Method for obtaining vascular cambium of woody plant and application thereof

A woody plant and cambium technology, applied in the biological field, can solve the problem of little understanding of molecular regulation mechanism, and achieve the effect of high cell homogeneity and complete cell structure

Active Publication Date: 2020-03-27
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, how to obtain high-quality vascular cambium in woody plants, especially fast-growing tree species, is still a technical problem, and little is known about the molecular regulation mechanism of vascular cambium formation and cell division in woody plants

Method used

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  • Method for obtaining vascular cambium of woody plant and application thereof
  • Method for obtaining vascular cambium of woody plant and application thereof
  • Method for obtaining vascular cambium of woody plant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1. Combination of paraffin section and laser microdissection technology to accurately obtain the vascular cambium of the second stem node of P.

[0044] 1. Preparation of Paraffin Sections

[0045] 1. Material preparation: Transplant the aseptic test-tube seedlings of P. chinensis after one month of growth into the soil, and continue to cultivate them in the greenhouse for four months before using them as experimental materials. The growth conditions for greenhouse culture are: light intensity 1000-1500lx, day and night time 12h / 12h, culture temperature (25±2)°C.

[0046] 2. Take the well-grown yellow beam wood and transplant the seedlings for four months. figure 1 As shown, the leaves were cut off with a single-sided blade wiped with RNaseZap, leaving the second stem node, and each stem node was cut into small sections of 2-4 mm, and immediately put into the fixative. During the sampling process, try to reduce the RNAase that exists during the experiment. The...

Embodiment 2

[0067] Example 2. The combination of paraffin section and laser microdissection technology to accurately obtain the vascular cambium of the fourth stem node of Huangliang wood

[0068] 1. Preparation of Paraffin Sections

[0069] 1. Material preparation: Transplant the aseptic test-tube seedlings of P. chinensis after one month of growth into the soil, and continue to cultivate them in the greenhouse for four months before using them as experimental materials. The growth conditions for greenhouse culture are: light intensity 1000-1500lx, day and night time 12h / 12h, culture temperature (25±2)°C.

[0070] 2. Take the well-grown yellow beam wood and transplant the seedlings for four months. figure 1As shown, the leaves were cut off with a single-sided blade wiped with RNaseZap, leaving the fourth stem node, and each stem node was cut into small sections of 2-4 mm, and immediately put into the fixative. During the sampling process, try to reduce the RNAase that exists during the...

Embodiment 3

[0089] Example 3. The combination of frozen section and laser microdissection technology to accurately obtain the study of the vascular cambium of yellow beam wood

[0090] (1) Frozen section quick freezing method

[0091] The four-month-old seedlings are transplanted from well-grown yellow beam wood, and the parts of the materials are as follows: figure 1 As shown, cut off the leaves with a single-sided blade wiped with RNaseZap, and leave the second stem node. Each stem node is cut into small sections of 2-4mm, and the small stem sections are put into a 1.5mL centrifuge tube and put into liquid nitrogen. Quickly freeze for 60 seconds, use tweezers to take out the centrifuge tube and put it into the cryostat box to warm up for about 20 minutes, half-wrap the material with OCT embedding medium and fix it on the sample top, cutting the thickness of 8-14 μm. Use tweezers to paste the excised tissue on the glass slide, and use a pipette gun to drop 100% ethanol on the glass slid...

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Abstract

The invention provides a method for obtaining a vascular cambium of a woody plant and application thereof. According to the method, the vascular cambium in a woody plant tissue paraffin section containing the vascular cambium is obtained through a laser microdissection method. According to the method, the pure cambium tissue of the woody plant neolamarckia cadamba is successfully separated throughorganic combination of paraffin section and laser microdissection technologies. According to the method, the paraffin section process is optimized, LMD parameters are debugged, a laser microdissection technical system of the vascular tissue cells of the seedlings of the neolamarckia cadamba is established, and a high-quality vascular cambium is obtained. The application of the method disclosed bythe invention in similar tissues of woody plants is not reported, the tissue and organ cells obtained by the method are relatively high in homogeneity and complete in cell structure, and a foundationis laid for further researching molecular biology and genomics research of cambium.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for obtaining vascular cambium of woody plants and its application. Background technique [0002] The radial growth of wood is the result of the inward differentiation of the vascular cambium into secondary xylem and outward differentiation into secondary phloem. The vascular cambium is crucial for the growth of woody plants. It is the difference between forest trees and other annual or The major tissue of herbaceous perennials, cambium activity, leads to thickening of stems with a concomitant increase in biomass. In addition, the vascular cambium can also form callus in the injured part of the plant to protect itself, and dormant in the harsh growth environment to avoid cell damage. Therefore, the study of the gene expression characteristics of vascular cambium and the molecular genetic mechanism of the division and differentiation of cambium cells will lay an important fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/06G01N1/36
CPCG01N1/06G01N1/36
Inventor 张建霞王雪彭昌操赵小兰龙健梅
Owner SOUTH CHINA AGRI UNIV
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