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A method for in situ detection of hemicellulose distribution in plant cell walls

A technology of hemicellulose and plant cells, which is applied in the field of in situ detection of hemicellulose distribution in plant cell walls, can solve the problems of long sampling period, incomplete removal of hemicellulose, hemicellulose removal, etc., to achieve high efficiency The effect of selective separation

Active Publication Date: 2016-05-11
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of this method are: the research object is a single fiber, the size is small, the chemical composition content is low, and there are many technical difficulties
The disadvantage of this method is that part of the hemicellulose has been removed during the preparation of the hemicellulose; the hemicellulose cannot be completely removed from the hemicellulose
The disadvantages of this method are: the sampling period is long, the sampling location should be representative and the size is small, the chemical composition content is low, and there are great difficulties in separation technology
And it cannot reflect the in situ distribution of different kinds of hemicelluloses in each microdomain of the cell wall

Method used

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  • A method for in situ detection of hemicellulose distribution in plant cell walls
  • A method for in situ detection of hemicellulose distribution in plant cell walls
  • A method for in situ detection of hemicellulose distribution in plant cell walls

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Embodiment 1

[0040] The embodiment of the present invention provides a method for in situ detection of the distribution of hemicellulose in each micro-region of the plant cell wall, the method comprising:

[0041] Step 1: Cut poplar tissue into small strips of 0.5mm (length) x 0.5mm (width) x 5mm (height);

[0042] The smaller the size of the poplar wood strips, the time required for subsequent fixation, dehydration and infiltration will be shortened accordingly; but when the size of the poplar wood strips is too small, it is difficult to control the poplar wood strips in the transparent capsule during subsequent embedding. When the size of the poplar wood strip is 0.5mm (length) x0.5mm (width) x5mm (high), it can ensure that the poplar wood strip can be fixed, dehydrated and infiltrated quickly, and can keep the poplar wood strip. The position of the small wooden strips in the transparent capsule is beneficial to improve the quality of ultrathin sections.

[0043] Step 2: Immerse the cut...

Embodiment 2

[0068] The embodiment of the present invention provides a method for in situ detection of the distribution of hemicellulose in each micro-region of the plant cell wall, the method comprising:

[0069] Step 1: Cut poplar tissue into small strips of 0.5mm (length) x 0.5mm (width) x 5mm (height);

[0070] Step 2: Immerse the cut poplar wood strips in a fixative solution mixed with 4% paraformaldehyde + 0.1% glutaraldehyde for fixation. 4 hours under normal pressure;

[0071] Step 3: The fixed poplar wood strips were immersed in 50 mM sodium cacodylate buffer solution for washing, and kept at room temperature and normal pressure for 10 min. The number of washes is 3 times;

[0072] Step 4:

[0073] The washed plant tissue pieces were immersed in 30v / v% ethanol solution once, 50v / v% ethanol solution once, 70v / v% ethanol solution once, 80v / v% ethanol solution once, 90v / v% ethanol solution Water solution 1 time, 95v / v% ethanol water solution 3 times, 100% ethanol 3 times for dehy...

Embodiment 3

[0096] The embodiment of the present invention provides a method for in situ detection of the distribution of hemicellulose in each micro-region of the plant cell wall, the method comprising:

[0097] Step 1: Cut the awn tissue into small wooden strips of 0.5mm (length) x 0.5mm (width) x 5mm (height);

[0098] Step 2: Immerse the cut thin Miscanth strips in a fixative solution mixed with 4% paraformaldehyde + 0.1% glutaraldehyde for fixation. At room temperature, first keep it under the condition of vacuum degree of -0.098MPa for 40min, then Keep at normal pressure for 4 hours;

[0099] Step 3: The fixed strips of Miscanthus acanthus were immersed in 50 mM sodium cacodylate buffer solution for washing, and kept at room temperature and normal pressure for 10 min. The number of washings is 3 times;

[0100] Step 4:

[0101] Immerse the washed Miscanthus strips in 30v / v% ethanol solution once, 50v / v% ethanol solution once, 70v / v% ethanol solution once, 80v / v% ethanol solution ...

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Abstract

The invention discloses a method for detecting distribution of hemicellulose on plant cell walls in situ. The method comprises the following steps: firstly, performing fixing, washing, dehydrating, permeating and embedding polymerization treatment on plant tissue blocks; subsequently, sectioning the embedded plant tissue blocks to obtain ultrathin sections and collecting the ultrathin sections on a nickel net; subsequently, performing washing, sealing and first antibody incubation treatment on a carrying side of the nickel net; washing the carrying side of the nickel net after first antibody incubation, and performing second antibody incubation; and finally, dyeing the carrying side of the nickel net and observing the carrying side of the nickel net by an electron microscope. According to the method, hemicellulose of different varieties can be effectively identified by a first antibody which can be specifically combined with the hemicellulose of different varieties, so that the in-situ distribution of the hemicellulose of different varieties in microcells of the plant cell walls can be accurately and effectively observed.

Description

technical field [0001] The invention relates to the technical field of plant ultrastructure detection, in particular to a method for in situ detection of the distribution of hemicellulose in plant cell walls. Background technique [0002] Energy shortage, environmental pollution, food and medicine safety are the great challenges facing the world today, and the development of clean and safe biomass resources is an effective way to deal with these challenges. As the second most abundant renewable biomass resource in nature, hemicellulose can be widely used in chemical, food, papermaking, pharmaceutical and coating industries, and has broad development prospects and huge social, ecological and economic benefits. At present, the high-value comprehensive utilization of hemicellulose first needs to be efficiently and selectively separated from biomass raw materials. However, due to the close combination or chemical bonds between hemicellulose and other components in biomass raw m...

Claims

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Application Information

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IPC IPC(8): G01N23/04
Inventor 许凤马静周霞吉喆张逊
Owner BEIJING FORESTRY UNIVERSITY