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Method for inducing corn haploids

A technique of haploid and haploid induction, applied in chemical instruments and methods, botanical equipment and methods, applications, etc., can solve the problems of low commercial application value, inconvenient operation, and high production cost

Inactive Publication Date: 2015-02-11
CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Applying the principle of plant cell totipotency mainly uses tissue culture technology to induce female gametes or male gametes. Commonly used are anther culture and microspore culture, but this method is inconvenient to operate and the production cost is high. In addition, in some species It is difficult to produce haploids using in vitro culture, including corn, so the commercial application value is not high

Method used

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  • Method for inducing corn haploids
  • Method for inducing corn haploids
  • Method for inducing corn haploids

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Example 1. Cultivation of maize haploid inducer line CAU5 with higher maize haploid inducer rate than CAU5 YFP

[0043] 1. Introduce the coding gene of the centromere-specific histone mutant into the first-generation plant of the cross between the maize inbred line HiIIA (maternal parent) and the maize inbred line HiIIB (male parent) to obtain the centromere-specific Transgenic corn plant HiII with histone mutant YFP

[0044] The recipient of the transgene is the hybrid F1 generation of inbred lines HiIIA and HiIIB. The gene encoding the centromere-specific histone mutant was introduced into the maize inbred line HiIIA and the maize inbred line HiIIB through the expression vector pZY101-35S-CENH3-YFP, which is the expression vector of the centromere-specific histone mutant CENH3-YFP encoding gene. First generation (F1) plants. pZY101-35S-CENH3-YFP comes from the Jiming Jiang laboratory (Weiwei Jin, Jiming Jiang.Histone modifications associated with both A and B chromosomes...

Embodiment 2

[0080] Example 2. Maize haploid induction line CAU5 YFP Induction of maize haploid

[0081] The experiment adopted a random block design in the field. The experiment included 2 treatments: corn haploid induction line CAU5 YFP Treatment and corn haploid induction line CAU5 treatment, the experimental method is as follows:

[0082] Maize haploid induction line CAU5 YFP The experimental method of treatment is as follows: from the corn haploid induction line CAU5 of Example 1 YFP Randomly select 24 individual plants, mix the pollen of the 24 individual plants, and then pollinate 10 Zhengdan 958 (female parents). Cross in the field to induce maize haploids, harvest the hybrid seeds, and detect the haploids of the hybrid seeds Count, calculate the corn haploid induced line CAU5 YFP The haploid induction rate of Zhengdan 958 showed that a total of 2640 hybrid seeds were harvested, of which 312 were haploid, and the corn haploid induction line CAU5 YFP The haploid induction rate of Zhengdan...

Embodiment 3

[0087] Example 3. Identification of corn haploid inducible line CAU5 by observing seed fluorescence YFP Induced maize haploid

[0088] Example 1 Cultivated corn haploid induction line CAU5 YFP Not only can it be used as a haploid induction line to produce haploids in the offspring, but it can also stably express the fluorescent fusion protein CENH3-YFP and locate it at the centromere position, such as figure 2 . By directly observing the fluorescence signal of the seeds in a stereoscopic fluorescence microscope, it can also be seen that the corn haploid induction line CAU5 YFP Fluorescence difference between CAU5 seeds and corn haploid induction line, such as Figure 4 , The two kernels on the left are corn haploid induced line CAU5 YFP The two grains on the right are the grains of the corn haploid inducer line CAU5, and the corn haploid inducer line CAU5 YFP It shows a more obvious YFP signal, and the corn haploid inducible line CAU5 can also show a certain degree of autofluoresc...

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Abstract

The invention discloses a method for inducing corn haploids. The method comprises the following steps: pollinating a female parent by a male parent corn haploid induction line with a higher corn haploid inductivity than a corn haploid induction line CAU5, and harvesting hybridized F1 generation grains to obtain corn haploids. A method for realizing the corn haploid induction line comprises the following steps: 1, introducing coding gene of a specific centromere histone mutant into a hybrid first generation plant of a corn inbred line HiIIA and a corn inbred line HiIIB to obtain a transgenic corn plant; and 2, hybridizing the specific histone mutant as a female parent with the CAU5 to obtain an F1 generation, hybridizing the F1 generation as a female parent with the CAU5 to obtain a BC1F1 generation, hybridizing the BC1F1 generation as a female parent with the CAU5 to obtain a BC2F1 generation, and continuously selfing the BC2F1 generation at least five generations to obtain the corn haploid induction line.

Description

Technical field [0001] The present invention relates to a method for inducing corn haploids. Background technique [0002] Haploid refers to an individual with the number of gamete chromosomes. Since the number of chromosomes of a haploid is only the number of chromosomes of the gametophyte, once it is doubled, a pure line can be obtained, and pure line breeding is a very key link in the utilization of heterosis, so the utilization value of haploid is undoubtedly huge. [0003] Haplotype refers to the way to obtain haploids through various methods, obtain homozygous diploids through chromosome doubling, and select at the haploid and diploid levels in order to breed excellent pure line varieties. In 1964, Datura anther culture successfully obtained haploid plants (Guha and Maheshwari, 1964), followed by an upsurge in haploid research, and haploid research has developed rapidly. The advantages of haplotypes are as follows: 1) Shorten the breeding process. In conventional breeding, ...

Claims

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Application Information

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IPC IPC(8): A01H1/02A01H1/04C07K19/00
Inventor 金危危陈绍江孙占勇朱奇琳徐小炜赵鑫徐丽
Owner CHINA AGRI UNIV
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