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Soybean sterile gene mutant, construction method and application in photosensitive fertility regulation

A construction method and soybean technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of long time and cumbersome process, and achieve the effect of reducing time, obvious economic value, and low precision requirements.

Active Publication Date: 2022-04-29
JILIN ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is of great significance to provide as many nuclear male sterile lines with different genetic composition as possible to increase the diversity of soybean varieties of sterile lines and to provide more optional resources for hybrid breeding and intelligent breeding. At the same time, the existing nuclear male sterile lines The method of male sterile lines mainly relies on screening, which is cumbersome and time-consuming. It is urgent to provide a method that can quickly and accurately obtain the male sterile line of soybean varieties. sexual regulation

Method used

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  • Soybean sterile gene mutant, construction method and application in photosensitive fertility regulation
  • Soybean sterile gene mutant, construction method and application in photosensitive fertility regulation
  • Soybean sterile gene mutant, construction method and application in photosensitive fertility regulation

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Experimental program
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Embodiment 1

[0062] In this embodiment, the sterile mutant strain obtained from the Changchun test base is used as a material, and a soybean sterile gene mutant is provided by gene mapping method, the nucleotide sequence of which is shown in SEQ ID No. The difference in gene sequence is that the amino acid residue encoded by codons 10236294-10236296 of the wild-type ORF2 gene sequence is valine, while the amino acid residue encoded by nucleotides +1882-+1884 of the mutant in this example is is tryptophan, and the codon corresponding to nucleotides +1885 to +1887 of the mutant in this embodiment is the stop codon TAA.

[0063] The specific positioning method is as follows:

[0064] First, the results of mature period of the sterile plants in the Changchun Experimental Base of Jilin (43°50′N, from the first flowering stage to the final flowering stage: day and night time range 14:50′ / 9:10′-15:25′ / 8:35′) were studied. number of pods, the investigation was carried out and the results were as ...

Embodiment 2

[0070] This embodiment also provides a soybean sterility gene mutant, which differs from the embodiment 1 in that the amino acid residue encoded by nucleotides +1882-+1884 of the mutant in this embodiment is valine.

Embodiment 3

[0072] In order to verify the conclusion that the abnormal expression of the PHD-finger domain of the ORF2 gene of soybean chromosome 2 leads to the conclusion of fertility changes, this example uses CRISPR gene editing technology to construct the KNOCKOUT vector, and its two target sequences are respectively located in soybean 2. The positions 55bp-74bp and 64bp-83bp upstream of the PHD-finger domain of the chromosome ORF2 gene, and the KNOCKOUT vector was genetically transformed into the broad-adapted soybean variety Williams82, and T 1 transgenic plants.

[0073] The primer sequences targeting 55bp~74bp (primer pair 1) and 64bp~83bp (primer pair 2) are shown in Table 2:

[0074] Table 2 Primer sequences targeting 55bp~74bp and 64bp~83bp

[0075]

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Abstract

The invention relates to the technical field of agricultural breeding, in particular to a soybean sterile gene mutant, a construction method and application of the soybean sterile gene mutant in photosensitive fertility regulation. The soybean sterile gene mutant provided by the invention is located in a soybean chromosome 2, and an abnormality exists in a PHD-finger structural domain region of an ORF2 gene, so that compared with a wild type, the gene mutant has the advantages that the PHD-finger structural domain transcription is terminated in advance or the PHD-finger structural domain function is lost after smooth expression, thereby finally changing the fertility of the soybean and causing pollen abortion of the mutant. Through identification, the fertility of the soybean containing the mutant also shows photosensitivity, and the adjustment of the fertility of a plant transformed with the mutant soybean can be realized by adjusting the illumination length, so that a brand-new sterile line resource is provided for soybean hybrid seed production or sterile line seed reproduction; and the risk caused by failure of other sterile line resources due to a certain reason is avoided.

Description

technical field [0001] The invention relates to the technical field of agricultural breeding, in particular to a soybean sterile gene mutant, a construction method and an application in light-sensitive fertility regulation. Background technique [0002] Soybean (Glycine max(L.)Merr.) is an important economic crop in my country. However, there is a large gap between the actual production capacity of domestic soybean and the actual demand. Increasing the yield per unit area is the most important goal of soybean breeding in my country. As an effective means to increase soybean yield, hybrid soybean has created huge economic benefits and social value in production. As the core material of hybrid seed production, the CMS itself is sensitive to the environment often determines the difficulty of hybrid seed production, especially the CMS germplasm in the "two-line method", the conversion of fertility in a specific environment Ability is the decisive factor for the stability of hyb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82A01H5/02A01H6/54A01H1/02
CPCC07K14/415C12N15/8218C12N15/8237C12N15/8289A01H1/02
Inventor 张春宝侯晶晶范薇薇马瑞瑞赵丽梅孙连军
Owner JILIN ACAD OF AGRI SCI
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