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Method for quickly separating germ cells of angiosperms

An angiosperm and germ cell technology, applied in plant cells and other directions, can solve the problems of different lengths of pollen germination, germ cell membrane damage, and the effect of separation is uncertain, and achieves great practical application value, germ cell membrane integrity, and short separation time. Effect

Inactive Publication Date: 2014-04-30
BEIJING UNIV OF AGRI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The length of pollen germination is different and difficult to grasp, so the separation effect is uncertain, and the use of enzymes in the enzymatic hydrolysis method may cause damage to the isolated germ cell membrane, thereby affecting subsequent research or cultivation, so it is necessary to find a more suitable and simple method. Easy, easy-to-master separation method

Method used

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  • Method for quickly separating germ cells of angiosperms
  • Method for quickly separating germ cells of angiosperms
  • Method for quickly separating germ cells of angiosperms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1, separation tobacco germ cell

[0021] Include the following steps:

[0022] 1) Take fresh pollen from W38 tobacco;

[0023] 2) Prepare 13% mannitol solution;

[0024] 3) Put the pollen grains of step 1) in the solution of step 2) at 25°C for 45 minutes, the mass ratio of pollen grains to mannitol solution is 1:5;

[0025] 4) Microscopic examination: Microscopic examination with a microscope in bright field, the results are as follows figure 1 shown. The germ cell separation rate was as high as 93%, and the germ cell membrane was intact. It shows that the method can quickly and effectively separate the germ cells of tobacco.

[0026] Immediately released germ cells are spindle-shaped with two distinct tails. Over time, germ cells gradually become spherical, which undergoes intermediate states such as gradually shortening of the tail and germ cells becoming cylindrical. The length of time it takes for germ cells to change from elongated to spher...

Embodiment 2

[0027] Embodiment 2, separation iris germ cell

[0028] Include the following steps:

[0029] 1) Take the fresh pollen of iris;

[0030] 2) Prepare 13% mannitol solution;

[0031] 3) Put the pollen grains of step 1) in the solution of step 2) at 25°C for 15 minutes, the mass ratio of pollen grains to mannitol solution is 1:8;

[0032] 4) Microscopic examination: Microscopic examination with a microscope in bright field, the results are as follows figure 2 shown. The germ cell separation rate was as high as 92%, and the germ cell membrane was intact. It shows that this method can quickly and effectively isolate the germ cells of Iris.

Embodiment 3

[0033] Embodiment 3, separation broad bean germ cell

[0034] Include the following steps:

[0035] 1) Take fresh pollen from broad beans;

[0036] 2) Prepare 13% mannitol solution;

[0037] 3) Put the pollen grains of step 1) in the solution of step 2) at 25°C for 15 minutes, the mass ratio of pollen grains to mannitol solution is 1:10;

[0038] 4) Microscopic examination: Microscopic examination with a microscope in bright field, the results are as follows image 3 shown. The separation rate of germ cells is as high as 92%, and active germ cells account for more than 90%, and the germ cell membrane is complete. It shows that the method can quickly and effectively isolate the germ cells of faba bean.

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Abstract

The invention provides a method for quickly separating germ cells of angiosperms. The pollen of the angiosperms is put in mannitol solutions for a period of time for releasing to obtain the germ cells. The method for separating the germ cells of the angiosperms is short in separation time and simple in steps, plenty of active germ cells of the angiosperms can be obtained, germ cell membranes are complete, and the method has a large practical application value.

Description

technical field [0001] The invention relates to a method for rapidly separating angiosperm germ cells. Background technique [0002] Germ cells are the precursors of sperm cells, which further divide to form a pair of sperm cells. In the two-cell pollen types that make up the majority of angiosperms, the pollen contains only germ cells and vegetative cells when mature. Since germ cells and sperm cells are embedded in pollen tubes or pollen grains, they need to be separated for further manipulation and research on active germ cells. [0003] From the initial grinding method to the osmotic pressure shock method and then to the enzymatic hydrolysis method, the separation of germ cells has gone through a separation process from complex to simple, from a small amount to large-scale, and the separation methods of germ cells with different characteristics are also different. Currently, the most conventional method for separating germ cells is the two-step osmotic shock method, wh...

Claims

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Application Information

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IPC IPC(8): C12N5/04
Inventor 房克凤秦岭郝敬虹杨瑞杨柳王建立张卿于同泉
Owner BEIJING UNIV OF AGRI
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