Paraffin sectioning method for peanut seeds

A technology of paraffin sectioning and seed kernels, which is applied in the preparation of test samples, etc., can solve the problems of unsatisfactory paraffin sectioning effects and no reports, and achieve the effect of easy popularization and application and simple operation

Active Publication Date: 2013-02-06
SHANDONG PEANUT RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the high oil and protein content of peanut kernels, the effect of its paraffin section has been unsatisfactory
[0005] Therefore, in order to facilitate the observation of peanut cell structure and dynamic changes, deeply analyze the cytological mechanism of Aspergillus flavus infection in peanuts, clarify the cytological differences between resistant and susceptible peanut germplasms, and screen peanut germplasms resistant to Aspergillus flavus for production practice, Need a kind of paraffin section method that can obtain the peanut kernel of ideal section badly, but there is no report yet about this kind of section method at present

Method used

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  • Paraffin sectioning method for peanut seeds
  • Paraffin sectioning method for peanut seeds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1. Material: Take the peanut kernels of resistant and susceptible peanut varieties J11 and Jinhua 1012 8 days (days) after Aspergillus flavus infection, separate two cotyledons, and then cross-cut them.

[0052] 2. Fixation: Fix commonly used glutaraldehyde with electron microscope samples, the volume concentration of glutaraldehyde is 5%, double distilled water as solvent, put peanut kernel slices into fixative, and replace fixative 3 times during fixation .

[0053] 3. Dehydration: The samples are dehydrated by ethanol. The specific dehydration procedures are: 80% ethanol for 50 minutes, 95% ethanol for 45 minutes, and 100% ethanol for 45 minutes. Each step of the dehydration process is two times. Before the sample is dehydrated, the sample is wrapped with section paraffin at 54~56℃. Buried positioning is convenient for future operations.

[0054] 4. Transparency: Use 1:1 (volume ratio) xylene:ethanol transition method. The specific transparent process is as follows: 1:1 xy...

Embodiment 2

[0061] 1. Material: Take the peanut kernels of resistant and susceptible peanut varieties J11 and Jinhua 1012 2d, 4d, 6d, 8d, 10d after Aspergillus flavus infection, separate two cotyledons, and then cross-cut them.

[0062] 2. Fixation: Fix the commonly used glutaraldehyde with the electron microscope sample, the volume concentration of glutaraldehyde is 5%, double distilled water as the solvent, put the peanut kernel slices into the fixative, and replace the fixative 3 times during the fixation process .

[0063] 3. Dehydration: The samples are dehydrated by ethanol. The specific dehydration procedures are: 80% ethanol for 50 minutes, 95% ethanol for 45 minutes, and 100% ethanol for 45 minutes. Each step of the dehydration process is performed twice. Before the sample is dehydrated, the sample is wrapped in paraffin at 54-56°C. Buried positioning is convenient for future operations.

[0064] 4. Transparency: 1:1 (volume ratio) xylene:ethanol transition method is adopted. The speci...

Embodiment 3

[0071] 1. Material: Take the peanut kernels of resistant and susceptible peanut varieties J11 and Jinhua 1012 2d, 4d, 6d, 8d, 10d after Aspergillus flavus infection, separate two cotyledons, and then cross-cut them.

[0072] 2. Fixation: Fix commonly used glutaraldehyde with electron microscope samples, the volume concentration of glutaraldehyde is 10%, double distilled water as solvent, put peanut kernel slices into fixative, and replace fixative 4 times during fixation .

[0073] 3. Dehydration: The samples are dehydrated by ethanol. The specific dehydration procedures are: 80% ethanol for 50 minutes, 95% ethanol for 45 minutes, and 100% ethanol for 45 minutes. Each step of the dehydration process is performed twice. Before the sample is dehydrated, the sample is wrapped in paraffin at 54-56°C. Buried positioning is convenient for future operations.

[0074] 4. Transparency: Adopt 1:1 (volume ratio) xylene:ethanol transition method. The specific transparent process is as follows: ...

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Abstract

The invention provides a paraffin sectioning method for peanut seeds. The method includes the steps of 1) material selection; 2) fixation by glutaraldehyde 5%-10% in concentration; 3) dehydration: dehydrating by ethanol 80% in concentration for 50min, ethanol 95% in concentration for 45min and ethanol 100% in concentration for 45min and repeating each gradient for one time; 4) transparency: treating by ethanol 100% in concentration for 20-30min, and treating by dimethylbenzene for 15-25min twice, wherein the volume ratio of the dimethylbenzene to the ethanol is 1:1; 5) paraffin permeation by a direct paraffin permeation method, embedding and sectioning; and 6) deparaffinage, rehydration, hematoxylin-eosin redyeing, dehydration, transparency and sealing. The method is capable of well fixing cellular tissue of the peanut seeds, complete and continuous in sectioning, free of cavities, simple to operate, applicable to all peanut varieties and convenient to popularize and apply, and section thickness can reach 7 micrometres.

Description

【Technical Field】 [0001] The present invention relates to the technical field of paraffin sectioning, in particular to a paraffin sectioning method of peanut kernels. 【Background technique】 [0002] Peanut kernels are the main target product of peanut harvest, which determines the formation of peanut yield and quality, but peanut kernels are susceptible to infection by Aspergillus flavus and other pathogens during the growth process. Studies have shown that the characteristics of low permeability, toughness and damage resistance of peanut seed coats make peanuts passively resistant to diseases and insect pests. Therefore, the integrity and strength of peanut seed coats have an important barrier to resistance to Aspergillus flavus. effect. [0003] Peanut seed coat structure is an important indicator to evaluate the resistance and susceptibility of peanut germplasm to Aspergillus flavus infection. Ordinary microscopic observation shows that the seed coat structure of disease-resist...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 单世华闫彩霞李春娟张廷婷许婷婷
Owner SHANDONG PEANUT RES INST
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