Method for doubling cone haploids

A haploid and corn technology, applied in botany equipment and methods, applications, plant genetic improvement, etc., can solve problems such as plant damage, low survival rate, and affecting root system growth, and achieve scale and engineering, and improve The effect of doubling efficiency and improving survival rate

Inactive Publication Date: 2012-09-12
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, colchicine is very toxic and can easily cause serious damage to seedlings, which can easily affect the doubling efficiency of haploids
In addition, overall, the doubling efficiency of the soaking method is not ideal
The main reason is that the seed soaking method is more serious to the plant damage, and the survival rate is lower; and, the bud soaking method used by the predecessors all soaks the whole seedling in the colchicine solution, not only the young shoots absorb the colchicine, but also the roots of the seedlings also absorb the colchicine. Absorbed a large amount of colchicine solution, which affected the growth of the root system and caused a large number of seedlings to die

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Maize haploid doubling method and control experiment (1)

[0033] The corn haploid doubling method provided in this embodiment consists of the following steps:

[0034] 1. Seed germination: Clean the haploid seeds, evenly place them in a germination tray covered with wet towels, place them in an incubator at 25°C for 4 days in the dark, and add water in time.

[0035] 2. Resection of the coleoptile tip: When the corn haploid shoots grow to 2 cm, the coleoptile tip of the corn haploid shoots is removed without damaging the true leaves and growth points, and the coleoptile tip of the corn haploid is obtained. bud.

[0036] 3. Soaking the young buds: Take the centrifuge tube, add 500μL 0.6mg / mL colchicine solution (with a final concentration of 2.0% dimethyl sulfoxide) into each centrifuge tube, and then remove the top of the coleoptile. Put the young shoots upside down into the centrifuge tube, so that the embryo part of the young shoots are soaked in the colchicine so...

Embodiment 2

[0042] Example 2 Maize haploid doubling method and control experiment (2)

[0043] The corn haploid doubling method provided in this embodiment consists of the following steps:

[0044] 1. Seed germination: Clean the haploid seeds, evenly place them in a germination tray covered with wet towels, and place them in an incubator at 20°C for 5 days in the dark, and add water in time.

[0045] 2. Resection of the coleoptile tip: When the corn haploid shoot grows to 3 cm, the coleoptile tip of the corn haploid shoot is removed without damaging the true leaf and growth point, and the corn haploid young with the tip of the coleoptile removed bud.

[0046] 3. Soaking the young shoots: Take the centrifuge tube, add 300μL 0.8mg / mL colchicine solution (with a final concentration of 1% dimethyl sulfoxide) into each centrifuge tube, and then remove the top of the coleoptile. Put the young shoots upside down into the centrifuge tube, so that the embryo part of the young shoots are soaked in the col...

Embodiment 3

[0052] Example 3 Maize haploid doubling method and control experiment (3)

[0053] The corn haploid doubling method provided in this embodiment consists of the following steps:

[0054] 1. Seed germination: Clean the haploid seeds, evenly place them in a germination tray covered with wet towels, and place them in an incubator at 28°C for 3 days in the dark, and add water in time.

[0055] 2. Resection of the coleoptile tip: When the corn haploid shoot grows to 3 cm, the coleoptile tip of the corn haploid shoot is removed without damaging the true leaf and growth point, and the corn haploid young with the tip of the coleoptile removed bud.

[0056] 3. Soaking young shoots: Take centrifuge tubes, add 500μL 0.4mg / mL colchicine solution (with a final concentration of 5% dimethyl sulfoxide) into each centrifuge tube, and then remove the top of the coleoptile. Put the young shoots upside down into the centrifuge tube, so that the embryo part of the young shoots are soaked in the colchicine...

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PUM

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Abstract

The invention provides a method for doubling cone haploids. The method comprises the following steps: 1, cutting off the tips of coleoptiles of buds of corn haploids; and 2, soaking embryos of the buds of the corn haploids without the tips of coleoptiles in mixed liquid of colchicines and dimethyl sulfoxide, so as to obtain corn buts with doubled chromosome. The method for doubling cone haploids, which is provided by the invention, can remarkably improve the survival rate and maturing rate of the corn haploids, wherein the survival rate is higher than 70%, which is far higher than the survival rate of a control group which is generally lower than 50%, the maturing rate is higher than 10%, which is far higher than the maturing rate of the control group which is generally about 5%; moreover, the use amount of colchicines is 5-6 times. When applied to a haploid breeding system, the method can effectively promote the reform of the corn breeding technology to realize large-scale and engineering haploid breeding.

Description

Technical field [0001] The present invention relates to the field of biotechnology, specifically to the field of agricultural breeding, and more specifically, to a corn haploid doubling technology with high efficiency and low consumption, which can significantly increase the doubling rate. Background technique [0002] The use of Doubled haploid (DH) in maize can shorten the breeding time of inbred lines and speed up the breeding process, which has become the main method of maize breeding. Obtaining enough haploids and doubling to obtain DH lines are the basic steps of DH breeding. However, natural doubling of haploids in maize is more difficult. At present, the successful doubling of haploids is a limiting factor for the large-scale application of haploid seed technology, especially the successful doubling of tassels. Haploid doubling is divided into natural doubling and drug doubling. The natural doubling rate of many materials is less than 5%, and some materials do not even o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/08
Inventor 段民孝赵久然刘新香王元东
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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