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Artificial fixed-point rice dense and erect panicle (DEP1) gene mutant body and application thereof

A technique of erecting dense ears and site-directed mutation, applied in genetic engineering, application, climate change adaptation, etc., can solve the problem of uncertainty of high yield effect, maximum potential, etc.

Inactive Publication Date: 2017-02-22
云南纳博生物科技有限公司
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AI Technical Summary

Problems solved by technology

[0004] The allele of DEP1 high-yield type widely used in production is consistent with the genotype of Shennong 265. However, this high-yield genotype is produced under natural mutation conditions. Whether the high-yield effect reaches the maximum potential of the DEP1 gene is still unclear. Sure

Method used

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  • Artificial fixed-point rice dense and erect panicle (DEP1) gene mutant body and application thereof
  • Artificial fixed-point rice dense and erect panicle (DEP1) gene mutant body and application thereof
  • Artificial fixed-point rice dense and erect panicle (DEP1) gene mutant body and application thereof

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Experimental program
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Effect test

Embodiment 1

[0030] The base sequence of the DEP1 gene is shown in SEQ ID NO.8.

[0031] The base sequence of exon 5 (wild type) of DEP1 gene is shown in SEQ ID NO.9, see Figure 8 .

[0032] In this example, based on CRISPR / Cas9 technology, the target of exon 5 of DEP1 gene was designed, and two targets AGCTGCGGTTGCAACGGCTG with close and consistent sequences were selected, and the first target was located at base 499-519 of exon 5 base, and the second target is located at the 530-549th base of exon 5. In this example, the target sequence is synthesized,

[0033] Oligo15'- GGCA AGCTGCGGTTGCAACGGCTG-3',

[0034] Ologo2: 5'-AAACAGCTGCGGTTGCAACGGCTG-3'.

[0035] In this example, the binary vector Pcambia1300 containing the cas9 expression cassette (35S promoter) and the guid RNA expression cassette (rice OsU3 promoter) was used as the backbone vector for vector construction, and the target sequence was inserted into the backbone vector. The specific method is as follows: Anneal and ph...

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Abstract

The invention discloses an artificial fixed-point rice dense and erect panicle (DEP1) gene mutant body and application thereof. The rice DEP1 gene mutant body as well as a corresponding allele is obtained by modifying a 5th exon of a DEP1 gene by adopting a CRISPR / Cas9 gene targeting modification technology, so that a base of the 5th exon is replaced, lost and inserted. The artificial fixed-point rice DEP1 gene mutant body and the application thereof disclosed by the invention have the benefits that the artificial mutation is performed on the 5th exon of the rice DEP1 gene by adopting the CRISPR / Cas9, and the mutant body capable of obviously improving the rice yield and the allele are screened; the DEP1 gene mutant body disclosed by the invention as well as the corresponding allele can improve the yield of rice plants by 13-51 percent and is even superior to a natural mutation type.

Description

technical field [0001] The invention relates to the field of rice genome editing and breeding, in particular to an artificial site-directed mutant of rice erect dense panicle DEP1 and its application. Background technique [0002] Rice is one of the important food crops in my country, and rice production accounts for about 40% of food crops. Increasing rice production is of great significance to ensuring national food security. There are two main periods for the improvement of rice yield in China: the first period is the 1950s, when dwarf breeding, that is, the "dwarf revolution" or "green revolution", and the second period is the hybridization since the 1970s. Widespread promotion of rice. Chen Wenfu, a rice expert, believes that ultra-high-yield breeding combining the use of heterosis and ideal plant type will surely be a breakthrough point for increasing rice yield in the future. [0003] Erect panicle is one of the ideal plant types for high-yield rice varieties. The ...

Claims

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Application Information

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IPC IPC(8): C12N15/29A01H5/00
CPCC07K14/415C12N15/8261Y02A40/10
Inventor 张业胜张如董扬
Owner 云南纳博生物科技有限公司
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