A method for marking apple pollen tube filaments

Abstract

The invention discloses a labeling method of an apple pollen tube microfilament. The labeling method comprises the following steps: cultivating apple pollen, curing the apple pollen, performing enzyme treatment on the apple pollen, treating the apple pollen with a surfactant, performing dye labeling, and performing scanning and imaging. The labeling method has the beneficial effect that the labeling method of gymnosperm pollen tube microfilament is referred and improved on the basis of the labeling method of pear pollen tube microfilament, dyes enter cells easier through curing, enzymolysis of cell walls and the surfactant, the interference of cell outer wall autofluorescence due to insufficient enzymolysis as TRITC-Phalloidin is applied are avoided when FITC-Phalloidin labeling is applied, so that relatively complete apple pollen tube microfilament can be stored; moreover, the labeling method is efficient and quick, can be used for obtaining relatively clear and bright pictures, thus filling up the blank of the apple pollen tube microfilament labeling, providing reference for labeling microfilament in rosaceae and angiosperm pollen tube, and achieving a high application value.

Description

technical field [0001] The invention relates to a method for marking apple pollen tube filaments. Background technique [0002] At present, there are almost no reports on the labeling of microfilaments in apple pollen tubes at home and abroad. The microfilament labeling technology in pollen mainly includes chemical staining methods: labeling phalloidin with fluorescent dyes, followed by freezing fixation combined with microinjection and using gene recombination technology to transfer the green fluorescent protein gene into living pollen tubes However, at present, the method of chemical staining is more simple and efficient. In recent years, there are relatively many studies on microfilaments in pollen of gymnosperms Pinaceae, and most of them are labeled with Rhodamine-Phalloidin. In angiosperm Rosaceae, the labeling of pear pollen tube microfilaments has been reported. During the labeling process, the steps of enzymatic hydrolysis of pollen tube wall and Triton decontamin...

AI Technical Summary

Problems solved by technology

In angiosperm Rosaceae, the labeling of pear pollen tube microfilaments has been reported. During the labeling process, the steps of enzymatic hydrolysis of pollen tube wall and Triton decontamination and permeabilization may be the reasons for the poor labeling effect.

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