Method for rapidly preparing Frankliniella occidentalis identification slide specimen

Abstract

The invention relates to an insect specimen manufacturing method, in particular to a method for rapidly manufacturing a Frankliniella occidentalis identification slide specimen, which comprises the following steps: putting Frankliniella occidentalis into a 5% potassium hydroxide or sodium hydroxide test tube, and carrying out water bath at 50 DEG C for 15 minutes; soaking for 10 minutes by using distilled water after water bath; sequentially soaking the Frankliniella occidentalis in 75% alcohol for 5 minutes, 85% alcohol for 3 minutes, 95% alcohol for 2 minutes and 100% alcohol for 2 minutes; finally, soaking the Frankliniella occidentalis in 100% alcohol for 1 minute, and dripping glue on a glass slide; larva moving, posture adjusting and sheet covering are carried out; and a label is pasted after the slide is manufactured. The slide manufacturing method provided by the invention is simple in manufacturing and short in time consumption, keeps the integrity of Frankliniella occidentalis bodies in the manufacturing process, and can be used for rapid identification of Frankliniella occidentalis.

Description

Technical Field

[0001] The invention relates to an insect specimen preparation method, in particular to a method for rapidly preparing a slide specimen for identification of western flower thrips. Background Art

[0002] Frankliniella occidentalis, also known as western flower thrips, is a common name for insects in the order Thysanoptera. It is a microscopic insect with a body length of only 0.5 to 2.0 mm. Most thrips are herbivorous insects that mainly harm the floral organs and tender parts of plants. While harming plants, they can also spread plant pollen. Some species can also prey on mites while directly feeding on plants and spreading plant viral diseases to cause indirect harm. There are more than 500 species of host plants in 66 families of western flower thrips, which mainly harm vegetable crops such as peppers, eggplants, cucumbers, lettuce, and tomatoes. Western flower thrips harm the leaves, flowers, and fruits of plants, and use rasp-sucking mouthparts to absorb nutrients from plant tissues and organs to meet their own growth and development needs. They cause serious damage to crops by feeding, laying eggs, and spreading viruses.

[0003] Given that western flower thrips are small, hidden, and highly harmful, there is an urgent need to find a method for rapid identification of western flower thrips so that effective prevention and control measures can be taken in a timely manner to limit them to a controllable range. To accurately identify the species of western flower thrips by morphological identification, it is necessary to make slides, and good slide specimens can provide detailed morphological characteristics. The existing method for making slide specimens for identifying western flower thrips takes only 3 hours for the xylene transparent fixation step, and the process conditions are complex during the production process. During the production process, the wings and limbs of western flower thrips are prone to breakage, which affects the morphology of the insect body and causes deviations in the identification results. In order to overcome the above technical problems, it is urgent to find a method for quickly making slide specimens for identification of western flower thrips, and at the same time improve the integrity of the western flower thrips insect body during the specimen production process. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a method for quickly preparing glass slide specimens for identification of western flower thrips, aiming to simplify the preparation steps of glass slide specimens for identification of western flower thrips through a large number of experimental studies, greatly shorten the preparation time of glass slide specimens for identification of western flower thrips, and at the same time reduce the damage rate of insect bodies during the preparation process through method optimization, thereby improving the integrity of insect bodies.

[0005] Specifically, the present invention provides a method for quickly preparing a slide specimen for identifying western flower thrips, characterized in that the method comprises the following steps:

[0006] Step 1: Place the western flower thrips in an alkali solution for water bath; soak in distilled water after the water bath to allow the contents of the western flower thrips to be discharged;

[0007] Step 2, placing the western flower thrips treated in step 1 into alcohol solution and soaking them step by step to dehydrate them;

[0008] Step 3: drop neutral gum on a glass slide, place western flower thrips on the neutral gum, arrange the body parts of the western flower thrips, cover with a cover glass, attach a label, and prepare a slide specimen for identification of western flower thrips.

[0009] Furthermore, in step 1 of the method for rapidly preparing a slide specimen for identifying western flower thrips, the alkali solution is potassium hydroxide or sodium hydroxide.

[0010] Furthermore, in step 1 of the method for rapidly preparing a slide specimen for identification of western flower thrips, the alkali solution is 5% potassium hydroxide or 5% sodium hydroxide.

[0011] Furthermore, in step 1 of the method for rapidly preparing a slide specimen for identification of western flower thrips, the water bath is a 50° C. water bath for 15 minutes.

[0012] Furthermore, in step 1 of the method for rapidly preparing a slide specimen for identifying western flower thrips, the distilled water immersion time is 10 minutes.

[0013] Furthermore, in step 2 of the method for rapidly preparing a slide specimen for identifying western flower thrips, the concentration of the alcohol solution is 75%-100%.

[0014] Furthermore, in step 2 of the method for rapidly preparing slide specimens for identifying western flower thrips, the concentrations of the alcohol solution are 75%, 85%, 95%, and 100% respectively.

[0015] Furthermore, in step 2 of the method for rapidly preparing a slide specimen for identification of western flower thrips, the dehydration is performed by soaking the western flower thrips treated in step 1 in 75% alcohol for 5 minutes, then in 85% alcohol for 3 minutes, then in 95% alcohol for 2 minutes, then in 100% alcohol for 2 minutes, and finally in 100% alcohol for another 1 minute, using an alcohol concentration range of 75%-100%.

[0016] In this step, the alcohol concentration is gradually increased from 75% to 100%. Soaking the western flower thrips in different alcohol solutions can gradually dehydrate the western flower thrips body, so as not to cause the insect body to be too hard and brittle, resulting in the phenomenon of the insect body breaking during the specimen preparation process.

[0017] Furthermore, in step three of the method for quickly preparing a glass slide specimen for identification of western flower thrips, neutral gum is dropped on a glass slide, the western flower thrips is placed on the neutral gum, and under a dissecting microscope, various parts of the body of the western flower thrips are prodded with an insect pin, covered with a coverslip, and labeled after natural drying to obtain a glass slide specimen for identification of western flower thrips.

[0018] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages:

[0019] (1) The method for quickly preparing a slide specimen for identification of western flower thrips provided by the present invention shortens the time for the contents of the western flower thrips to be discharged to 25 minutes. In the prior art, the time for the contents of the western flower thrips to be discharged is as long as more than 2 hours. Compared with the prior art, the present invention greatly saves the specimen preparation time.

[0020] (2) The method for rapidly preparing glass slide specimens for identification of western flower thrips provided by the present invention omits the xylene transparent fixation step required for glass slide specimens of western flower thrips in the prior art. The xylene transparent fixation step alone takes up to 3 hours. Compared with the prior art, the present invention omits the xylene transparent fixation step through method optimization, thereby greatly shortening the specimen preparation time.

[0021] (3) In the method for rapidly preparing slide specimens for identification of western flower thrips provided by the present invention, the dehydration treatment is performed by soaking in 75% alcohol for 5 minutes, then soaking in 85% alcohol for 3 minutes, then soaking in 95% alcohol for 2 minutes, then soaking in 100% alcohol for 2 minutes, and finally soaking the western flower thrips in 100% alcohol for another 1 minute. The initial concentration of dehydration of western flower thrips is higher than that of the traditional method, and the dehydration time is shortened. This improvement not only shortens the preparation time of the present invention, but also keeps a certain toughness of the western flower thrips after dehydration. Compared with the traditional method, the present method is less likely to cause insect body damage during the specimen preparation process. DETAILED DESCRIPTION

[0022] The technical solution of the present invention is described below in conjunction with embodiments; however, the present invention is not limited to the following embodiments.

[0023] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below in conjunction with specific embodiments, but the embodiments are not intended to limit the present invention.

[0024] The experimental methods and detection methods described in the following embodiments are conventional methods unless otherwise specified; the reagents and materials are commercially available unless otherwise specified.

[0025] Example 1

[0026] This embodiment is for quickly preparing a slide specimen for identification of western flower thrips.

[0027] Neutral gum: purchased from Shanghai specimen model, China, refractive index: 1.5316.

[0028] Source of western flower thrips: In this example, western flower thrips were collected from potted tomatoes in the experimental shed of the Institute of Plant Protection, Ningxia Academy of Agriculture and Forestry Sciences.

[0029] The method for rapidly preparing slide specimens for identification of western flower thrips of the present invention is used to prepare 50 slide specimens for identification of western flower thrips. The specific steps are as follows:

[0030] Step 1: Place 50 western flower thrips in a test tube containing 5% potassium hydroxide or sodium hydroxide, and bathe them in a water bath at 50° C. for 15 minutes; after the water bath, deworm the western flower thrips, place them in a test tube containing distilled water and soak them for 10 minutes to discharge the contents in the body of the western flower thrips.

[0031] Step 2, soak the 50 western flower thrips treated in step 1 in 75% alcohol for 5 minutes, take them out and soak them in 85% alcohol for 3 minutes, take them out and soak them in 95% alcohol for 2 minutes, take them out and soak them in 100% alcohol for 2 minutes; finally, soak the western flower thrips in 100% alcohol for another 1 minute for standby use.

[0032] Step 3: Drop neutral gum on 50 slides, drop a drop of neutral gum in the center of each slide, place a western flower thrips on the glue, use an insect pin to poke the body parts of the western flower thrips so that the insect body is vertically located in the center of the slide, and after the insect body is completely placed, slowly lower the upper cover glass at an angle (no bubbles will be generated in the glue), dry naturally and then label, and obtain 50 western flower thrips identification slide specimens.

[0033] Example 2

[0034] This embodiment is for quickly preparing a slide specimen for identification of western flower thrips.

[0035] Neutral gum: purchased from Shanghai specimen model, China, refractive index: 1.5316.

[0036] Source of thrips: In this example, western flower thrips were collected from potted cucumbers in the experimental shed of the Institute of Plant Protection, Ningxia Academy of Agriculture and Forestry Sciences.

[0037] The method for rapidly preparing slide specimens for identification of western flower thrips of the present invention is used to prepare 50 slide specimens for identification of western flower thrips. The specific steps are as follows:

[0038] Step 1: Place 50 western flower thrips in a test tube containing 5% potassium hydroxide or sodium hydroxide, and bathe them in a water bath at 50° C. for 15 minutes; after the water bath, deworm the western flower thrips, place them in a test tube containing distilled water and soak them for 10 minutes to discharge the contents in the body of the western flower thrips.

[0039] Step 2, soak the 50 western flower thrips treated in step 1 in 75% alcohol for 5 minutes, take them out and soak them in 85% alcohol for 3 minutes, take them out and soak them in 95% alcohol for 2 minutes, take them out and soak them in 100% alcohol for 2 minutes; finally, soak the western flower thrips in 100% alcohol for another 1 minute for standby use.

[0040] Step 3: Drop neutral gum on 50 slides, drop a drop of neutral gum in the center of each slide, place a western flower thrips on the glue, use an insect pin to poke the body parts of the western flower thrips so that the insect body is vertically located in the center of the slide, and after the insect body is completely placed, slowly lower the upper cover glass at an angle (no bubbles will be generated in the glue), dry naturally and then label, and obtain 50 western flower thrips identification slide specimens.

[0041] Comparative Example 1

[0042] The comparative example is to prepare the slide specimen for identification of western flower thrips by the traditional method.

[0043] Source of western flower thrips: The comparative western flower thrips were collected from potted cucumbers in the experimental shed of the Institute of Plant Protection, Ningxia Academy of Agricultural and Forestry Sciences.

[0044] Prepare 50 slide specimens for identification of western flower thrips according to the following steps.

[0045] Step 1: Pick out about 50 western flower thrips from the alcohol solution and directly put them into a 6% NaOH solution at room temperature for 8 to 24 hours; the soaking time depends on the size and color of the insect body. The soaking time for large and dark-colored specimens is longer; a few specimens with very light colors can be directly dehydrated without decolorization;

[0046] Step 2: Take out the western flower thrips from the alkali solution, place it in distilled water for 2 hours to allow the body contents to be discharged naturally, and then dehydrate it step by step with alcohol solutions of different concentrations, that is, soak it in 30%, 60%, 85%, and 95% alcohol solutions for 5 to 10 minutes respectively, and then transfer the specimen to 100% alcohol and place it for 30 minutes;

[0047] Step 3: Use a straw to suck out the western flower thrips and spit them out on ordinary filter paper. Let the alcohol on the surface of the insect body dry naturally. Don't take too long, and make sure there is no "clear water" on the filter paper. Otherwise, there will be bubbles in the insect body of the glass slide. Then use an insect pin to gently poke the insect into xylene and soak it for 30 minutes.

[0048] Step 4: Put a drop of Canada gum in the center of the slide, place one end of the thrips on the glue, and under the dissecting microscope, use an insect pin to carefully pick out various parts of the insect body, and gently cover it with a coverslip. After the slide is prepared, place it on a flat plate, dry it naturally, and label it. This is how 50 slide specimens of western flower thrips identification are prepared.

[0049] Example 3

[0050] This example is to statistically analyze the insect body damage rates of the slide specimens prepared by the method of the present invention and the slide specimens prepared by the comparative example.

[0051] The breakage rate of each of the 50 glass slide specimens prepared in Example 1 and Example 2 was calculated, and the standard was that the body of the western flower thrips on the glass slide specimens was intact. All cases where the wings of the western flower thrips were broken, the antennae of the western flower thrips were missing, and the limbs were incomplete in the glass slide specimens for identification of the western flower thrips were counted as damaged. The breakage rate of each of the 50 glass slide specimens prepared in the comparative example was calculated, and the statistical standard was the same as that in Example 1 and Example 2, and the standard was that the body of the western flower thrips on the glass slide specimens was intact. All cases where the wings of the western flower thrips were broken, the antennae of the western flower thrips were missing, and the limbs were incomplete in the glass slide specimens for identification of the western flower thrips were counted as damaged. The breakage rate was calculated according to the following formula:

[0052] Damage rate (%) = (number of slide specimens containing damaged western flower thrips / total number of slide specimens) × 100

[0053] The statistical results of the damage rate of western flower thrips body in the slide specimens of western flower thrips identification prepared by different methods are shown in Table 1 below:

[0054] Table 1 Statistical results of the damage rate of western flower thrips in slide specimens prepared by different methods

[0055] Grouping Total number of specimens Number of complete specimens Number of damaged specimens Insect body damage rate (%) Example 1 50 50 0 0 Example 2 50 50 0 0 Comparative Example 50 38 12 24

[0056] As shown in Table 1, by using the method for rapidly preparing glass slide specimens for identification of western flower thrips provided by the present invention, during the preparation process, both Example 1 and Example 2 kept the western flower thrips body intact, while during the preparation process of the glass slide specimens for identification of western flower thrips by the comparative example, 3 cases of broken antennae of western flower thrips, 4 cases of broken wings of western flower thrips, and 5 cases of missing legs of western flower thrips occurred. A total of 12 cases of damaged insect body specimens appeared in the glass slide specimens of western flower thrips prepared by the proportional method, and the insect body damage rate during the preparation process was 24%, indicating that the method for rapidly preparing glass slide specimens for identification of western flower thrips provided by the present invention is far superior to the prior art in terms of insect body integrity, and the method provided by the present invention has more advantages in the rapid identification of western flower thrips.

[0057] In summary, the results show that compared with the prior art, the method for quickly preparing slide specimens for identification of western flower thrips provided by the present invention has simple preparation steps, greatly shortened preparation time, and the western flower thrips body remains intact during the preparation process, and can be used for rapid identification of western flower thrips.

[0058] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for rapidly preparing slide specimens for identification of western flower thrips, characterized in that: The method comprises the following steps: Step 1: Place the western flower thrips in an alkali solution for water bath; soak in distilled water after the water bath to allow the contents of the western flower thrips to be discharged; Step 2, placing the western flower thrips treated in step 1 into alcohol solution and soaking them step by step to dehydrate them; Step 3: Add neutral gum on a glass slide, place western flower thrips on the neutral gum, arrange the body parts of the western flower thrips, cover with a cover glass, and label to obtain a slide specimen for identification of western flower thrips.

2. The method for rapidly preparing slide specimens for identification of western flower thrips according to claim 1, characterized in that: The alkali solution in step 1 is potassium hydroxide solution or sodium hydroxide solution.

3. The method for rapidly preparing slide specimens for identification of western flower thrips according to claim 2, characterized in that: The alkali solution in step 1 is 5% potassium hydroxide solution or 5% sodium hydroxide solution.

4. The method for rapidly preparing slide specimens for identification of western flower thrips according to claim 1, characterized in that: The water bath in step 1 is 50° C. for 15 min.

5. The method for rapidly preparing slide specimens for identification of western flower thrips according to claim 1, characterized in that: The distilled water soaking time in step 1 is 10 minutes.

6. The method for rapidly preparing slide specimens for identification of western flower thrips according to claim 1, characterized in that: The concentration of the alcohol solution in step 2 is 75%-100%.

7. The method for rapidly preparing slide specimens for identification of western flower thrips according to claim 1, characterized in that: The concentrations of the alcohol solutions in step 2 are 75%, 85%, 95% and 100% respectively.

8. The method for rapidly preparing slide specimens for identification of western flower thrips according to claim 1, characterized in that: The dehydration in step 2 is to soak the western flower thrips treated in step 1 in 75% alcohol for 5 minutes, then soak it in 85% alcohol for 3 minutes, then soak it in 95% alcohol for 2 minutes, then soak it in 100% alcohol for 2 minutes, and finally soak the western flower thrips in 100% alcohol for another 1 minute.

9. The method for rapidly preparing slide specimens for identification of western flower thrips according to claim 1, characterized in that: In step three, use an insect pin to poke various parts of the body of the western flower thrips, cover them with a coverslip, dry them naturally, and then stick a label to prepare a slide specimen for identification of the western flower thrips.