoxy resin AB glue after complete drying, and solidifying to obtain the plant disease specimen. So as to solve the problems of distortion of plant disease specimens, incomplete sample forms, decay and color change of the specimens.

Description

Method for preparing plant disease specimen

Technical Field

The invention relates to the field of specimen preparation, in particular to a method for preparing a plant disease specimen.

Background

The plant disease specimen is an important experimental material for relevant specialties such as forestry, plant protection, gardens, agriculture and gardening. At present, the specimen is mainly of the types of boxed specimen, dipped specimen, cured leaf specimen, slide specimen and the like. The slide specimen is suitable for observation under a microscope, has a mature manufacturing system and can be stored for a long time; the box-packed specimens and the cured leaf specimens are easy to fade along with the storage time, have unobvious symptoms of pests and diseases, and need to invest a great deal of energy to manage, thereby causing great inconvenience and trouble to the experiment teaching of the department. Therefore, the development of a novel plant disease specimen which is three-dimensional, visual, undistorted, easy to store and capable of being stored for a long time is of great significance.

The epoxy resin AB glue is also called crystal glue, is a novel material widely applied to amber specimens in recent years, is a two-component high polymer material consisting of epoxy resin and a curing agent, has the advantages of no toxicity, no environmental pollution, low cost and the like, and has a certain research basis in common plant specimens and Chinese herbal medicine specimens. However, the proportion of the AB glue, the solidification temperature, the plant type and the treatment method in the process of preparing the specimen all influence the final specimen, for example, the difference of the proportion of the A glue and the B glue can cause the specimen material to sink and not easily precipitate, the defoaming capability of the specimen material is influenced, the transparency is finally influenced, the problems of discoloration, slow solidification and bubble precipitation of the plant and the colloid can be caused by overhigh or overlow solidification temperature, and if the plant treatment is poorer, the phenomena of mildew, rot, discoloration and the like of the specimen can be caused. In addition, since the plant disease specimens need to clearly show typical symptoms and diseases, the plant disease specimens in different stages need to be displayed, such as powdery mildew, black spot, rust and other diseases which are common in plants, and the symptoms of the diseases are different in each disease stage, if the plants are not properly treated, not only the typical symptoms of the diseases are damaged, but also the plants are discolored, judgment is affected, such as powdery mildew drop and rust yellowish herpes discoloring, and therefore, the integrity of the plant symptoms and diseases needs to be ensured as much as possible in the process of preparing the specimens. Therefore, the development of a novel plant disease specimen which is three-dimensional, visual, undistorted, easy to store and capable of being stored for a long time is of great significance.

Disclosure of Invention

In view of the above, the present invention aims to provide a method for preparing a specimen of a plant disease, so as to solve the problems of distortion of the specimen of the plant disease, incomplete sample form, decay and discoloration of the specimen.

The invention solves the technical problems through the following technical means:

a method for preparing a plant disease specimen comprises the following steps:

(1) Sampling: selecting a typical sample with obvious plant disease symptom symptoms according to needs;

(2) Primary drying: placing the sample in an oven for primary drying;

(3) Fixing treatment: spraying a color retention fixing agent on the surface of the sample after primary drying, and standing at normal temperature;

(4) Secondary drying: placing the fixed sample in an oven for secondary drying;

(5) And (3) composite drying: embedding the sample after the secondary drying into silica gel particles, and completely drying in an oven;

(6) Shaping: and (3) spraying a color retention fixing agent on the surface of the completely dried sample, embedding the sample in epoxy resin AB glue after complete drying, and solidifying to obtain the plant disease specimen.

The plant is dried by adopting the oven, which is a method for removing the water in the plant body quickly, maintaining the original shape of the plant to be higher and degrading the color to be the lowest, but the long-time drying can cause the plant to be black or crisp, so that the whole disease judgment of the specimen is influenced, therefore, the invention adopts multiple times of drying to avoid the problems, adopts low-temperature slow drying for primary drying, and removes the water in the plant as much as possible while maintaining the original shape of the plant color. In addition, in the process of manufacturing the plant disease specimen, the powder spots and the powdery mildew layers on the surface of the specimen need to be kept from falling off or the color of the original disease spots is kept, so that the specimen is fixed by adopting a color retention fixing agent after primary drying, the powder spots are prevented from being browned in the process of secondary heating and drying, the thinner parts of the disease spots are prevented from being black and crisp, and finally, the drying is carried out by adopting a mode of combining silica gel particles and baking. Through primary drying and secondary drying, the water content in the plant body is greatly reduced, but in order to avoid the phenomenon that the silica gel compresses and deforms the speckles of the speckles, silica gel particles are combined with a drying technology, so that the using amount of the silica gel particles is reduced, the drying time is shortened, and the initial shape of the plant is maintained as much as possible.

Furthermore, the method disclosed by the invention is especially suitable for preparing specimens from plants with black spot, anthracnose, powdery mildew, red blight and rust disease.

Further, in the primary drying step, the drying temperature is 25-30 ℃, the drying time is 20-30min, in the secondary drying step, the drying temperature is 35-40 ℃, the drying time is 10-15min, in the composite drying step, the drying temperature is 40-45 ℃, and the drying time is 10-15min.

Further, the mass ratio of the glue A to the glue B in the epoxy resin AB glue is 3:1.

further, the color-retaining fixing agent comprises the following raw materials: polyvinyl alcohol, sodium alginate, glycerol, quince seeds, tween and copper sulfate.

The color-preserving fixative can be used for keeping the integrity of disease symptoms besides the primary color of the plant specimen. The color-retaining fixing agent can be sprayed on plant specimens to form a film, so that the plant disease is shaped, the anti-deformation capability of plants is improved, and the disease symptoms are kept intact. In order to avoid the problem that a film formed in the plant drying process is tilted or falls off, the adhesion between the color-retaining fixing agent and the plant needs to be increased, and meanwhile, the formed film does not influence the appearance of a plant specimen, and the sodium alginate used alone has high viscosity and is easy to fall off after being dried, so the sodium alginate needs to be compounded with other raw materials for modification.

Polyvinyl alcohol mixes with glycerine, tween and stirs into emulsion-like micromolecule together, both can evenly disperse in sodium alginate gel, also can reduce the viscidity of sodium alginate and improve the film forming property, later compound sodium alginate gel after will compounding and the viscidity concentrate that quince seed was decocted into is compound, obtain the hydrophilic oleophilic type viscidity liquid that can form the membrane after stirring evenly under the condition of heating, this viscidity liquid has higher heat stability simultaneously and prevents that the membrane from ftractureing in drying process, drop and the even spraying of higher flow property to the plant surface, also can guarantee simultaneously that the plant shape is fixed, prevent to encapsulate plant specimen shrinkage and discolour after epoxy. If the epoxy resin is damped or air or water vapor can be separated out under the low-temperature condition, the specimen sealed in the epoxy resin becomes damp and mildews, and the color-preserving fixing agent is used for isolating the contact of the plant specimen with the air and the water vapor after being formed into a film, so that the specimen sealed in the epoxy resin is prevented from mildewing and decaying.

Further, the mass ratio of the sodium alginate to the quince tree seeds is 1:8, the mass ratio of the sodium alginate to the polyvinyl alcohol is 1: (0.5-0.6).

Further, the mass ratio of the polyvinyl alcohol to the tween to the glycerol is 1:0.15:0.2, wherein the mass ratio of the copper sulfate to the polyvinyl alcohol is 0.05:1.

further, the preparation method of the color-retaining fixing agent comprises the following steps:

mixing sodium alginate with water to prepare gel, mixing polyvinyl alcohol, tween, glycerol and water, stirring until demulsification is performed, adding into the gel, and uniformly mixing to obtain composite gel;

decocting quince tree seeds with water for 1-2h, filtering tree seeds after decocting, heating and concentrating to obtain a concentrated solution;

mixing the concentrated solution, the composite gel and copper sulfate, heating to 30-40 deg.C, stirring for 20-30min, and standing for 10-12h to obtain color-retaining fixative.

Further, the color retention fixing agent is diluted by water for use, and the volume ratio of color retention fixing to water is 1: (0.5-1). When the disease of the prepared specimen is powdery mildew layer and other similar symptoms, the volume ratio of the color-retaining fixing agent to water is 1:0.5-0.6, when the diseases of the specimen are black spot and the like with spot-shaped symptoms, 1: a ratio of 0.8 to 1.

Further, in the setting step, after the sample is embedded in the epoxy resin AB glue, standing for 15-20min for defoaming in the environment of 30-40 ℃, and then solidifying for 24h at the temperature of 25-35 ℃.

Further, the method also comprises a step of manufacturing a label and/or a two-dimensional code label after the plant disease specimen is manufactured, wherein the label and/or the two-dimensional code label comprise a disease name, a host name, a pathogen academic name, a collection place and a manufacturer.

Has the advantages that:

1. the plant disease specimen is subjected to multiple times of low-temperature drying and composite drying, so that the water content in the specimen is remarkably reduced, meanwhile, the powder spots and the powdery mildew layers on the surface of the specimen are kept not to fall off or the color of the original disease spots is kept, and the color of the specimen does not fade after the specimen is placed for a long time.

2. The color-preserving fixing agent prepared from the raw materials of sodium alginate, polyvinyl alcohol, quince seeds and the like can not only keep the original color of a plant specimen, but also shape the plant disease, improve the anti-deformation capability of the plant and keep the disease symptoms intact.

3. The plant disease specimen prepared by the invention can maintain good quality for a long time, and does not have the phenomena of fading, shrinking, mildewing, rotting and the like.

Drawings

FIG. 1: the red blight specimen of osmanthus fragrans prepared in example 3;

FIG. 2: a specimen label and a two-dimensional code label schematic;

FIG. 3: white powdery mildew specimens of mulberry prepared in example 4.

Detailed Description

The invention will be described in detail below with reference to examples and the accompanying drawings:

example 1: preparation of color-retaining fixing agent

Before preparing a plant disease specimen, a color-retaining fixing agent needs to be prepared, the raw material proportion is shown in table 1, and the specific steps are as follows:

TABLE 1 (unit: g)

	Polyvinyl alcohol	Sodium alginate	Glycerol	Quince tree seeds	Tween-40	Copper sulfate
							Example 1	5	10	1	80	0.75	0.25
Example 2	6	10	1.2	80	0.9	0.3
							Comparative example 1	5	10	1	0	0.75	0.25
Comparative example 2	5	0	1	80	0.75	0.25
							Comparative example 3	5	10	0	80	0	0.25
Comparative example 4	5	10	1	80	0.75	0
							Comparative example 5	0	10	1	80	0.75	0.25
Comparative example 6	5	10	1	80	0.75	0.25

The preparation methods of the color retention fixing agents of the embodiment 1 and the embodiment 2 are as follows:

weighing the raw materials according to the mass in the table 1, mixing sodium alginate with 200g of water, uniformly stirring to prepare gel, mixing polyvinyl alcohol, tween, glycerol and water, stirring at 800rpm for 1h until demulsification is carried out, mixing with the gel, and uniformly mixing and stirring to obtain composite gel;

mixing the air-dried and crushed quince seeds with 1000g of water, decocting with strong fire for 1h, standing for 24h after the decoction is finished, filtering the seeds, and heating and concentrating to 200ml to obtain a concentrated solution;

mixing the concentrated solution, the composite gel and copper sulfate, heating to 30 ℃, keeping the temperature, stirring for 20min, and standing for 12h to obtain the color-retaining fixing agent.

The preparation method of comparative example 1 is as follows:

mixing copper sulfate and water to obtain saturated solution, mixing with the composite gel, heating to 30 deg.C, stirring for 20min, and standing for 12 hr to obtain color-retaining fixative.

Comparative example 2:

weighing the raw materials according to the mass in the table 1, mixing polyvinyl alcohol, tween, glycerol and water, and stirring at 800rpm for 1h until demulsification is performed for later use;

mixing the concentrated solution, the emulsion breaking solution and copper sulfate, heating to 30 ℃, keeping the temperature, stirring for 20min, and standing for 12h to obtain the color retention fixing agent.

Comparative example 3:

weighing the raw materials according to the mass in the table 1, mixing sodium alginate with 200g of water and polyvinyl alcohol, and uniformly stirring to prepare gel;

mixing the concentrated solution, gel and copper sulfate, heating to 30 deg.C, stirring for 20min, and standing for 12 hr to obtain color-retaining fixative.

Comparative example 4:

mixing the concentrated solution and the composite gel, heating to 30 ℃, keeping the temperature, stirring for 20min, and standing for 12h to obtain the color-retaining fixing agent.

Comparative example 5:

weighing the raw materials according to the mass in the table 1, mixing sodium alginate with 200g of water, uniformly stirring to prepare gel, mixing tween, glycerol and water, stirring at 800rpm for 1h until demulsification is carried out, mixing with the gel, and uniformly mixing and stirring to obtain composite gel;

mixing the concentrated solution, the composite gel and copper sulfate, heating to 30 ℃, keeping the temperature, stirring for 20min, and standing for 12h to obtain the color retention fixative.

Comparative example 6:

weighing the raw materials according to the mass in the table 1, mixing sodium alginate with 200g of water, polyvinyl alcohol, tween and glycerol, and uniformly stirring to prepare composite gel;

Example 3: preparation of osmanthus fragrans red blight specimen

(1) Sampling: collecting sweet osmanthus leaves in the disease period of red blight after dew drying in the morning in sunny days, and selecting representative leaves with simple disease spots, obvious disease symptoms, and middle and upper parts of plants;

(2) Primary drying: placing the cinnamon leaf sample in an oven, and carrying out primary drying at 25 ℃ for 30min;

(3) Fixing treatment: uniformly spraying water on the surface of the primarily dried cassia leaves according to the ratio of 1:1, the diluted color retention fixative of the embodiment 1 is used for wetting the whole leaf, spraying the leaves 20-30cm away from the leaf, and standing at normal temperature for waiting for the color retention fixative to be completely dried;

(4) Secondary drying: placing the fixed folium Cinnamomi in oven for secondary drying at 35 deg.C for 15min, and turning over during drying;

(5) And (3) composite drying: the particle size of the selected silica gel particle drying agent is not more than 0.04cm, the cassia leaves after secondary drying are horizontally placed on the surface of the silica gel particles, then the silica gel particles are gently poured on the surface of the cassia leaves, the cassia leaves are completely embedded, the cassia leaves and the silica gel particles are simultaneously placed in a drying oven, the drying is carried out for 15min under the condition of 40 ℃, and the cassia leaves are completely dried;

(6) Shaping: uniformly spraying water on the surface of the completely dried sample according to the ratio of 1: the volume ratio of 1 is diluted, and the color-retaining fixing agent of the embodiment 1 is used for wetting the whole leaf, standing and air-drying. According to the following steps of 3:1, heating the A glue and the B glue to 30 ℃, standing for 20min, and defoaming. And (3) after the glue A and the glue B are uniformly mixed and stirred, slowly pouring epoxy resin into a mould, then pouring the epoxy resin into the mould, pouring the epoxy resin to the required height, if bubbles appear, removing the bubbles by using a needle, and then solidifying for 24 hours at the temperature of 25 ℃ to obtain the osmanthus fragrans red blight specimen, wherein the formula is shown in figure 1.

(7) Labeling: the method also comprises a label and a two-dimension code label manufacturing step after the plant disease specimen is manufactured, wherein the label content comprises a disease name, a host name, a pathogen academic name, a collection place and a manufacturer. Files, network addresses, multimedia data and the like suitable for the application of the two-dimensional code technology are inquired through encyclopedic, a relatively comprehensive plant disease specimen database is built, and the files and the network address information are made into two-dimensional codes of specimens by a two-dimensional code generator, as shown in fig. 2. The label or two-dimensional code label is then affixed to the specimen.

Example 4: preparation of white powdery mildew specimen of mulberry

(1) Sampling: collecting mulberry leaves in the disease period of powdery mildew in sunny morning after dew drying, and selecting representative leaves with simple disease spots, obvious disease symptoms and middle-upper parts of plants;

(2) Primary drying: placing the mulberry leaf sample in an oven, and primarily drying at 30 ℃ for 20min;

(3) Fixing treatment: uniformly spraying water on the surface of the primarily dried mulberry leaves, wherein the water is mixed with the mulberry leaves according to the weight ratio of 1: the diluted color retention fixative of the embodiment 2 with the volume ratio of 0.5 can be used for wetting the whole leaf, spraying the leaves 20-30cm away from the leaf, standing at normal temperature and waiting for the color retention fixative to be completely dried;

(4) Secondary drying: placing the fixed mulberry leaves in an oven for secondary drying at 40 deg.C for 10min, and turning over during drying;

(5) And (3) composite drying: the particle size of the selected silica gel particle drying agent is not more than 0.04cm, the mulberry leaves after secondary drying are placed on the surface of the silica gel particles, then the silica gel particles are poured onto the surface of the mulberry leaves gently, the mulberry leaves are completely embedded, the mulberry leaves and the silica gel particles are placed in a drying oven at the same time, the drying is carried out for 10min at the temperature of 45 ℃, and the mulberry leaves are completely dried;

(6) Shaping: and (3) uniformly spraying the color retention fixing agent of the embodiment 1 on the surface of the completely dried sample to completely wet the whole leaf, standing and air-drying. According to the following steps of 3:1, heating the A glue to 40 ℃, standing for 15min, and defoaming. And (3) after the glue A and the glue B are uniformly mixed and stirred, slowly pouring epoxy resin into a mould, then pouring the epoxy resin into the mould, pouring the epoxy resin to the required height, if bubbles appear, removing the bubbles by using a needle, and then solidifying for 24 hours at 35 ℃ to obtain the mulberry powdery mildew specimen, wherein the formula is shown in figure 3.

Example 5: the color-preserving fixing agent prepared in comparative examples 1-6 is used for preparing osmanthus fragrans red blight specimens, the specific preparation method is the same as that of example 3, and the specimens prepared in comparative examples 1-6 are compared with the specimens in example 3.

Comparative example 7:

the specimen preparation steps (1) to (2) were the same as in example 3;

(3) Secondary drying: placing the fixed folium Cinnamomi in oven for secondary drying at 35 deg.C for 15min, and turning over during drying;

(4) And (3) composite drying: the particle size of the selected silica gel particle drying agent is not more than 0.04cm, the cassia leaves after secondary drying are horizontally placed on the surface of the silica gel particles, then the silica gel particles are gently poured on the surface of the cassia leaves, the cassia leaves are completely embedded, the cassia leaves and the silica gel particles are simultaneously placed in a drying oven, the drying is carried out for 15min under the condition of 40 ℃, and the cassia leaves are completely dried;

(6) Shaping: coating a layer of completely dried bay leaves with epoxy resin UV adhesive for shaping treatment, and after complete solidification, coating the base material according to the ratio of 3:1, heating the A glue and the B glue to 30 ℃, standing for 20min, and defoaming. And (3) slowly pouring epoxy resin into the mold after the glue A and the glue B are uniformly mixed and stirred, pouring the epoxy resin to the required height after the specimen is placed into the mold, driving out the bubbles by using a needle if the bubbles appear, and then solidifying for 24 hours at the temperature of 25 ℃ to obtain the red blight specimen of the osmanthus fragrans.

Comparative example 8:

(1) Sampling: collecting sweet osmanthus leaves in the disease stage of red blight after dew drying in the morning of sunny days, and selecting representative leaves with simple disease spots, obvious disease symptoms, and middle-upper parts of plants;

(2) And (3) drying silica gel: the particle size of the selected silica gel particle desiccant is not more than 0.04cm, the bay leaves are flatly placed on the surfaces of the silica gel particles, then the silica gel particles are gently poured on the surfaces of the bay leaves, and the bay leaves are completely embedded until the moisture in the bay leaves is completely sucked out;

(3) Shaping: according to the following steps of 3:1, heating the A glue and the B glue to 30 ℃, standing for 20min, and defoaming. And (3) slowly pouring epoxy resin into the mold after the glue A and the glue B are uniformly mixed and stirred, pouring the epoxy resin to the required height after the specimen is placed into the mold, expelling the bubbles by using a needle if the bubbles appear, and solidifying for 24 hours at the temperature of 25 ℃ to obtain the osmanthus fragrans red blight specimen.

Comparative example 9:

(2) And (3) drying: placing the cinnamon leaf sample in a drying oven, and drying for 60min at the temperature of 45 ℃;

(3) Shaping: according to the following steps of 3:1, heating the A glue and the B glue to 30 ℃, standing for 20min, and defoaming. And (3) slowly pouring epoxy resin into the mold after the glue A and the glue B are uniformly mixed and stirred, pouring the epoxy resin to the required height after the specimen is placed into the mold, driving out the bubbles by using a needle if the bubbles appear, and then solidifying for 24 hours at the temperature of 25 ℃ to obtain the red blight specimen of the osmanthus fragrans.

The specification and the size of all the specimens are as follows: length and width: 5X 8cm, thickness 2cm.

1. The specimens were observed indoors in a place protected from direct sunlight, the conditions of plants in the specimens were recorded every 3 months for specimens prepared in 2020.1, the observation time was 12 months in total, and the obtained data are shown in table 2:

wherein (-) represents: the color of the implant in the specimen is unchanged; no significant shrinkage occurred; the whole plant has no mildew and rot;

wherein (+) represents: the color of the plants in the specimen becomes yellow and faded; slightly shrinking; mildew stains appear;

wherein (++) denotes: the color of the plants in the specimen is yellow and the color is seriously faded; significant shrinkage; mildew stains are serious, and rottenness occurs;

TABLE 2

And (3) data analysis:

1. the specimen prepared in the example 3 has no phenomena of fading, yellowing and the like after 12 months, has clear disease spots, keeps the original color of the specimen for a long time, has no phenomenon of generating gaps in dripping glue due to volume shrinkage, and has no phenomenon of mildewing and rotting of the specimen.

2. Comparative examples 1 and 2 compared with example 1, the color-retaining fixative prepared without adding quince seeds and sodium alginate respectively has no obvious change on the whole of the specimen of example 3 in the first observation, but the specimens of comparative examples 1 and 2 begin to fade in the second observation, the discoloration is serious and mildewing occurs in the third observation, and the shrinkage occurs in comparative example 2. The color of the specimens is seriously faded and mildewed and rotten when the specimens are observed for the fourth time, the specimens are seriously shrunk in the comparative example 2, the humidity in the air is obviously improved before and after the observation for the second time in rainy season, and the mildewed and rotten phenomena of the specimens are caused. Therefore, sodium alginate or quince seeds alone cannot form a protective film on the surface of the specimen well, and the specimen is discolored, rotted and shriveled.

3. Comparative example 3 compared with example 3, no glycerol and tween were added, comparative example 5 compared with example 3, no polyvinyl alcohol was added, and comparative example 6 compared with example 3, no demulsification operation was adopted, and the samples all deteriorated to some extent after a long time, especially comparative example 5, and the phenomena of fading, shrinkage and mildew occurred earlier. The film forming property of the color retention fixing agent can be reduced without adopting polyvinyl alcohol, so that the specimen is easy to fall off or crust in the drying process, excessive bubbles can be generated during the preparation of the specimen, and the sealing performance is reduced, and the phenomena of mildew and fading can be caused. However, if the polyvinyl alcohol is used alone, the regularity of polyvinyl alcohol molecules is not easy to break, and sodium alginate is not easy to be mixed uniformly, so that before the polyvinyl alcohol is used, the polyvinyl alcohol and glycerol and tween are demulsified into micromolecules, and then the sodium alginate is modified.

4. Comparative example 4 compared with example 3, copper sulfate is not added, and copper sulfate has a certain protection effect on the original color of the plant, because the original color of the specimen can be kept by adding the copper sulfate into the color-keeping fixing agent.

5. Compared with the example 3, the comparative examples 7, 8 and 9 have the advantages that the color retention fixative is not used for preparing the specimen in the comparative example 7, the silica gel drying method is used for preparing the specimen in the comparative example 8, and the oven drying method is used for preparing the specimen in the comparative example 9. Although the specimen of comparative example 7 was completely prepared, the protective film layer was not used to protect the fixed specimen and block water vapor, and the discoloration, shrinkage and mold formation occurred during the long-term storage similar to those of comparative examples 8 and 9.

Although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the spirit and scope of the invention as defined in the appended claims. The techniques, shapes, and configurations not described in detail in the present invention are all known techniques.

Claims

1. The method for manufacturing the plant disease specimen is characterized by comprising the following steps:

(2) Primary drying: placing the sample in an oven for primary drying;

(4) Secondary drying: placing the fixed sample in an oven for secondary drying;

(6) Shaping: spraying a color retention fixing agent on the surface of the completely dried sample, embedding the sample in epoxy resin AB glue after complete drying, and obtaining a plant disease specimen after solidification;

in the primary drying step, the drying temperature is 25-30 ℃, the drying time is 20-30min, in the secondary drying step, the drying temperature is 35-40 ℃, the drying time is 10-15min, in the composite drying step, the drying temperature is 40-45 ℃, and the drying time is 10-15min;

the color-retaining fixing agent comprises the following raw materials: polyvinyl alcohol, sodium alginate, glycerol, quince seeds, tween and copper sulfate; the mass ratio of the sodium alginate to the quince tree seeds is 1:8, the mass ratio of the sodium alginate to the polyvinyl alcohol is 1: (0.5-0.6); the mass ratio of the polyvinyl alcohol to the tween to the glycerol is 1:0.15:0.2, wherein the mass ratio of the copper sulfate to the polyvinyl alcohol is 0.05:1; the preparation method of the color retention fixing agent comprises the following steps:

2. The method for preparing a specimen for a plant disease according to claim 1, wherein the mass ratio of the glue A to the glue B in the epoxy resin AB glue is 3:1.

3. the method for preparing a plant disease specimen according to claim 2, wherein the color-retaining fixing agent is diluted with water and used, and the volume ratio of the color-retaining fixing agent to water is 1: (0.5-1).

4. The method for preparing a specimen for a plant disease according to claim 1, wherein in the step of setting, the sample is embedded in epoxy resin AB glue, and then is kept stand for 15-20min at 30-40 ℃ for defoaming, and then is set for 24h at 25-35 ℃.

5. The method for preparing a plant disease specimen according to claim 1, further comprising a step of preparing a label and/or a two-dimensional code label after the preparation of the plant disease specimen is completed, wherein the content of the label and/or the two-dimensional code label comprises a disease name, a host name, a pathogenic science name, a collection place and a preparation person.