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Kit and method for marking microfilament cytoskeleton of pollen tube after apple pollination

A technology of cytoskeleton and pollen tube, applied in the field of plant biology, can solve the problems of complex composition of woody plant cell wall, difficulty in research on microfilament skeleton of pollen tube, difficulty in material collection, etc.

Active Publication Date: 2018-12-21
CHINA AGRI UNIV
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  • Application Information

AI Technical Summary

Problems solved by technology

It is more difficult to study the microfilament skeleton of the pollen tube in the apple style due to the difficulty in collecting materials in the pollen tube after pollination and the complex composition of the cell wall of woody plants

Method used

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  • Kit and method for marking microfilament cytoskeleton of pollen tube after apple pollination

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Embodiment

[0068] Example: Labeling the pollen tube microfilament cytoskeleton of apple after self- or cross-pollination

[0069] Since a large number of pollinated styles of apple 'Golden Delicious' were obtained and preserved in the early stage of this laboratory, styles in good condition were selected as research objects to mark the microfilament skeleton of pollen tubes in pollinated styles.

[0070] The brief description process is as follows: artificial pollination—fixing style—washing style—staining style with a mixed solution of microfilament dye reagent C and callose immunofluorescence reagent D—observing with laser confocal microscope.

[0071] Therefore, experiments were designed from the above steps to determine the method for labeling the microfilament cytoskeleton of pollen tubes in the style of apple after pollination.

[0072] 1. Marking the microfilament cytoskeleton of pollen tubes in apple post-pollination styles

[0073] 1. Collection of pollinated styles

[0074] O...

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Abstract

The invention belongs to the field of plant biotechnology and relates to a kit and method for marking a microfilament cytoskeleton of a pollen tube after apple pollination. The invention firstly provides a kit for marking the microfilament cytoskeleton of the pollen tube after the apple pollination, comprising a style fixer reagent A, a style washing liquid reagent B, a microfilament dye reagent Cand a callose immunofluorescence reagent D. Then the kit is applied for marking the microfilament cytoskeleton according to the following steps: 1) taking a pollinated style; 2) fixing the style by adopting the style fixer reagent A; 3) washing the style by adopting the style washing liquid reagent B; 4) dyeing the style by adopting mixed solution of the microfilament dye reagent C and the callose immunofluorescence reagent D; and 5) dropwise adding a fluorescence-quenching-resistant agent to the style, and observing the microfilament cytoskeleton of the pollen tube in the style under a laserscanning confocal microscope. The kit and the method which are provided by the invention realize rapid, sensitive, accurate and simple observation on a structure and an arrangement mode of the microfilament cytoskeleton in an apple pollen tube.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and relates to a kit and method for marking the cytoskeleton of pollen tube microfilaments in apples after pollination, in particular to a method for marking the cytoskeleton of pollen tube microfilaments in the styles of apples after self-flowering and cross-pollination Kits and methods. Background technique [0002] The plant cytoskeleton includes microfilaments and microtubules, which form different arrays and participate in many physiological processes such as cell division, growth and differentiation. The arrangement of the cytoskeleton in different eukaryotes is very different, and the actin cytoskeleton in the pollen tube cells plays an important role in the reproductive process. Previous studies mainly focused on cultivating pollen tubes in vitro and observing the morphology of the microfilament skeleton of the cells; however, the ability to clearly and intuitively observe the microfil...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N21/64
CPCG01N21/6428G01N33/533
Inventor 李天忠杨清顾钊宇李威李洋于杰刘春生
Owner CHINA AGRI UNIV
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