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A kind of rice fertility gene saw1 and its application

A fertility gene and rice technology, applied in the field of genetic engineering, can solve the problems of high cost of seed production, unstable photo-temperature-sensitive sterility, lack of germplasm resources, etc., and achieve the effect of stable fertility

Active Publication Date: 2021-06-15
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the current technical bottlenecks such as the lack of germplasm resources of cytoplasmic sterile lines, the instability of photothermo-sensitive sterility, and the high cost of seed production, scientists are trying to use a new hybrid breeding technology, which will make full use of recessive cytoplasmic sterile lines. Fertility genes, construct a stable sterile line whose fertility is not affected by the environment, remove the constraints of environmental factors on hybrid breeding, and eliminate potential risks in production

Method used

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  • A kind of rice fertility gene saw1 and its application
  • A kind of rice fertility gene saw1 and its application
  • A kind of rice fertility gene saw1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, mutagenesis screening rice male sterile mutant (saw1)

[0047] Indica rice variety Huanghuazhan (HHZ) passed Co 60 γ-ray radiation mutagenesis, screening mutant saw1.

[0048] The reciprocal cross experiment between mutant saw1 and wild type HHZ confirmed that mutant saw1 is a male sterile mutant.

[0049] Take the soon-to-flower florets of the mutant saw1, squeeze the anthers with tweezers, and add 1% I 2 -KI staining found that the pollen was corrupted. However, the plant height, tiller number, heading date and leaf number of the mutant saw1 were the same as those of the wild-type HHZ (the plant type at the heading stage was as follows: figure 1 , the anther shape is as figure 2 , the results of pollen staining analysis are as follows image 3 ).

[0050] In addition, the phenotype of mutant saw1 was stable and not affected by light and temperature after planting in early season (long sunshine + high temperature) and late season (short sunshine + ...

Embodiment 2

[0051] Embodiment 2, the acquisition of rice fertility-related genes

[0052] 1. InDel marker linkage analysis

[0053] F 1 , and F after selfing 2 Isolated populations (200 strains). in F 2 From the population, 10 mutants whose phenotype was extremely similar to mutant saw1 were selected, and their leaves were used to extract DNA.

[0054] The method of extracting single-plant DNA refers to the method of SDS micro-extraction of rice DNA (Zhou et al., 2016, Current Protocols in Plant Biology 1, 29-42).

[0055] Linkage analysis was performed using InDel markers covering the rice genome, and it was initially found that the genes controlling rice fertility existed between markers 26621 and 28076. Then design the InDel mark between 26621 and 28076 by yourself, and use F 3 The large population was further fine-tuned, and the mapping interval was narrowed, and the target gene was finally narrowed between markers 26785 and 26819. According to the published genome sequence of ...

Embodiment 3

[0066] Example 3, Construction of Knockout Vector of SAW1 Gene and Obtaining and Identification of Transgenic Plants

[0067] (1) Referring to the sequence of the indica rice HHZ genome, select 3 targets in the genomic region corresponding to the SAW1 exon ( Figure 6 ), these three targets have common features, 3' end has NGG (N is any base of A, T, C, G). Primer pairs were designed for these three targets: Primer1 (SEQ ID NO.24) / Primer2 (SEQ ID NO.25), Primer3 (SEQ ID NO.26) / Primer4 (SEQ ID NO.27), Primer5 (SEQ ID NO.27) NO.28) / Primer6 (SEQ ID NO.29).

[0068] The CRISPR / Cas9 vector pYLCRISPR / Cas9Pubi-SAW1 containing the above three targets was constructed according to the method of pYLCRISPR / Cas9Pubi-H vector construction (Ma et al., 2015, Molecular Plant8, 1274-1284). The recombinant plasmid pYLCRISPR / Cas9Pubi-SAW1 was transformed into Agrobacterium EHA105 strain by electric shock method to obtain the recombinant strain, and the plasmid was extracted for PCR and enzyme d...

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Abstract

The invention discloses a rice fertility gene SAW1 and its application. The nucleotide sequence of the gene SAW1 is shown in SEQ ID NO.1, and the encoded amino acid sequence is shown in SEQ ID NO.2. Knocking out the gene SAW1 can abort rice anthers, and can be applied to the creation of sterile lines and the preparation of hybrid seeds. The male sterile line produced based on the mutation of gene SAW1 has stable fertility and is not affected by environmental conditions. It provides necessary elements for the construction of a new hybrid breeding system and has good application value and prospects.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering. More specifically, it relates to a rice fertility gene SAW1 and its application. Background technique [0002] Rice is one of the most important food crops in the world, providing about 21% of the energy intake needs of the world's population. my country is not only a big rice producer in the world but also a big consumer. The import volume of rice in our country is increasing continuously. In 2015 alone, the annual import volume exceeded 3 million tons for the first time. At present, we are facing the severe challenge of how to increase the total rice output and ensure food security under the condition that the rice planting area remains basically unchanged or decreases year by year. Hybrid breeding is an effective way to increase crop yield. Hybrids usually have obvious advantages over common varieties in terms of yield, stress resistance, and quality. The breeding cycle of hybrid...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N1/21C12N15/11A01H5/00A01H6/46C12R1/01
CPCC07K14/415C12N15/8218C12N15/8289
Inventor 祝钦泷刘耀光陈乐天方瑞秋王斌陈法铭韩靖鸾
Owner SOUTH CHINA AGRI UNIV
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