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A Rapid Histochemical Method for Visualizing Laticifers in Rubber Tree Bark

A rubber tree and bark technology, which is applied in the preparation of test samples, material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of difficult repeatability of slice quality, potential safety hazards, easy to make mistakes in labels, etc., to achieve repeatability and Universal batch screening, convenient material collection, time-saving effect

Active Publication Date: 2019-10-11
RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method of displaying milk ducts has two major disadvantages: 1) the steps are numerous, time-consuming, easy to mislabel, and the quality of slices is not easily repeatable
In addition, multiple steps are operated at high temperature, posing safety hazards

Method used

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  • A Rapid Histochemical Method for Visualizing Laticifers in Rubber Tree Bark
  • A Rapid Histochemical Method for Visualizing Laticifers in Rubber Tree Bark
  • A Rapid Histochemical Method for Visualizing Laticifers in Rubber Tree Bark

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Embodiment 1, the primary milk duct histochemical display of bark under the natural state of annual saplings

[0031] In this embodiment, the bark of the young bark of the rubber tree variety CATAS7-33-97 in a natural state is used for histochemical display of the primary laticiferous ducts of the bark.

[0032]1. Experimental conditions: the bark of the new saplings of the rubber tree variety CATAS7-33-97 in the stable period was trimmed and cut into sliced ​​tissue pieces of 0.5cm × 0.5cm wide, and placed in 3ml of 4wt% paraformaldehyde and 2.5wt % glutaraldehyde in 0.01M phosphate buffer (pH 7.2) fixative solution, pump air for 2min, replace with new fixative solution, and fix at room temperature for 5h; soak 3 times with 0.01M phosphate buffer solution (pH 7.2), 30 minutes each time; into the protective agent with different concentration gradients, the sucrose concentration of the protective agent is 2wt%-6wt%-10wt%-12wt%-14wt%-16wt%, and the time for each level is ...

Embodiment 2

[0034] Embodiment 2, primary laticifer and secondary laticifer histochemical display of annual sapling bark after mechanical injury and jasmonic acid treatment

[0035] In this embodiment, the bark of the sapling bark of rubber tree variety CATAS7-33-97 was mechanically injured and treated with jasmonic acid for 7 days to perform histochemical display of laticiferous ducts.

[0036] 1. Experimental conditions: select the saplings of the rubber tree variety CATAS7-33-97 in the stable stage, scrape off the epidermis and part of the cortex (length 1cm × width 1cm) at a place about 2cm above the base of the new shoot, and expose the injured part directly to the air After 7 days of exposure, cut the bark at the scratched part as bark material for mechanical damage treatment; apply lanolin containing 3mM methyl jasmonate to the injured part, and then cover it with a black plastic film. After 7 days of treatment, cut the bark at the scratched part as Jasmonate-treated bark material. ...

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Abstract

The invention discloses a histochemistry method capable of fast displaying a latex vessel of a bark of a rubber tree. The histochemistry method comprises the following steps: a) copping the bark of the rubber tree into section tissue blocks with the dimension of 0.5cm (length) * 0.5cm (width) and placing the section tissue blocks into a centrifugal tube containing stationary liquid; b) opening a lid of the centrifugal tube, placing the centrifugal tube into a glass dryer connected with a vacuum pump and exhausting air for 1-2min; c) exchanging fresh stationary liquid and rinsing for 3 times by buffer solution after fixing for 4-6h at room temperature, wherein the rinsing lasts 30min at each time; d) transiting the material in the step c) to protective agents with different concentration gradients from the stationary liquid; e) embedding the material in the protective agent with the highest concentration in the step d) into an OCT (Ornithine Carbamyl Transferase) embedding agent, placing the material on a refrigeration platform at -20 DEG C and condensing for 20min; f) taking out the material in step e) and sectioning the material by a conventional frozen section method; g) sticking sections by a common glass slide at room temperature; h) mounting the sections with a mounting medium and observing the sections by a fluorescence microscope. The histochemistry method disclosed by the invention has the advantages that the operation is simple, labor and land are saved, the repeatability and the universality are good, and batch screening can be realized.

Description

technical field [0001] The invention belongs to the technical field of agricultural histochemistry and relates to a method for displaying plant tissues, in particular to a histochemical method for quickly displaying latex ducts of rubber tree bark. Background technique [0002] There are two types of laticiferous ducts in rubber trees: primary laticiferous ducts formed by differentiation of primary meristem and secondary laticiferous ducts formed by differentiation of vascular cambium. Primary laticiferous ducts are distributed in primary structures of rubber tree leaves, flowers, fruits, seeds, roots and stems. The secondary milk ducts are distributed in the secondary phloem. The milk ducts directly related to natural rubber production are secondary milk ducts distributed in the bark of the trunk. There is evidence that the number of secondary laticifers in trunk bark is highly positively correlated with natural rubber production (Hao B Z, Wu JL. Biology of laticifers in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N21/64
Inventor 张世鑫田维敏
Owner RUBBER RES INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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