Mitochondrial fluorescence labeling method for angiosperm flower organs

An angiosperm, fluorescent labeling technology, applied in fluorescence/phosphorescence, preparation of test samples, material excitation analysis, etc., can solve the problem of lagging research on mitochondria of flower organs, and achieve the effect of good labeling effect and high application value.

Inactive Publication Date: 2014-08-13
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, fluorescent labeling of mitochondria in floral organs of non-model organisms has not been reported, which has led to a lag in the study of mitochondria in floral organs

Method used

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  • Mitochondrial fluorescence labeling method for angiosperm flower organs
  • Mitochondrial fluorescence labeling method for angiosperm flower organs
  • Mitochondrial fluorescence labeling method for angiosperm flower organs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, the fluorescent labeling method of the tepal mitochondria of Magnolia Magnolia

[0037] 1. Fluorescent labeling of the mitochondria of the tepals of Magnolia magnolia

[0038] The buffer solution containing 100nM Mitotracker Red CMXRos consists of Mitotracker Red CMXRos, sucrose, boric acid and water, the final concentration of Mitotracker Red CMXRos in the buffer solution is 100nM, and the mass percentage of sucrose in the buffer solution is 2% , the mass percentage of boric acid in the buffer solution is 0.01%, and the pH value of the buffer solution is 6.5.

[0039] 1) Take a fresh perianth piece, and peel off 4mm of epidermal monolayer cells on the perianth piece with razor blade tweezers 2 , if it is not a monolayer of cells, lightly scrape the epidermis with a blade until the epidermis on the perianth slices appears transparent, and obtain the treated perianth slices.

[0040] 2) Stain the treated perianth slices in a buffer solution containing 100...

Embodiment 2

[0045] Example 2, Magnolia style mitochondrial fluorescence staining and microscope observation

[0046] 1. Fluorescence labeling of magnolia styloid mitochondria

[0047] The buffer solution containing 100nM Mitotracker Red CMXRos consists of Mitotracker Red CMXRos, sucrose, boric acid and water, the final concentration of Mitotracker Red CMXRos in the buffer solution is 100nM, and the mass percentage of sucrose in the buffer solution is 2% , the mass percentage of boric acid in the buffer solution is 0.01%, and the pH value of the buffer solution is 7.0.

[0048]1) Take a fresh magnolia style, cut the style longitudinally with a blade, and then stick the style on the stage of a Leica VT12000S oscillating microtome with 502 glue. The blade of the slicer and the style are kept at an angle of 30°, and the slice thickness parameter is set to 80 μm. Get the treated style.

[0049] 2) The treated style was stained in a buffer solution containing 100 nM Mitotracker Red CMXRos, an...

Embodiment 3

[0054] Example 3, Rhododendron style mitochondrial fluorescence staining and microscope observation

[0055] The buffer solution containing 200nM Mitotracker Red CMXRos consists of Mitotracker Red CMXRos, sucrose, boric acid and water, the final concentration of Mitotracker Red CMXRos in the buffer solution is 100nM, and the mass percentage of sucrose in the buffer solution is 2% , the mass percentage of boric acid in the buffer solution is 0.01%.

[0056] 1) Take a fresh rhododendron style, cut the style longitudinally with a blade, and obtain a longitudinal section of the style with a slice thickness parameter of 80 μm with a single layer of cells on the edge, which is the treated style.

[0057] 2) The treated style was stained in a buffer solution containing 200 nM Mitotracker Red CMXRos, and stained at 25° C. for 20 min to obtain a stained style. 2. Detection

[0058] The stained style was washed three times with a buffer solution composed of 2% sucrose and 0.01% boric ...

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Abstract

The invention discloses a mitochondrial fluorescence labeling method for angiosperm flower organs. The method provided by the invention comprises the following steps: dyeing the treated angiosperm flower organs in a buffer solution containing Mitotracker Red CMXRos, thereby realizing fluorescence labeling of mitochondria of the angiosperm flower organs. An experiment proves that the mitochondrial fluorescence labeling method for angiosperm flower tepals has the advantages that a labeling effect is good, and uniform mitochondrion distribution around cells can be observed under a laser scanning confocal microscope; after being fixed by a fixing solution, a slide labeled by Mitotracker can be made into a permanent slide to be observed; mitochondria in the flower organs can be labeled simply, rapidly and quickly, a foundation is laid for distribution and displacement of the mitochondria of the flower organs, and the actual application value is high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a mitochondrial fluorescent labeling method for flower organs of angiosperms. Background technique [0002] As the reproductive organs of plants, flowers are composed of sepals, petals, stamens and pistils, and are responsible for the entire process of plant pollination, fertilization and fruiting, so they have become the focus of botany research. In response to changes in the external environment, plants have evolved a variety of response mechanisms, such as plant flowering heat generation, self-incompatibility, and styles that guide tissue secretion substances to guide the growth of pollen tubes, etc., which play an important role in ensuring the reproductive process of plants. important role. As an important organelle for energy metabolism, mitochondria can move and move in response to cellular events. The flower organ is an important place for reproduction and growth, with stro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30G01N21/64
Inventor 王若涵张出兰张志翔纪翔宇刘翔宇
Owner BEIJING FORESTRY UNIVERSITY
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