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Application of substance inhibiting expression of OaGS3 gene to regulating and controlling grain length of tetraploid wild rice

A tetraploid wild rice, gene expression technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of increasing the risk of stable grain yield, slow increase in crop yield, and sharp reduction of cultivated land area.

Active Publication Date: 2021-04-20
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the intensification of urbanization has led to a sharp decrease in arable land, frequent extreme weather has increased the risk of stable grain production, and the increase in crop yield brought about by traditional breeding techniques has been slow

Method used

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  • Application of substance inhibiting expression of OaGS3 gene to regulating and controlling grain length of tetraploid wild rice
  • Application of substance inhibiting expression of OaGS3 gene to regulating and controlling grain length of tetraploid wild rice
  • Application of substance inhibiting expression of OaGS3 gene to regulating and controlling grain length of tetraploid wild rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1, Selection of Tetraploid OaGS3 Target Site and Construction of CRISPR / Cas9 Gene Editing Vector

[0056] 1. Selection of target sequence

[0057] The tetraploid tall-stalked tetraploid wild rice Oryza alta is an allotetraploid, and most genes have copies in the CC and DD subgenomes. Through comparative analysis, there are two homologous genes of OaGS3 in Oryza alta, the gene located on chromosome 3 of the CC subgenome is named OaGS3-CC, and the gene located on chromosome 3 of the DD subgenome is named OaGS3-DD (The nucleotide sequence is shown in sequence 2).

[0058] OaGS3-CC consists of 5926 nucleotides, and its nucleotide sequence is shown in SEQ ID NO:1. Its CDS sequence is shown in sequence 3, and the encoded protein sequence is shown in sequence 5.

[0059] OaGS3-DD consists of 6193 nucleotides, and its nucleotide sequence is shown in SEQ ID NO:2. Its CDS sequence is shown in sequence 4, and the encoded protein sequence is shown in sequence 6.

[006...

Embodiment 2

[0070] Embodiment 2, transformation tetraploid rice callus and phenotypic identification

[0071] 1. Breeding OaGS3 gene knockout tetraploid wild rice O.alta plants

[0072] The plasmid VK005-OaGS3gRNA obtained in Example 1 was transformed into Agrobacterium EHA105 by electric shock method, and the callus of tetraploid wild rice O. alta was infected with Agrobacterium, and the above callus was cultivated to obtain transformed plantlets. Genomic DNA of transformed shoots was extracted, and OaGS3-CC and OaGS3-DD were amplified with specific primers.

[0073] The primer pair used to amplify OaGS3-CC consists of OaGS3-CC-F and OaGS3-CC-R:

[0074] OaGS3-CC-F:5'-CCTCCGCCATTTATAATCCA-3';

[0075] OaGS3-CC-R: 5'-TATGCATTCGTGGTTTCAGC-3'.

[0076] The primer pair used to amplify OaGS3-DD consists of OaGS3-DD-F and OaGS3-DD-R:

[0077] OaGS3-DD-F:5'-GCTGCCTTTCCATCATCATT-3';

[0078] OaGS3-DD-R: 5'-ATGTTGGGCCATGCATATTT-3'.

[0079] The electrophoresis of the amplified product is sh...

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Abstract

The invention provides an application of a substance regulating and controlling an OaGS3 gene to regulating and controlling the yield / grain length of tetraploid wild rice. The specific embodiment of the invention proves that the grain length can be increased by editing the gene OaGS3, so that the tetraploid wild rice with higher yield is bred, and domestication and utilization of the tetraploid wild rice are promoted.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of a substance inhibiting OaGS3 gene expression in regulating the grain length of tetraploid wild rice. Background technique [0002] Rice is one of the most important food crops in the world. High and stable rice yields are of great significance to ensuring world food security. However, the intensification of urbanization has led to a sharp decrease in arable land, frequent extreme weather has increased the risk of stable grain production, and the increase in crop yield brought about by traditional breeding techniques has been slow. The issue of food security has become increasingly prominent and has gradually become a new challenge for people. [0003] The tall tetraploid wild rice O.alta is a perennial allotetraploid rice, mainly distributed in South America. Compared with diploid cultivated rice, tetraploid high-stalk tetraploid wild rice has many a...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N15/84C12N15/55A01H5/10A01H6/46
Inventor 李家洋余泓孟祥兵张静昆刘贵富荆彦辉陈明江
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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