Fluorescence labeling method for free calcium ions of cotton pollen tube

A technology for fluorescent labeling and pollen tubes, which is applied in the fields of fluorescence/phosphorescence, preparation of test samples, and material excitation analysis, etc. The effect of excellent resolution

Inactive Publication Date: 2012-05-23
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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Problems solved by technology

However, the use of protoplasts as materials cannot truly reflect the changes in plant cells, and the requirements for preparing active protoplasts are harsh.
So far, due to the limitations of experimental materials and detection methods, effective methods and techniques for labeling free calcium ions in upland cotton powder tubes have not been reported

Method used

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  • Fluorescence labeling method for free calcium ions of cotton pollen tube
  • Fluorescence labeling method for free calcium ions of cotton pollen tube
  • Fluorescence labeling method for free calcium ions of cotton pollen tube

Examples

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Embodiment 1

[0038] Example 1. Effects of Nitrogen Ions Implanted into Sumian 22 Pollen on Calcium Ion Concentration in Germinated Pollen Tubes

[0039] First use the method of the present invention to inject the free Ca of the pollen tube to its germinated pollen tube after the nitrogen ion is implanted into the pollen of Sumian 22 (species for production). 2+ Perform fluorescent labeling, including the following steps:

[0040] 1) Nitrogen ion implantation

[0041] The ion beam implanter is an LCD-1000 multifunctional ion implanter made by the Institute of Plasma Physics, Chinese Academy of Sciences, and the ion source is nitrogen ions.

[0042] At about 16:30 on a sunny day in August every year, the flower buds that will open the next day are picked and spread on the table at room temperature. At about 9:00 in the morning of the next day, mix the pollen on the stamens of the open flowers and evenly scatter them in a glass petri dish with a diameter of 9 cm (to avoid the difference in ...

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Abstract

The invention discloses a fluorescence labeling method for free calcium ions of a cotton pollen tube, and belongs to the technical field of biology. The fluorescence labeling method comprises the following steps: perforating glass slide to make a small chamber for culturing the pollen tube; culturing pollen in the small chamber, and sprouting the pollen tube; loading Fluo-3AM into the cultured pollen tube by a low temperature incubation method; and observing dynamic variation of Ca2+ in the pollen tube under a confocal laser scanning microscopy. The invention establishes a method for researching the concentration change of the Ca2+ in the cotton pollen tube by using the confocal laser microtechnique, has simple operation and good labeling effect, plays an important role in the growth of the cotton pollen tube and fertilization reproduction research, and has important practical application value.

Description

1. Technical field [0001] The invention relates to a fluorescent labeling method for pollen tube free calcium ions in the field of plant biotechnology, and belongs to the field of biotechnology. 2. Background technology [0002] Intracellular free calcium ions are the second messenger for cells to feel stimuli, and its concentration can change rapidly in response to external environmental stimuli, thereby causing calcium ion downstream target proteins such as CaM (calmodulin), CDPK (CaM-dependent protein kinase ) and other activities, regulate many physiological and biochemical reactions in cells, thereby transducing the extracellular stimuli sensed by cells to adapt to the environment. Pollen germination and pollen tube elongation are a complex physiological and biochemical reaction process. This system has been increasingly used to study the molecular regulation mechanism of cell growth. Many research results in recent years have proved that free calcium in cells Ions are...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G01N1/28
Inventor 岳洁瑜唐灿明
Owner NANJING AGRICULTURAL UNIVERSITY
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