Male fertility maintaining method and application thereof

A male sterile line and fertility restorer gene technology, applied in the field of plant biology, can solve problems such as unstable fertility, narrow recovery sources, and difficult reproduction of sterile lines

Active Publication Date: 2017-10-20
BEIJING NEXT GENERATION HYBRID WHEAT BIOTECHNOLOGY CO LTD +1
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Problems solved by technology

The three-line method has not been widely used in production due to the difficulty in breeding sterile lines, narrow recovery sources, and cytoplasmic side effects.
At the same time, in order to solve the technical bottlenecks of the existing "three-line" and two-line" hybridization technologies of wheat, such as unstable sterile line fertility, limited hybrid variety resources, complicated seed production technology, and high seed production costs, it provides more many possibilities

Method used

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  • Male fertility maintaining method and application thereof
  • Male fertility maintaining method and application thereof
  • Male fertility maintaining method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Embodiment 1, the cloning of Thiopyrum elongatum FRG1 gene

[0094] The wheat Lanzhou nuclear male sterile mutant is a typical recessive mutation controlled by a single gene. The 4AgS-ms double-terminal heterogeneous addition line is the addition of half of the 4Ag chromosome (4AgS) of Echinopsis elongatus in the Lanzhou nuclear male sterility mutant (Zhou Kuanji et al., 1996). The additional 4AgS chromosome can restore the mutant male sterility. The phenotype of fertility indicated that the chromosome of E. elongatum 4AgS contained the fertility restoration gene Fertility Restoration gene1 (FRG1).

[0095] The length of the 4AgS chromosome in the 4AgS-ms double-terminal allosome addition line is significantly shorter than that of all wheat chromosomes, and its size is estimated to be about 1 / 4 of the longest wheat chromosome, about 250Mb. The 4AgS chromosome of Echinopsis elongatum was isolated by flow cytometry, and the 4AgS reference genome was assembled using the s...

Embodiment 2

[0099] Embodiment 2, the homologous gene of EeFRG1 in wheat

[0100] Using the EeFRG1 gene (SEQ ID NO: 2) to compare in the IWGSC (International Wheat GenomeSequencing Consortium) database, two homologous genes were found, named TaFRG1-1 and TaFRG1-2, and their genomic DNA sequences are shown in SEQ ID NO Shown in: 5 and SEQ ID NO: 8, the CDS sequence deduced according to the EeFRG1 gene is shown in SEQ ID NO: 6 and SEQ ID NO: 9 respectively, and the protein sequence is shown in SEQ ID NO: 7 and SEQ ID NO: 10 respectively .

[0101] Using Clustal Omega, the genomic DNA sequence, CDS sequence and protein sequence of the TaFRG1-1 gene were compared with the EeFRG1 gene, and the results are as follows Figure 1-3 As shown, the genomic DNA sequence identity of TaFRG1-1 gene and EeFRG1 gene is 88.5%, the identity of CDS sequence is 95.0%, and the identity of protein sequence is 95.9%.

[0102] Using Clustal Omega, the genomic DNA sequence, CDS sequence and protein sequence of the...

Embodiment 3

[0104] Example 3, Sequence Alignment of EeFRG1 Gene and Homologous Genes in Uraltu Wheat, Goat Grass, Sorghum, Corn, and Millet

[0105]The protein sequence of the EeFRG1 gene was searched in databases such as NCBI, and the homologous proteins predicted in the genomes of wheat Uraltu, goat grass, sorghum, maize, and millet were obtained. Among them, maize contains two homologous genes of EeFRG1, named respectively for ZmFRG1-1 and ZmFRG1-2. Wherein, the genomic DNA sequence of the fertility gene TuFRG1 in Uraltu wheat is shown in SEQ ID NO: 11, the CDS sequence is shown in SEQ ID NO: 12, and the protein sequence is shown in SEQ ID NO: 13; The genome DNA sequence of the fertility gene AetFRG1 in grass is shown in SEQ ID NO:14, the CDS sequence is shown in SEQ ID NO:15, and its protein sequence is shown in SEQ ID NO:16; the fertility gene SbFRG1 in sorghum The genomic DNA sequence is as shown in SEQ ID NO:17, the CDS sequence is as shown in SEQ ID NO:18, and its protein sequenc...

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Abstract

The invention discloses a wheat fertility recovery gene and an application thereof, belongs to the field of plant biotechnology, and in particular relates to a recessive genic male sterility gene and cloning of a promoter of the gene as well as an application of the gene in crossbreeding. Through flow cytometry and a high-throughput sequencing method, the fertility recovery gene FRG1 is successfully cloned; and with the application of the gene, male fertility of a Lanzhou genic male sterile mutant or an allelic mutant thereof can be completely recovered, so that a foundation is laid for the construction of a novel wheat crossbreeding technique system. Meanwhile, more possibilities are offered for solving such technical bottleneck problems in existing 'three-line' and 'two-line' wheat crossbreeding technologies which are unstable in sterile line fertility, limited in resource of hybrid varieties, complex in seed production technique, high in seed production cost and the like. The gene and sterile line propagation and maintaining method provided by the invention has great significance and application value of wheat crossbreeding work.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to the cloning of a plant recessive nuclear male sterility gene, its male sterile line propagation method and its application in hybrid breeding, and more specifically relates to a wheat recessive male sterile gene Cloning of genes and their promoters, and their application in crossbreeding. technical background [0002] Heterosis is a common phenomenon in the biological world. Hybrid breeding is the main way to select and breed new varieties, and is the most important method in modern breeding. Compared with rice, corn, sorghum, etc., the research on the use of heterosis in wheat is relatively lagging behind, and the growth of its average yield has also tended to stagnate in the past ten years, or even declined for many years. Wheat is a self-pollinated crop. The core issue of wheat heterosis utilization is the technical system for efficient production of wheat hybri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00
CPCC07K14/415C12N15/8287C12N15/8289C12N15/52C12N15/8231C12N15/8209C12N2830/007C12N2830/008C12N2830/65
Inventor 马力耕王峥李健何航陈少霞邓兴旺
Owner BEIJING NEXT GENERATION HYBRID WHEAT BIOTECHNOLOGY CO LTD
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