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
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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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