Plant active oxygen rapid detection kit and application thereof

Abstract

The invention discloses a plant active oxygen rapid detection kit, which comprises a # imgabs0 # staining solution, an H2O2 staining solution and a decoloring solution, the # imgabs1 # staining solution is 0.5 mol / ml nitro tetrazole blue chloride, the H2O2 staining solution is 0.1 mol / ml diaminobenzidine, the decoloring solution is a mixed solution prepared by mixing absolute ethyl alcohol and glacial acetic acid according to a mass ratio of 3: 1, the mass ratio of the absolute ethyl alcohol to the glacial acetic acid is 1: 1, and the mass ratio of the absolute ethyl alcohol to the glacial acetic acid is 1: 1. The application of the plant active oxygen rapid detection kit in detection of plant # imgabs2 and H2O2 comprises the following steps: step 1, dyeing treatment, step 2, decoloration treatment, and step 3, observation and photographing. The kit can rapidly detect plant active oxygen, the process is rapid and time-saving, the operation is simple, detection can be performed only by using a water bath kettle and a microscope in the whole process, the cost is low, real-time distribution of # imgabs3 and H2O2 in plant roots and leaves can be seen, and meanwhile, the kit can be widely applied to herbaceous plants and woody plants and is wide in application range.

Description

Technical Field

[0001] The present invention relates to the field of life science technology, and in particular to a rapid detection kit for plant active oxygen and application thereof. Background Art

[0002] Existing test kits are mainly used to detect antioxidant enzymes, but reactive oxygen species themselves oxidize quickly, so timely detection is very necessary. However, there are no such test kits on the market.

[0003] Although we can prepare various drugs for determination, the various preparation methods are very complicated and poorly standardized. Moreover, there are certain disadvantages in detecting antioxidant enzymes in plant samples, and it is impossible to directly determine the The specific distribution of active oxygen in plant samples cannot be directly seen through visualization of H2O2. Therefore, the present invention proposes a plant active oxygen rapid detection kit and its application to solve the problems existing in the prior art. Summary of the invention

[0004] In view of the above problems, the purpose of the present invention is to provide a plant active oxygen rapid detection kit and its application, which can quickly detect plant active oxygen, the process is fast and time-saving, the operation is simple, the whole process only needs to use a water bath and a microscope for detection, the cost is low, and it can see And the real-time distribution of H2O2 in the roots and leaves of plants. At the same time, the kit can be widely used in herbaceous plants and woody plants, with a wide range of applications.

[0005] To achieve the purpose of the present invention, the present invention is implemented by the following technical scheme: a plant active oxygen rapid detection kit, comprising Staining solution, H2O2 staining solution and decolorizing solution, the The staining solution is 0.5 mol / ml nitro blue tetrazolium chloride, the H2O2 staining solution is 0.1 mol / ml diaminobenzidine, and the decolorizing solution is a mixed solution prepared by mixing anhydrous ethanol and glacial acetic acid in a mass ratio of 3:1.

[0006] A rapid detection kit for plant reactive oxygen species and H2O2 application, comprising the following steps:

[0007] Step 1: dyeing treatment, cutting off fresh plant samples and immersing them in dyeing liquid for dyeing treatment;

[0008] Step 2: Decolorization: add decolorizing solution to a 2 ml centrifuge tube containing the stained sample, and then place it in a boiling water bath and heat it until the sample becomes colorless and then take it out;

[0009] Step 3: Observe and take pictures. Place the decolorized sample on a glass slide and add glycerol to fix it. Cover it with a coverslip and move it under a microscope for observation and photography.

[0010] A further improvement is that: in step 1, When staining with the staining solution, stain at 25°C in a dark environment for 2 hours. When staining with H2O2 staining solution, stain at 25°C in a dark environment for 8 hours.

[0011] Further improvements are: the water bath heating time in step 2 is 30 minutes; and glycerol with a mass percentage concentration of 20% is used in step 3.

[0012] The beneficial effects of the present invention are as follows: the kit of the present invention can quickly detect plant active oxygen, the process is fast and time-saving, the operation is simple, the whole process only needs to use a water bath pot and a microscope for detection, the cost is low, and the active oxygen in plants can be seen. And the real-time distribution of H2O2 in the roots and leaves of plants. At the same time, the kit can be widely used in herbaceous plants and woody plants, with a wide range of applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 The edge of the grape leaf in Example 1 of the present invention Staining result diagram.

[0014] Figure 2 The central vein of the grape leaf of Example 1 of the present invention Staining result diagram.

[0015] Figure 3 The grape root of Example 2 of the present invention Staining result diagram.

[0016] Figure 4 The rape root of Example 2 of the present invention Staining result diagram.

[0017] Figure 5 This is a diagram showing the H2O2 staining results of grape leaves in Example 3 of the present invention. DETAILED DESCRIPTION

[0018] In order to deepen the understanding of the present invention, the present invention will be further described in detail below in conjunction with examples. The examples are only used to explain the present invention and do not constitute a limitation on the protection scope of the present invention.

[0019] This embodiment provides a plant active oxygen rapid detection kit, including Staining solution, H2O2 staining solution and decolorizing solution, The staining solution is 0.5 mol / ml nitro blue tetrazolium chloride, the H2O2 staining solution is 0.1 mol / ml diaminobenzidine, and the decolorizing solution is a mixture of anhydrous ethanol and glacial acetic acid in a mass ratio of 3:1.

[0020] Example 1

[0021] according to Figure 1 and Figure 2 As shown, this embodiment provides a plant active oxygen rapid detection kit for grape leaves The dyeing application includes the following steps:

[0022] Step 1: Dyeing treatment: Cut fresh grape leaf samples and immerse them in The staining was carried out in the staining solution at 25°C in a dark environment for 2 h;

[0023] Step 2: Decolorization: add decolorizing solution to a 2 ml centrifuge tube containing the stained sample, and then place it in a boiling water bath and heat it for 30 minutes until the sample becomes colorless and then take it out;

[0024] Step 3: Observe and take photos. Place the decolorized sample on a glass slide and add 20% glycerol to fix it. Cover it with a coverslip and move it under a microscope for observation and photography.

[0025] Instructions attached Figure 1 And the instruction manual Figure 2 Around the edges and central veins of grape leaves Staining result diagram.

[0026] Example 2

[0027] according to Figure 3 and Figure 4 As shown, this embodiment provides a plant active oxygen rapid detection kit for the root of grape and rapeseed The dyeing application includes the following steps:

[0028] Step 1: Dyeing treatment: Cut fresh samples of grape and rape roots and immerse them in The staining was carried out in the staining solution at 25°C in a dark environment for 2 h;

[0029] Step 2: Observe and take photos. Place the stained sample on a glass slide and add 20% glycerol to fix it. Cover it with a coverslip and move it under a microscope for observation and photography.

[0030] Instructions attached Figure 3 For grape roots Staining result diagram, instruction manual attached Figure 4 O2 for rapeseed roots - Staining result diagram.

[0031] Example 3

[0032] according to Figure 1 and Figure 2 As shown, this embodiment provides an application of a plant active oxygen rapid detection kit in H2O2 staining of grape leaves, comprising the following steps:

[0033] Step 1: dyeing treatment: cut fresh grape leaf samples, immerse them in H2O2 dyeing solution, and dye them for 8 hours in a dark environment at 25°C;

[0034] Step 2: Decolorization: add decolorizing solution to a 2 ml centrifuge tube containing the stained sample, and then place it in a boiling water bath and heat it for 30 minutes until the sample becomes colorless and then take it out;

[0035] Step 3: Observe and take photos. Place the decolorized sample on a glass slide and add 20% glycerol to fix it. Cover it with a coverslip and move it under a microscope for observation and photography.

[0036] Instructions attached Figure 5 This is the result of H2O2 staining of grape leaves.

[0037] The above shows and describes the basic principles, main features and advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A rapid detection kit for plant active oxygen, characterized in that: include Staining solution, H2O2 staining solution and decolorizing solution, the The staining solution is 0.5 mol / ml nitro blue tetrazolium chloride, the H2O2 staining solution is 0.1 mol / ml diaminobenzidine, and the decolorizing solution is a mixed solution prepared by mixing anhydrous ethanol and glacial acetic acid in a mass ratio of 3:

1.

2. A rapid detection kit for plant reactive oxygen species and H2O2, characterized in that The following steps are involved: Step 1: dyeing treatment, cutting off fresh plant samples and immersing them in dyeing liquid for dyeing treatment; Step 2: Decolorization: add decolorizing solution to a 2 ml centrifuge tube containing the stained sample, and then place it in a boiling water bath and heat it until the sample becomes colorless and then take it out; Step 3: Observe and take pictures. Place the decolorized sample on a glass slide and add glycerol to fix it. Cover it with a coverslip and move it under a microscope for observation and photography.

3. A plant active oxygen rapid detection kit according to claim 2 is used to detect plant active oxygen and H2O2, characterized in that: In step 1, use When staining with the staining solution, stain at 25°C in a dark environment for 2 hours. When staining with H2O2 staining solution, stain at 25°C in a dark environment for 8 hours.

4. A plant active oxygen rapid detection kit according to claim 2 for detecting plant and H2O2, characterized in that: The water bath heating time in step 2 is 30 minutes; and the glycerol used in step 3 is 20% by mass percent.

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