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Method for reducing plant height of tetraploid wild rice by inhibiting expression of SD1 gene

A tetraploid wild rice and gene technology, applied in biochemical equipment and methods, genetic engineering, plant genetic improvement, etc., can solve problems such as unfavorable field dense planting and harvesting

Active Publication Date: 2021-05-18
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the tetraploid wild rice Oryza alta has the advantages of large biomass, strong adaptability, and strong stress resistance. However, because it has not been artificially domesticated, the plant height is 2.7m, which is not conducive to dense planting and harvesting in the field.

Method used

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  • Method for reducing plant height of tetraploid wild rice by inhibiting expression of SD1 gene
  • Method for reducing plant height of tetraploid wild rice by inhibiting expression of SD1 gene
  • Method for reducing plant height of tetraploid wild rice by inhibiting expression of SD1 gene

Examples

Experimental program
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Effect test

Embodiment 1

[0057] Example 1, Selection of Tetraploid OaSD1 Target Site and Construction of CRISPR / Cas9 Genome Editing Vector

[0058] 1. Selection of target sequence

[0059] The 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 SD1 in Oryza alta, the gene located on chromosome 1 of the CC subgenome is named OaSD1-CC (nucleotide sequence shown in SEQ ID No.1), and the other gene located on The gene on chromosome 1 of the DD subgenome is named OaSD1-DD (the nucleotide sequence is shown in SEQ ID No.2).

[0060] OaSD1-CC consists of 3131 nucleotides, and its nucleotide sequence is shown in SEQ ID No.1. The reading frame is the 190th-3106th nucleotide of SEQ ID No.1, consisting of 3 exons, wherein the 190th-702nd of SEQ ID No.1 is the first exon, and the 812nd The position-1135th is the second exon, the 2797th-3106th is the third exon, and the rest are intron sequences. Its C...

Embodiment 2

[0093] Example 2, Creation and phenotypic determination of tetraploid wild rice O.alta plants with SD1 gene knockout

[0094] 1. Cultivation of tetraploid wild rice O.alta plants with SD1 gene knockout

[0095] The plasmid CRISPR-OaSD1-2Target 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 OaSD1-CC and OaSD1-DD were amplified with specific primers.

[0096] The primer pair used to amplify OaSD1-CC consists of OaSD1-CC-F and OaSD1-CC-R:

[0097] OaSD1-CC-F:5'-CAGCACCAACCAACATCAGC-3';

[0098] OaSD1-CC-R: 5'-TGAGCTGCTGTCCGCGAAGA-3'.

[0099] The primer pair used to amplify OaSD1-DD consists of OaSD1-DD-F and OaSD1-DD-R:

[0100] OaSD1-DD-F:5'-GCCATCATCTCATCCGACTC-3';

[0101] OaSD1-DD-R: 5'-GGAGCTGCTGTCCGCGAAGA-3'.

[0102] The elec...

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Abstract

The invention discloses a method for reducing the plant height of tetraploid wild rice by inhibiting expression of an SD1 gene. The specific embodiment of the invention proves that by editing a green revolution gene SD1, rapid domestication of the plant height can be realized, the plant height of tetraploid wild rice Oryza alta can be reduced, the tetraploid wild rice Oryza alta is convenient to harvest, besides, the lodging-resistant capability is strong, and the tetraploid wild rice Oryza alta is suitable for close planting.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for reducing the plant height of tetraploid wild rice by inhibiting the expression of SD1 gene. Background technique [0002] After a long period of domestication, there are only two species of cultivated rice currently used for production: Oryza sativa in Asia and Oryza glaberrima in Africa, while there are as many as 25 species of wild rice. Among them, the tetraploid wild rice Oryza alta has the advantages of large biomass, strong adaptability, and strong stress resistance. However, because it has not been artificially domesticated, the plant height is 2.7m, which is not conducive to dense planting and harvesting in the field. It is of great significance to reduce plant height to cultivate polyploid rice crops with strong lodging resistance and suitable for dense planting and management in the field. Contents of the invention [0003] The technical problem ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/29C12N15/82C07K14/415A01H5/00A01H6/46
CPCC12N15/113C12N15/8218C12N15/8261C07K14/415
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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