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Doubling method for plant haploid

A haploid and plant technology, applied in the biological field, can solve the problems of low doubling rate, complicated operation, and inability to realize the large-scale breeding of corn haploid breeding, and achieve high safety for people and the environment, simple operation, and large-scale promotion The effect of the process

Inactive Publication Date: 2016-09-07
SAAS BIOTECH & NUCLEAR TECH RES INST
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

However, the haploid breeding technology also has disadvantages, that is, the doubling rate in maize haploid breeding is low, and the operation is often complicated, and it is impossible to achieve large-scale maize haploid breeding

Method used

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  • Doubling method for plant haploid
  • Doubling method for plant haploid

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Present embodiment provides a kind of corn haploid doubling method, comprises the following steps:

[0020] 1) Disinfect the haploid corn kernels and place them in fine sand at 26-30°C to moisturize and accelerate germination;

[0021] 2) When the radicle length is 1-1.5cm, the young shoots are taken out, and placed at 4°C for 8-12 hours in the dark;

[0022] When the root tip length of corn is 1-1.5cm, its mitosis is in the most active period. Roots less than 1cm or greater than 1.5cm can also be used, but the effect is second. Therefore, the present embodiment selects the radicle length as 1-1.5cm.

[0023] The main function of this step is to reduce the cell division speed of the corn root tip, so that most of the cells are in the metaphase of mitosis, so that more diploid cells can be obtained in the subsequent colchicine treatment, and the doubling rate can be increased.

[0024] 4℃ is the critical temperature for maize radicle growth and meristematic zone mitosi...

Embodiment 2

[0035] 1) Germination of corn haploid seeds: sterilize the haploid seeds and soak them in distilled water for 10 hours; insert the soaked seeds evenly into a germination tray filled with sand with a humidity of about 75% (thickness of the sand tray is more than 10cm) , spread a wet towel, and place it in an incubator at 26°C for 2 days in the dark, and replenish water in time.

[0036] Excision of the top of the radicle: when the haploid radicle of the corn grows to about 1.5 cm, cut off the top 1-2 mm of the radicle of the haploid young germ of the corn, and then cut 1-2 mm longitudinally along the direction of the radicle.

[0037] 2) Colchicine solution preparation: prepare colchicine solutions with 3 different concentrations (mass percentage): 0.02% colchicine+2% dimethyl sulfoxide; 0.04% colchicine+2% dimethyl sulfoxide Sulfone; 0.049% colchicine + 2% dimethyl sulfoxide; 0.05% colchicine + 2% dimethyl sulfoxide;

[0038] 3) Sprout soaking: the radicle of the radicle trea...

Embodiment 3

[0042] This method is currently the most common method - the germination seed treatment method

[0043] Colchicine concentration: 0.06%, 0.08%, 0.1%

[0044] Germination of haploid seeds: sterilize the haploid seeds and soak them in distilled water for 10 hours; place the seeds evenly in a germination tray covered with a wet towel, and place them in a suitable environment at 26°C for about 24 hours in dark light. Take out after the radicle breaks.

[0045] Treatment: Put the seed embryos of Lubai (radical root breach) face down in the germination tray, and then add appropriate amount of colchicine solutions of different concentrations mentioned above, whichever is just submerged the seed embryo surface. The treatment time is 4h, 8 hours and 12 hours respectively. The processing temperature is normal temperature (about 22 degrees).

[0046]The treated seeds were washed with running water for about 40 minutes, sown in a nutrient bowl, and moved into the field at the stage of ...

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Abstract

The invention provides a doubling method for a plant haploid. The method includes the steps that the radicle part of a treated haploid bud is soaked in mixed liquid of colchicines with the mass percentage of 0.02% to 0.05% and dimethyl sulfoxide with the mass percentage of 2%, wherein the treatment time ranges from 18 h to 36 h, and the temperature ranges from 14 DEG C to 18 DEG C. By means of the haploid doubling method, the technical problem that the doubling rate is low in haploid breeding is solved, the method is easy to operate and higher in safety to people and the environment, and the scalization process of haploid breeding is effectively promoted.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a plant haploid doubling method. Background technique [0002] With the increase of population and the development of economy, the demand of corn keeps rising. The reduction of arable land and the deterioration of the environment have made people's demand for new varieties of maize with high yield, high quality and strong stress resistance increasingly urgent. However, it takes a lot of manpower and time to obtain homozygous inbred lines with high combining ability by conventional breeding methods. Conventional breeding techniques have been increasingly unable to meet people's breeding needs. In recent years, the development of biotechnology methods such as haploid breeding technology, genetic engineering breeding, and molecular marker-assisted breeding has improved breeding efficiency and opened up new ways of maize breeding. [0003] The application of haploid technology in maiz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/08
CPCA01H1/08
Inventor 宋军杜文平余桂容陈谦徐利远林杨
Owner SAAS BIOTECH & NUCLEAR TECH RES INST
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