Method for testing lignification degrees of plant cell walls

A measurement method and plant cell technology, which is applied in the field of confocal Raman microscopy to determine the degree of lignification of plant cell walls, can solve the problems of no research reports on the degree of lignification of plant cell walls under confocal microscopy, etc., to reduce the interference of fluorescence, reduce the Fluorescence interference, the effect of ensuring accuracy

Inactive Publication Date: 2012-05-02
BEIJING FORESTRY UNIVERSITY
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There is no domestic patent on the study of differences in the degree of lignification of plant cell walls using Raman microscopy
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  • Method for testing lignification degrees of plant cell walls
  • Method for testing lignification degrees of plant cell walls
  • Method for testing lignification degrees of plant cell walls

Examples

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Embodiment 1

[0047] Study on the lignification process of different morphological regions of the fiber cell wall of Cornus alba

[0048] 1. Preparation of softening material

[0049] Cut Hongduanmu wood into cuboids with a size of 1 cm (along the grain direction) × 0.5 cm (chord direction) × 1 cm (radial direction, thickness direction) with a single-sided blade, and then soak them in boiling water (100°C) for 45 minutes to obtain softening material;

[0050] 2. Preparation of sample slices

[0051] Cut the softened material into a cross-section with a thickness of 20±2 μm along the radial direction of the softened material with a slide-away microtome to obtain a thin slice of the sample;

[0052] 3. Eliminate fluorescence interference

[0053] Soak the prepared sample sheet in sodium borohydride solution for 5 minutes, the mass percent concentration of sodium borohydride solution is 1%, sodium borohydride reacts with the carbonyl group in the sample sheet plant cells, the carbonyl group...

Embodiment 2

[0058] In addition to using Hongduanmu as raw material, to study the degree of lignification of the secondary wall of the vessel, the secondary wall of the ray parenchyma cell, and the secondary wall of the fiber cell, the soaking temperature of the preparation step of the softening material is 95, and the soaking time is 40 minutes; the thickness of the sample sheet It is 25±2 μm; without the step of eliminating fluorescence interference, that is, without soaking the sample sheet with sodium borohydride solution, the rest is the same as in Example 1.

[0059] At 250~3250cm -1 Collect spectrum within the range, for 1580~1640cm -1 Ranges are integrated where 1606cm -1 The characteristic peak of the stretching vibration of lignin benzene ring C=C. 1655cm -1 The stretching vibration of coniferyl aldehyde C=O and the stretching vibration of coniferyl alcohol C=C are characteristic peaks.

[0060] The Raman images of the secondary wall of red endwood vessels, the secondary wall...

Embodiment 3

[0062] In addition to using Pinus yunnanensis as raw material to study the degree of lignification of different morphological regions of its fiber cells, the soaking time for the preparation of softened wood was 30 minutes, and the concentration of sodium borohydride solution in the step of eliminating fluorescence interference was 1.5%. Soaking time is except 7min, and all the other are identical with embodiment 1.

[0063] Raman microscope image of the stressed wood fiber cell wall of Pine yunnanensis image 3 Shown: from image 3 It can be seen that the lignification degree of the outer layer 6 of the secondary wall of the fibroblast near the cell corner is the highest, and the calculated value of the Gauss-Lorentz mixed function of the Raman signal intensity reaches 3000; the second is the cell corner 1, and its The Gauss-Lorentz mixed function calculated value of the Raman signal intensity is 2500; the calculated value of the Gauss-Lorentz mixed function of the Raman sig...

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Abstract

The invention discloses a method for testing lignification degrees of plant cell walls based on a confocal Raman microscopic technology. The method comprises the following steps of: testing Raman spectrums of the plant cell walls under the in-situ state by using a confocal Raman microscope to obtain Raman spectrum imaging graphs of lignin in the cell walls; and determining the content of the lignin in the cell walls according to Raman signal intensities, and determining the lignification degrees of different secondary cell walls and different form areas of fiber cell walls. The testing method is based on a pure spectrum method, samples are quickly prepared, chemical pollutants are not generated, and the information of the lignification degrees of the plant cell walls can be acquired under the in-situ state.

Description

technical field [0001] The invention relates to the application of Raman spectroscopy combined with confocal microscope technology in the study of lignification degree of plant cell walls, in particular to a method for measuring the lignification degree of plant cell walls by using a confocal Raman microscope with an excitation wavelength of 785nm in an in situ state. Background technique [0002] Lignocellulosic raw materials are considered as fuel and chemical resources for sustainable recycling. Its cell wall is a hard layered structure similar to reinforced concrete composed of cellulose, hemicellulose and lignin. These three polymer macromolecules account for More than 90% of the total cell wall biomass. Among them, lignin is a complex and irregular polyphenolic polymer existing in vascular plants, which enhances the structural integrity of cell walls and also inhibits microbial corrosion of cell walls. The complexity of its structure and function has always been the b...

Claims

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Application Information

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IPC IPC(8): G01N21/65G01N1/28
Inventor 马建锋张智衡毛健贞许凤孙润仓
Owner BEIJING FORESTRY UNIVERSITY
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