Application of transcription factor gene OsBEAR1 of oryza sativa to elongation of bred coleoptiles or varieties of oryza sativa suitable for direct sowing in fields

A technology of rice transcription factors and coleoptiles, applied in the fields of application, genetic engineering, plant products, etc., can solve the problems of limited application, unfavorable rice germination, hypoxia, etc., and achieve the effect of improving germination and emergence rate

Active Publication Date: 2020-06-19
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Flooding is the most important limiting factor for direct seeding of rice. Flooding will cause hypoxia, and cells will produce and accumulate toxic substances during hypoxia, which is not conducive to the germination of rice.
The seedling rate of many rice varieties will be severely reduced under flooding conditions, which limits their application in direct seeding

Method used

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  • Application of transcription factor gene OsBEAR1 of oryza sativa to elongation of bred coleoptiles or varieties of oryza sativa suitable for direct sowing in fields
  • Application of transcription factor gene OsBEAR1 of oryza sativa to elongation of bred coleoptiles or varieties of oryza sativa suitable for direct sowing in fields
  • Application of transcription factor gene OsBEAR1 of oryza sativa to elongation of bred coleoptiles or varieties of oryza sativa suitable for direct sowing in fields

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: Rice OsBEAR1 Knockout

[0029] one, OsBEAR1 Construction of knockout vector

[0030] Using the online tool (http: / / skl.scau.edu.cn / ) from OsBEAR1 Select the target sequence to be knocked out from the specific segment of the gene sequence: "5'-GGCTCTCGTCGACCGTATCA-3'" (SEQ ID NO: 4).

[0031] Select the sgRNA expression vector "pYLsgRNA-OsU6a" with the promoter OsU6a, and synthesize the linker primer corresponding to the target sequence:

[0032] Upstream primer: 5′-GCCGGGCTCTCGTCGACCGTATCA-3′

[0033] Downstream primer: 5′-AAACTGATACGGTCGACGAGAGCC -3′

[0034] The two primers are annealed to form small DNA fragments.

[0035] Use primers UF: 5′-CTCCGTTTTACCTGTGGAATCG -3′ and gR-R: 5′-CGGAGGAAAATTCCATCCAC-3′ to perform inverse PCR with the pYLsgRNA-OsU6a vector as a template to linearize it. The linearized pYLsgRNA-OsU6a was digested with the restriction endonuclease Bsa I, then the annealed DNA fragment of the target sequence with the linker was add...

Embodiment 2

[0059] Example 2: OsBEAR1 Coleoptile Growth Experiment of Knockout Plants

[0060] The flow process of the growth experiment used in the present invention is: with acceptor rice (WT) and OsBEAR1 The CRISPR-knockout rice (CRI-BEAR) was used as the experimental material. The plump and good seeds were first baked at 42°C for 24 hours, then soaked in distilled water at 37°C for 12 hours, and then the seeds were germinated in a semi-exposed shallow water layer for 2 days. Transfer the germinated seeds to 24 degrees, and cultivate them with distilled water in a photoperiod culture environment with 13 hours of light and 11 hours of darkness, observe and take pictures on the third day of photoperiod culture, randomly select 8 plants from each material, and record the coleoptile phenotype ( Figure 4 ), the corresponding coleoptile length statistics such as Figure 5 , the average length of coleoptiles of recipient rice (WT) was 0.61cm, and the average length of coleoptiles of knock...

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Abstract

The invention discloses a gene OsBEAR1 for adjusting and controlling growth and development of coleoptiles of oryza sativa, and an application of the gene OsBEAR1. The MSU ID of the gene is ''LOC_Os11g15210'', and the gene is used for coding an amino acid sequence as shown in SEQ ID NO:3. By a gene editing method, the gene mutant is knocked out in Oryza sativa L. cv. Nipponbare, growth and development of the coleoptiles of oryza sativa seedlings after OsBEAR1 knockout are accelerated, and the length of the coleoptiles of the oryza sativa seedlings after OsBEAR1 knockout is notably longer thanthat of coleoptiles of receptor oryza sativa seedlings. The elongation of the coleoptiles facilitates sprouting of oryza sativa and direct sowing in the fields. According to the gene OsBEAR1 disclosedby the invention, a base is provided for reformation and breeding of the varieties of oryza sativa suitable for direct sowing in the fields.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to rice coleoptile growth and development genes OsBEAR1 and its application. Background technique [0002] Rice is an important food crop for human beings. People have been pursuing simple, economical and efficient production methods. Rice direct seeding technology is an important part of this high-efficiency cultivation. Flooding is the most important limiting factor for direct seeding of rice. Flooding will cause hypoxia, and cells will produce and accumulate toxic substances during hypoxia, which is not conducive to the germination of rice. The seedling rate of many rice varieties will be severely reduced under flooding conditions, which limits their application in direct seeding. [0003] The coleoptile is the plant's protective tissue that protects the germ from damage when it emerges from the soil. At the same time, rice coleoptiles can grow in a hypoxic environment, which plays a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8218C12N15/8261
Inventor 陈涛滕炎桐
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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