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Application of rice OsFD2 gene in regulation and control of seed size

A technology for rice seeds and rice, which is applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problem that the molecular mechanism regulating the size of rice seeds has not been fully elucidated.

Active Publication Date: 2022-02-01
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, the molecular mechanism that regulates the size of rice seeds has not been fully elucidated, and there is fragmentation between the functions of each gene.

Method used

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  • Application of rice OsFD2 gene in regulation and control of seed size
  • Application of rice OsFD2 gene in regulation and control of seed size
  • Application of rice OsFD2 gene in regulation and control of seed size

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 Contains the construction and genetic transformation of target gene overexpression vector

[0038] 1. Design primers according to the sequence and cDNA sequence of the OsFD2 gene provided by NCBI, and clone the OsFD2 gene from the rice variety Zhonghua 11 by conventional methods. The sequence of the OsFD2 gene is shown in SEQ ID No.1, and the coding region of the OsFD2 gene The sequence is shown in SEQ ID No.2, and the encoded protein sequence is shown in SEQ ID No.3. The gene is located on chromosome 6, and its gene locus number in rice is LOC_Os06g50600 (Os06g0720900).

[0039] SEQ ID No. 1:

[0040]

[0041]

[0042]

[0043] SEQ ID No.2:

[0044]

[0045]SEQ ID No. 3:

[0046]

[0047]

[0048] 2. Select a suitable enzyme cutting site and digest the Pox vector. The cloned OsFD2 gene was connected to the Pox vector by homologous recombination. Subsequently, the ligation product was transformed into a competent DH5α and positive colo...

Embodiment 2

[0053] Construction and genetic transformation of embodiment 2 OsFD2 knockout vector

[0054] 1. Obtain the whole genome sequence (SEQ ID No.1) of OsFD2 through NCBI, through the target design in the website (CRISPR-GE(Genome Editing)-Liu YG Lab(http: / / skl.scau.edu.cn / ) Design and select targets with low off-target rate, and design corresponding target primers. The primer sequence is F: ggcaCCATCTGGTAGTACTCCTGG R: aaacCCAGGAGTACTACCAGATGG.

[0055] 2. Connect the target point to the U3 gRNA expression cassette to construct the CRISPR / gRNA vector ( figure 2 ).

[0056] 3. Clone the U3 gRNA expression cassette and connect it to the pYLCRISPR / Cas9-Mtmono binary vector ( image 3 ) to obtain the OsFD2 knockout vector.

[0057] 4. Using Agrobacterium tumefaciens-mediated genetic transformation to introduce the above-mentioned OsFD2 knockout vector into rice callus, and obtain plants fd2-1 and fd2-2 transformed from two different calli.

[0058] 5. The target position is amplif...

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Abstract

The invention discloses application of a rice OsFD2 gene in regulation and control of seed size, and belongs to the field of plant molecular biology. The invention provides application of OsFD2 protein or related biological materials thereof in regulation and control of rice seed size, and the OsFD2 protein is protein with an amino acid sequence of SEQ ID No.3. The related biological material is a nucleic acid molecule capable of expressing the OsFD2 protein, or an expression cassette, a recombinant vector, a recombinant bacterium or a transgenic cell line containing the nucleic acid molecule; and according to the invention, the function of the OsFD2 gene is deleted, so that the rice seeds are regulated and controlled to become larger, the OsFD2 gene is over-expressed, and the rice seeds are regulated and controlled to become smaller. Gene resources are provided for crop breeding, and the rice OsFD2 gene can be applied to rice breeding and has important application value.

Description

technical field [0001] The invention relates to the field of plant molecular biology, in particular to the application of a rice OsFD2 gene in regulating seed size. Background technique [0002] Rice is one of the most important food crops in the world, and its yield is crucial to human survival and development. Factors that determine rice yield include the number of grains per panicle, the number of effective tillers and grain weight. Among them, grain weight is a relatively stable genetic factor, which is not easily affected by the external environment, and is relatively easy to control in genetic breeding. Grain weight is mainly determined by grain size, controlled by polygenes, and is an important and complex agronomic trait. Rice grain size is mainly determined by glumes that limit grain growth. [0003] In recent years, research on rice seed size has been ongoing. Studies have shown that G-protein signaling pathway, plant hormones, transcription factors, ubiquitin-...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/84C12N15/55A01H5/10A01H6/46
CPCC07K14/415C12N15/8261C12N15/8205C12N15/8218C12N9/22
Inventor 姜大刚何荧庄楚雄
Owner SOUTH CHINA AGRI UNIV