Method for rapidly detecting wheat tissue callose

A detection method, callose technology, is applied in the field of rapid detection of wheat tissue callose, which can solve the problems of expensive equipment, affecting the production effect, and difficulty in mastering the technology, and achieves easy mastery, strong operability, and relative content difference obvious effect

Inactive Publication Date: 2013-06-12
CROP RES INST SHANDONG ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the cryosection technology is relatively mature in the slice production of animal materials (because the cells do not have cell walls), it is difficult to master the technology in plant tissues, and the equipment is expensive and the cost of consumables is high. What is more serious is that the material is difficult to fix On the base, when cutting materials with a thick silicon layer such as wheat and rice, it is easy to cause the knife to vibrate, which affects the production effect

Method used

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  • Method for rapidly detecting wheat tissue callose
  • Method for rapidly detecting wheat tissue callose
  • Method for rapidly detecting wheat tissue callose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A rapid detection method for callose in wheat tissue, the application in the leaf tissue of wheat variety Jimai 22, the specific operation is as follows:

[0033] Prepare the solution:

[0034] Preparation of dyeing loading buffer: prepare 100mmol / L Tris-HCl buffer (pH=8.8); then dissolve 0.01g of water-soluble aniline blue in 100ml of the above buffer to obtain the dyeing loading buffer.

[0035] Material processing: Wheat leaf tissue in the mid-filling stage was taken, then brought back to the laboratory, and blotted dry with absorbent paper.

[0036] Freehand slicing: use a sharp blade (surgical blade) to cut the wheat leaf tissue into slices with a thickness of 20-40 μm, and place them on a glass slide;

[0037] Material staining: Use a pipette gun or a pipette to draw 100-150 μL of sample staining buffer and drop it on the slide placed on the slide, then place the slide into a vacuum container, seal it and pump it with a vacuum pump 5-8 minutes to remove the air ...

Embodiment 2

[0042] A rapid detection method for callose in wheat tissue, the application in the stem tissue of wheat variety Jimai 22, the specific operation is as follows:

[0043] Prepare the solution:

[0044] Preparation of dyeing loading buffer: prepare 150mmol / L K2HPO4 buffer (pH=9.5); then dissolve 0.01g of water-soluble aniline blue in 100ml of the above buffer to obtain the dyeing loading buffer;

[0045] Material processing: Wheat stem tissues in the mid-filling stage were taken, then brought back to the laboratory, and blotted dry with absorbent paper.

[0046]Freehand slicing: use a sharp blade (surgical blade) to cut the wheat leaf tissue into slices with a thickness of 20-40 μm, and place them on a glass slide;

[0047] Material staining: Use a pipette gun or a pipette to draw 100-150 μL of sample staining buffer and drop it on the slide placed on the slide, then place the slide into a vacuum container, seal it and pump it with a vacuum pump 5-8 minutes to remove the air b...

Embodiment 3

[0052] A rapid detection method for callose in wheat tissue, under the conditions of fixative fixation for 24 hours, conservation tillage (the corn stalks of the last crop crop were directly crushed and returned to the field after harvest, wheat was sown with a no-till machine), and different nitrogen application rates The application in the stem tissue of Jimai 22 at the flowering stage, the specific operation is as follows:

[0053] Prepare the solution:

[0054] Preparation of fixative: Mix ethanol and chloroform at 3:1 (v / v) as a mixed solvent, and use the above mixed solvent as a solvent to prepare a 0.15% trichloroacetic acid solution;

[0055] Preparation of dye loading buffer: prepare 67mmol / L of K 2 HPO 4 Buffer solution (pH=9.0); then dissolve 0.01g of water-soluble aniline blue in 100ml of the above buffer solution to be the dye loading buffer solution;

[0056] Material processing: Wheat stem tissues in the mid-filling stage were taken, then brought back to the ...

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Abstract

The invention discloses a method for dyeing and observing wheat tissue callose and belongs to the technical field of chemical plant tissue positioning. The method mainly comprises the following steps: (1) preparing a solution; (2) treating the material; (3) slicing; (4) dyeing materials; (5) producing sections; and (6) exciting through a laser excitation device, and observing and shooting through a fluorescence microscope. By adoption of a bare-handed slicing method, the wheat tissue slices which are manufactured in a bare-handed mode are dyed under the alkaline conditions by using the callose specifity fluorescent dye-water-soluble aniline blue, the sample slice manufacturing, dyeing and observing period is obviously shortened, and the callose is accurately positioned. The method is high in operability, the manufactured slices are conveniently observed, and the acquired image is clear.

Description

technical field [0001] The invention relates to the technical field of microscopic preparation of plant tissue and chemical localization of tissue components, in particular to a rapid detection method for callose in wheat tissue. Background technique [0002] Callose is a kind of glucan bound by β-1,3-bonds, which plays an important role in regulating plant life activities such as sieve tube metabolism and gametophyte development. Its synthesis and decomposition are directly related to the normal growth and metabolism of plants. process. In addition, when plants are subjected to adverse environmental stress or strong stimuli, such as pathogenic bacteria infection, nutritional stress, mechanical damage, high temperature and other adversities, callose metabolism will immediately feedback and regulate the physiological activities of plant cells (Wang, McCallum, Fetch et al. 2012; Chapman et al., 2009; Saulnier et al., 2012). Under adversity, callose will be quickly synthesize...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N1/30
Inventor 孔令安
Owner CROP RES INST SHANDONG ACAD OF AGRI SCI
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