Method for preparing photosensitive male-sterility material of rice and related gene

A male sterility and photosensitive technology, which can be used in botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., and can solve problems that have not yet been discovered

Active Publication Date: 2021-11-12
HUNAN HYBRID RICE RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in addition to the application of these genes in production, there are still many undiscovered genes that control photothermo-sensitive male sterility. If these genes can be deeply excavated and their functions studied, it will be more comprehensively expanded and deepened. The understanding and understanding of the mechanism of photothermosensitive male sterility has important guiding significance and application value for the selection and creation of new and excellent stable two-line sterile lines

Method used

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  • Method for preparing photosensitive male-sterility material of rice and related gene
  • Method for preparing photosensitive male-sterility material of rice and related gene
  • Method for preparing photosensitive male-sterility material of rice and related gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1, Selection of Rice RMS1 Gene Target Site and Construction of Knockout Vector

[0102] The inventors of the present invention discovered a gene related to photosensitive male sterility—the RMS1 (Reverse Male Sterility) gene from rice Wuyunjing 7. Wherein, the coding sequence of the RMS1 gene is shown in SEQ ID No.2 in the sequence listing, encoding a protein RMS1 consisting of 345 amino acid residues, and its amino acid sequence is shown in SEQ ID No.1 in the sequence listing. The gDNA of the RMS1 gene has a full length of 2623bp, contains 3 exons and 4 introns, and its nucleotide sequence is shown in SEQ ID No.3 in the sequence listing.

[0103] In this example, the rice RMS1 gene was knocked out by CRISPR / Cas9 gene editing technology to obtain mutant 9522 with a photosensitive male sterile phenotype 38740-4 、9522 38740-5 and 9522 38740-11 , mutant 9522 38740-4 、9522 38740-5 and 9522 38740-11 All RMS1 gene knockout rice. The specific operation method is...

Embodiment 2

[0123] Example 2 The acquisition of RMS1 mutant rice material and its phenotype analysis

[0124] 1. Obtaining RMS1 mutant rice materials

[0125] Using the method of Agrobacterium-mediated transformation of rice callus, the RMS1 gene site-directed editing vector pYLCRISPR / Cas9-MT-RMS1 was used to transform the callus of rice variety Wuyunjing 7 (formerly coded as 9522, hereinafter referred to as 9522), and screened. Positive mutants were identified.

[0126] 2. Detection of fixed-point editing

[0127] Homozygous mutants of the three types of mutations in which the RMS1 gene was knocked out were obtained by PCR detection, and they were named homozygous mutants 9522 38740-4 , homozygous mutant 9522 38740-5 , homozygous mutant 9522 38740-11 . The sequencing results showed that (such as Figure 6 Show):

[0128] RMS1 gene (wild type) in homozygous mutant 9522 38740-4 There was a mutation in the RMS1 gene CDS 128-129 missing 2 bases, which resulted in a shift of the RMS1 ...

Embodiment 3

[0139] Example 3 Analysis of photosensitivity characteristics of RMS1 mutant rice materials

[0140] 1. Analysis of photosensitivity characteristics of RMS1 mutant rice materials

[0141] RMS1 mutant material homozygous mutant 9522 planted under natural conditions in Lingshui, Hainan (18°51′23″N, 110°5′6″E) 38740-4 T 2 generation, homozygous mutant 9522 38740-5 T 2 generation and wild-type rice 9522, it was found that the homozygous mutant 9522 38740-4 T 2 generation, homozygous mutant 9522 38740-5 T 2 The seed setting rate of generation plants was low, 4.56% and 3.13%, respectively, while the seed setting rate of wild-type rice 9522 was 95.6% (201812-201904); ) under natural conditions to grow the homozygous mutant 9522 38740-4 T 3 generation, homozygous mutant 9522 38740-5 T 3 generation and wild-type rice 9522, it was found that the homozygous mutant 9522 38740-4 T 3 generation, homozygous mutant 9522 38740-5 T 3 The seed setting rates of the first generation...

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Abstract

The invention discloses a method for preparing a photosensitive male-sterility material of rice and a related gene. The method for preparing photosensitive male-sterility rice comprises the following step: reducing the abundance of a protein RMS1 in target rice, reducing the activity of the protein RMS1 in the target rice or reducing the content of the protein RMS1 in the target rice to obtain the photosensitive male-sterility rice. The protein RMS1 is A1) or A2) as follows: A1) an amino acid sequence of the protein RMS1 is shown as SEQ ID No. 1 in a sequence listing; and A2) a homologous protein which has 98% or more of identity with the A1) and is derived from rice. According to the method, the photosensitive male-sterility material of the rice is obtained by controlling a RMS1 gene of the rice and a coding protein of the RMS1 gene, and conversion of the rice fertility under different illumination conditions is achieved.

Description

technical field [0001] The invention relates to the field of biotechnology breeding, in particular to a method for preparing rice photosensitive male sterile materials and related genes. Background technique [0002] Hybrid breeding technology has played a pivotal role in increasing rice production in my country and the world, and is an important guarantee for my country's food security and sustainable agricultural development. The two-line male sterile lines can be combined freely and the seed production procedure is simplified, but the fertility is greatly affected by the external environment, and there are risks in seed production. The selection of light-temperature-sensitive male sterile lines is the development and application of the two-line hybrid rice breeding technology system At present, temperature-sensitive male sterile lines are widely used in production, which are mainly controlled by temperature. In a relatively variable temperature environment, the length of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C07K14/415C12N15/29C12N15/113C12N9/22A01H5/00A01H6/46
CPCC12N15/8289C12N15/8213C12N15/8261C07K14/415A01H5/00A01H6/46C12N9/22C12N15/113C12N15/82
Inventor 李莉邱牡丹李懿星张大兵宋书锋王天抗
Owner HUNAN HYBRID RICE RES CENT
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