Method for cultivating maize haploid induction line whose haploid induction rate is higher than that of maize haploid induction line cau5

A haploid induction system and haploid induction technology are applied in chemical instruments and methods, botanical equipment and methods, recombinant DNA technology, etc., which can solve the problem of low commercial application value, inconvenient operation, haploid Difficulty and other issues

Inactive Publication Date: 2017-08-25
CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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

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  • Method for cultivating maize haploid induction line whose haploid induction rate is higher than that of maize haploid induction line cau5
  • Method for cultivating maize haploid induction line whose haploid induction rate is higher than that of maize haploid induction line cau5
  • Method for cultivating maize haploid induction line whose haploid induction rate is higher than that of maize haploid induction line cau5

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1. Cultivation of the maize haploid induction line CAU5 with a higher maize haploid induction rate than the maize haploid induction line CAU5 YFP

[0056] 1. The gene encoding the centromere-specific histone mutant was introduced into the first-generation hybrid plants of the corn inbred line HiIIA (female parent) and the corn inbred line HiIIB (male parent) to obtain the centromere-specific histone mutant. Histone mutant transgenic maize plant HiII YFP

[0057] The recipient of the transgene was the hybrid F1 generation of the inbred lines HiIIIA and HiIIB. The gene encoding centromere-specific histone mutant was introduced into the hybridization of maize inbred line HiIIA and maize inbred line HiIIIB through the expression vector pZY101-35S-CENH3-YFP encoding gene of centromere-specific histone mutant CENH3-YFP First generation (F1) plants. pZY101-35S-CENH3-YFP comes from Jiming Jiang's lab (Weiwei Jin, Jiming Jiang. Histone modifications associated with b...

Embodiment 2

[0093] Embodiment 2, maize haploid induction line CAU5 YFP induced maize haploid

[0094] The experiment adopts random block design in the field, and the experiment includes 2 treatments: maize haploid induction line CAU5 YFP Treatment and maize haploid induction line CAU5 treatment, the experimental method is as follows:

[0095] Maize Haploid Inducer Line CAU5 YFP The experimental method of processing is as follows: from the maize haploid induction line CAU5 of embodiment 1 YFP Randomly select 24 individual plants, mix the pollen of 24 individual plants and pollinate 10 Zhengdan 958 (female parents), conduct hybridization in the field to induce maize haploid, harvest hybrid seeds, and detect the haploid of hybrid seeds Number, calculate the maize haploid induction line CAU5 YFP For the haploid induction rate of Zhengdan 958, the results showed that a total of 2640 hybrid seeds were harvested, of which 312 were haploid, and the maize haploid induction line CAU5 YFP The h...

Embodiment 3

[0100] Example 3. Identification of maize haploid inducible line CAU5 by observing seed fluorescence YFP induced maize haploid

[0101] The maize haploid induction line CAU5 cultivated in Example 1 YFP Not only can it be used as a haploid inducing line to produce haploid in the offspring, but it can also stably express the fluorescent fusion protein CENH3-YFP and locate at the centromere, such as figure 2 . By directly observing the fluorescent signal of seeds under a stereoscopic fluorescence microscope, it can also be seen that the maize haploid induction line CAU5 YFP Differences in fluorescence from seeds of the maize haploid inducible line CAU5, as Figure 4 , the left two grains are maize haploid induction line CAU5 YFP The two grains on the right are the grains of the corn haploid induction line CAU5, and the corn haploid induction line CAU5 YFP It showed a more obvious YFP signal, and the maize haploid induced line CAU5 also showed some autofluorescence, but ther...

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Abstract

The invention discloses a method for cultivating a maize haploid induction line whose haploid induction rate is higher than that of the maize haploid induction line CAU5. The method comprises 1) introducing the coding gene of the centromere-specific histone mutant into the first-generation hybrid plants of the corn inbred line HiIIA and the corn inbred line HiIIB, and obtaining the centromere-specific histone mutant containing Transgenic corn plants; 2) using the transgenic corn plants as the female parent to cross with the corn haploid induction line CAU5 to obtain an F1 generation, and using the F1 generation as the female parent to perform hybridization with the corn haploid induction line CAU5 Hybridize to obtain BC1F1 generation, and use the BC1F1 generation as the female parent to cross with the corn haploid induction line CAU5 to obtain the BC2F1 generation; the BC2F1 generation is continuously selfed for at least 5 generations, and the maize haploid induction rate is higher than A maize haploid inducer line of the maize haploid inducer line CAU5.

Description

technical field [0001] The invention relates to a method for cultivating a maize haploid induction line whose haploid induction rate is higher than that of the maize haploid induction line CAU5. Background technique [0002] Haploid refers to an individual with the number of gamete chromosomes. Since the number of haploid chromosomes is only the number of gametophyte chromosomes, once doubled, pure lines can be obtained, and pure line selection is a very critical link in the utilization of heterosis, so the utilization value of haploid is undoubtedly huge. [0003] Haploid breeding refers to the way of obtaining haploids by various methods, obtaining homozygous diploids through chromosome doubling, and selecting at the haploid and diploid levels in order to breed excellent pure-line varieties. In 1964, haploid plants were successfully obtained by cultured anthers of Datura datura (Guha and Maheshwari, 1964), and then there was an upsurge in haploid research, which developed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N15/62C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C07K19/00A01H5/00A01H1/02
Inventor 金危危陈绍江孙占勇徐小炜赵鑫朱奇琳
Owner CHINA AGRI UNIV
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