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Observation of mitochondrial DNA by mtg-dapi double staining in semi-thin slices of cucumber pollen

A semi-thin section and cucumber pollen technology, applied in the biological field, can solve problems such as no cytological evidence, and achieve a strong scientific effect

Active Publication Date: 2016-05-04
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

Therefore, it can be speculated that the germ cells during the development of cucumber pollen contain mitochondrial genomes and are passed on to offspring through fertilization, but there is no cytological evidence so far

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  • Observation of mitochondrial DNA by mtg-dapi double staining in semi-thin slices of cucumber pollen
  • Observation of mitochondrial DNA by mtg-dapi double staining in semi-thin slices of cucumber pollen
  • Observation of mitochondrial DNA by mtg-dapi double staining in semi-thin slices of cucumber pollen

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Embodiment Construction

[0043] MTG, as a mitochondrial membrane-specific fluorescent dye, is applied to the staining of mitochondria in living cells and fixed cell suspensions. This invention applies MTG to the fluorescent staining of mitochondria in resin semi-thin sections for the first time, which expands the application of MTG and can be used for other materials. Relevant research provides the basis. As we all know, most of the inheritance of plant organelles is maternal inheritance, that is, mitochondria follow the mother cell and enter the fertilized egg. The relevant research of cucumber shows that its mitochondrial gene is inherited from the paternal line, that is, the mitochondria follow the sperm into the fertilized egg, but it has not been verified by cytological observation. The present invention observes the mitochondria in the pollen sperm cells of the cucumber adherence stage by double fluorescent staining, thereby confirming that the mitochondrial genome is scattered around the reprod...

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Abstract

The invention discloses a method for performing MTG (Mito Tracker Green) and DAPI bifluorescence staining observation on resin semi-thin sections of cucumber pollen and relates to plant tissue fixation, plant anther Technovit7100 resin semi-thin sections and MTG-DAPI bifluorescence staining, belonging to the technical field of biology. The mitochondria and contained DNA in pollen reproductive cells in the cucumber adherence period are observed at the first time, the method can be used for exploring the change rule of mitochondria number in the cucumber pollen development process, and a foundation is laid for further researching related mechanisms of cucumber mitochondria paternal descent. The method also can be applied to mitochondria observation in other plant materials and is used for determining whether the plant materials have mitochondria genome paternal descent or biparental inheritance potentials.

Description

1. Technical field [0001] The invention relates to a method for performing MTG (MitoTrackerGreen) and DAPI double-staining fluorescence observation on resin semi-thin sections of cucumber pollen, relates to plant tissue fixation, plant anther Technovit7100 resin semi-thin section technology and fluorescent dyeing, and belongs to the field of biotechnology. 2. Background technology [0002] MTG (MitoTrackerGreen) is a mitochondrial (mitochondria) specific green fluorescent probe, the maximum excitation wavelength for detection is 488nm, and the maximum emission wavelength is 516nm. The staining of mitochondria by MTG does not depend on the mitochondrial membrane potential and can be used for staining of live cell homogenates and fixed cell suspensions. There have been no studies applying MTG for fluorescent staining of mitochondria in resin semi-thin sections of cells. [0003] DAPI is 4', 6-diamidino-2-phenylindole (4', 6-diamidino-2-phenylindole) is a very good DNA dye, wh...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/36G01N1/30
Inventor 陈劲枫赵娟沈佳娄群峰李季
Owner NANJING AGRICULTURAL UNIVERSITY
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