Related protein at early blossoming stage of rice as well as encoding gene thereof

A technology related to protein and rice, applied in genetic engineering, plant genetic improvement, angiosperms/flowering plants, etc., can solve the problems of high production cost and low seed production yield, and achieve low cost, high breeding efficiency, identification and selection simple effect

Active Publication Date: 2018-05-08
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the problems of low seed production yield and high production cost caused by the lack of flowering time between the sterile line and the restorer line existing in the current rice three-line seed production, the purpose of the present invention is to provide a rice early flowering time-related protein

Method used

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  • Related protein at early blossoming stage of rice as well as encoding gene thereof
  • Related protein at early blossoming stage of rice as well as encoding gene thereof
  • Related protein at early blossoming stage of rice as well as encoding gene thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Early flowering gene screening in the early flowering mutant emf1 of the present invention

[0048] (1) Test materials

[0049] (1) Emf1, an early flowering mutant, is an early flowering mutant that the inventors screened from an EMS (ethyl methylsulfonate) mutagenesis library constructed on the background of the indica rice maintainer Yixiang 1B. The mutant was backcrossed with Yixiang 1B for multiple generations, and the trait of early flowering was genetically stable.

[0050] (2) Early flowering mutant emf1 and wild type Yixiang 1B (see figure 1 and figure 2 ), and 02428 (japonica rice) were obtained from the Genetic Research Laboratory of the Rice Research Institute of Sichuan Agricultural University.

[0051] (2) Test method

[0052] (1) Hybridization of emf1 and 02428 to construct F 1 Generation population, and selfing to get F 2 generation populations for genetic mapping. Backcross emf1 with Yixiang 1B to construct BC 1 f 3 Generation populat...

Embodiment 2

[0066] Example 2 The gene knockout verification test of the early flowering time selection gene OsEFT1 of the present invention

[0067] (1) Test materials

[0068] The Escherichia coli competent DH5α used in this experiment was purchased from Beijing Quanshijin Biological Co., Ltd., and the Agrobacterium EHA105 strain was donated by Wang Wenming's research group at the Rice Research Institute of Sichuan Agricultural University.

[0069] (2) Test method

[0070] 1. Construction of CRISPR / Cas9-OsEFT1 gene knockout vector

[0071] Using F: 5'-ATGCGTCGCGTCACGGTG-3' and R: 5'-CAGGAGCAGCCGCCGGCC-3' as primers, the cDNA of mutant emf1, Yixiang 1B, and Zhonghua 11 were used as templates to amplify the gene respectively, and found that the gene The highly homologous nucleotide sequences in the coding region of the indica rice Yixiang 1B and the japonica rice Zhonghua 11 indicated that the protein encoded by the gene had no difference between the indica rice Yixiang 1B and the japoni...

Embodiment 3

[0112] Example 3 Transcriptome sequencing test of early flowering mutant emf1 of the present invention

[0113] RNA-seq sequencing splicing was carried out at Shanghai Ouyi Biological Co., Ltd. The specific steps are as follows:

[0114] (1) RNA extraction and library construction

[0115] (1) On the previous day, select the plants whose spikelets had bloomed to 1 / 3 to be marked, and collect the mutant emf1, Wild-type Yixiang 1B spikelets were quick-frozen in liquid nitrogen and stripped on ice, and three biological replicates were set up. Use kit mirVana TM miRNA ISOlation Kit, Ambion-1561 extracts total RNA, the sample quality requirement is ≥5 μg, and the purity requirement is OD: 1.8-2.2.

[0116] (2) Strand-specific transcription building library

[0117]After extracting the total RNA of the pulp and digesting the DNA with DNase, the eukaryotic mRNA is enriched with Oligo(dT) magnetic beads; the mRNA is broken into short fragments with specific reagents, and the brok...

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Abstract

The invention discloses related protein at the early blossoming stage of rice, and belongs to the field of rice gene engineering. The protein consists of an amino acid sequence as shown in SEQ ID No.1. The invention also discloses the corresponding encoding gene consisting of a nucleotide sequence as shown in SEQ ID No.2. The invention also discloses a gene at the early blossoming stage of the rice. Firstly, the glume-opening time controlled by the early blossoming gene is 1.5 to 2 hours in advance than the glume-opening time of the corresponding wild type, the glume-opening time in-advance effect is remarkable, and a new gene and an approach are provided for increasing the yield of rice seed production; secondly, the character at the early blossoming stage is controlled by a recessive gene, and identification and selection of transgenic offspring or hybridization breeding offspring are simple; and the mutant at the early blossoming stage provides a good research material for researchand application of the gene at the early blossoming stage.

Description

technical field [0001] The invention belongs to the field of rice genetic engineering, and in particular relates to a rice early flowering time-related protein, a gene encoding the protein, and their use in rice three-line seed production. Background technique [0002] The application of hybrid rice has greatly increased the yield per unit area of ​​rice, but due to the low yield and high cost of hybrid rice seed production, the cost of purchasing seeds for farmers is correspondingly high, which increases the production cost. Increasing seed production per unit area is the key to reducing hybrid rice seed production costs. [0003] The main limiting factor for hybrid rice seed production is the low outcross seed setting rate of sterile lines, and the flowering habit of male sterile lines is one of the determinants of outcross seed setting rate (Yuan Longping. Hybrid Rice Science. China Agricultural Press, 2002, 246-279). Due to the negative effect of the sterile cytoplasm,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/113C12N15/82A01H5/02A01H6/46A01H1/02
CPCA01H1/02C07K14/415C12N15/113C12N15/8213C12N15/827C12N2310/10
Inventor 徐培洲吴挺开陶宇孙朝伟吴先军张红宇陈晓琼廖泳祥邓小书余世聪
Owner SICHUAN AGRI UNIV
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