Wheat tissue active oxygen fluorescence labeling method

A fluorescent labeling, reactive oxygen species technology, applied in the field of plant stress and physiological response, can solve the problems of lack of scientific detection method for the comprehensive production of reactive oxygen species, large differences in production, difficult plant stress resistance or aging process, etc. Achieve precise resistance or aging process, easy operation, improve the effect of fixing and loading

Inactive Publication Date: 2012-07-18
CROP RES INST SHANDONG ACAD OF AGRI SCI
View PDF9 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many kinds of reactive oxygen species in plant tissues, and their production varies greatly. Therefore, it is difficult to explain the stress resistance or aging process of plants with the chang

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Wheat tissue active oxygen fluorescence labeling method
  • Wheat tissue active oxygen fluorescence labeling method
  • Wheat tissue active oxygen fluorescence labeling method

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0028] Example 1

[0029] A fluorescent labeling method for plant tissue reactive oxygen species, which is used in wheat (Jimai 22) stem tissue under nutrient stress (nitrogen deficiency). The specific operations are as follows:

[0030] a. Prepare wheat tissue sample labeling and loading buffer, draw 5 ml of sample labeling and loading buffer into a container with a volume of 10 ml;

[0031] Prepare 10 mM Tris-HCl buffer (pH 7.2); prepare DCFH-DA stock solution with DMSO at a concentration of 5 mM; prepare fluorescent label loading solution with buffer, containing DCFH-DA at a final concentration of 50 μM and a final concentration of 50 mM KCl and glutaraldehyde at a final concentration of 2.5% (volume percentage).

[0032] b. Cut wheat stalks into tissue pieces with a length of 0.5 cm, and cut them evenly along the longitudinal axis, and divide them into two;

[0033] c. Rinse the tissue block made in step b with the sample labeling loading buffer twice, and then place it in the vess...

Example Embodiment

[0039] Example 2

[0040] A method for fluorescent labeling of plant tissues with reactive oxygen species, applied to leaf tissues of wheat (Jimai 22) during grain-filling and senescence, the specific operations are as follows:

[0041] a. Prepare wheat tissue sample labeling and loading buffer, draw 5 ml of sample labeling and loading buffer into a container with a volume of 10 ml;

[0042] The preparation method of the above labeling loading buffer is: preparing 10 mM PBS buffer (pH 7.4); preparing DCFH-DA stock solution with ethanol at a concentration of 3 mM; preparing fluorescent labeling loading buffer with buffer, containing a final concentration of 30 μM DCFH-DA, a final concentration of 50 mM KCl and a final concentration of 2.5% (volume percentage) glutaraldehyde;

[0043] b. Cut the wheat leaves into tissue pieces with a length of 0.5 cm;

[0044] c. Rinse the tissue block prepared in step (2) with the sample labeling load buffer twice, then place it in the vessel prepared i...

Example Embodiment

[0048] Example 3

[0049] A fluorescent labeling method for plant tissue reactive oxygen species, the application of wheat (Jimai 22) leaf tissue under drought stress, the specific operations are as follows:

[0050] a. Prepare wheat tissue sample labeling and loading buffer, draw 5 ml of sample labeling and loading buffer into a container with a volume of 10 ml;

[0051] Prepare 10 mM PBS buffer (pH 7.2); prepare DCFH-DA stock solution with methanol at a concentration of 3 mM; prepare fluorescent label loading solution with buffer, containing DCFH-DA at a final concentration of 30 μM and a final concentration of 50 mM KCl and glutaraldehyde with a final concentration of 2.2-2.8% (volume percentage).

[0052] b. Cut the wheat leaves into tissue pieces with a length of 0.5 cm;

[0053] c. Rinse the tissue block prepared in step (2) with the sample labeling load buffer twice, then place it in the vessel prepared in step (1), seal and pump air, so that all the plant tissue blocks sink to ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Lengthaaaaaaaaaa
Login to view more

Abstract

The invention belongs to the technical field of plant stress and physiologic response, and particularly relates to the labeling method for active oxygen generation, generation positions and relative content under biotic stress and abiotic stress during the growth and development process of wheat. The labeling method comprises the following steps of: preparing a label load buffer solution, preparing a material tissue block, rinsing the tissue block with the load buffer solution, placing the tissue block in a dark place at 25 DEG C, exciting the tissue block by using a laser excitation device, observing the tissue block and taking a photo. Compared with the traditional labeling method of DAB (Dimethylaminoazobenzene), NBT (Nitroblue Tetrazolium) and other active oxygen species, the labeling method saves time and avoids the distortion of active oxygen content and positions of a sample during long-time labeling load period. When the active oxygen is labeled by DCFH-DA (Dichlorofluorescein Diacetate), the material is fixed, and the accuracy of active oxygen labeling is ensured. The used device has simple requirements and is easy to operate.

Description

technical field [0001] The invention belongs to the technical field of plant adversity stress and physiological response, and relates to a fluorescent labeling method for the location and relative content detection of active oxygen in plant tissues, and in particular to a method for detecting active oxygen under biotic and abiotic stress during the growth and development of wheat Marking method of production, production site and relative content. Background technique [0002] During the growth and development of plants, they are constantly subjected to various adversities and the inevitable aging process, which leads to metabolic disorders of reactive oxygen species in plant cells, that is, reactive oxygen species in plants such as superoxide anions (OH· - ), hydrogen peroxide (H 2 o 2 ), hydroxyl radical (-OH), singlet oxygen ( 1 o 2） and other bursts; and active oxygen scavengers, such as superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), carotenoids, vit...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N1/28
Inventor 孔令安王法宏冯波李升东张宾司纪升
Owner CROP RES INST SHANDONG ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap