Application of OsGA3ox1 gene to rice male sterile strain line creation

A technology of male sterility and rice, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of incomplete fertility, unseen, high cost, etc.

Active Publication Date: 2017-10-03
江西省超级水稻研究发展中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are still problems such as incomplete fertility, high cost, or serious deterioration of other agronomic traits in the prepara...

Method used

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  • Application of OsGA3ox1 gene to rice male sterile strain line creation
  • Application of OsGA3ox1 gene to rice male sterile strain line creation
  • Application of OsGA3ox1 gene to rice male sterile strain line creation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1. A method for creating male sterile rice lines by knocking out the gibberellin synthetic gene OsGA3ox1

[0068] The knockout of the OsGA3ox1 gene and the acquisition of male sterile plants are accomplished through the following steps:

[0069] 1) Design sgRNA guide sequence

[0070] Using the online tool CRISPR-P (http: / / cbi.hzau.edu.cn / crispr / ), according to the method provided by the developer, design the sg RNA guide sequence for the OsGA3ox1 gene, and from the results provided by the software, according to the sequence specificity and the position of the sequence to select a suitable guide sequence, the sgRNA guide sequence selected in this embodiment is the sequence shown in SEQ ID No.3 and SEQ ID No.4.

[0071] 2) Synthesize oligonucleotides for cloning according to the guide sequence and vector requirements

[0072] This example uses the CRISPR / Cas9 vectors pCUbi1390Cas9-U3 and pCUbi1390Cas9-U6 based on the vector pCAMBIA1390 (constructed by...

Embodiment 2

[0123] Example 2, Fertility Recovery of OsGA3ox1 Gene Knockout Mutants

[0124] At the heading stage of rice, GA containing gibberellin substances can be used at 8 o'clock in the morning 3 The solution can be sprayed on the panicle of rice to restore fertility, the concentration can be 45mg / L gibberellin, spray 4 times, every 667m 2 Within 18g of the dosage, the gibberellin solution should contain a certain amount of surfactant such as 0.01% Tween-20 to enhance the adhesion of the agent on the leaf surface. When using kasalath as the background, use GA with a concentration of about 30mg / L 3 Spray when the ears are just coming out.

Embodiment 3

[0125] Example 3, Fertility Restoration of OsGA3ox1 Gene Knockout Mutants

[0126] At the heading stage of rice, GA containing gibberellins can be used at 5 p.m. 3 The solution can be sprayed on the ear of rice to restore fertility, the concentration can be 60mg / L gibberellin, spray once, every 667m 2 Within 30 g of the dosage, the gibberellin solution should contain a certain amount of surfactant such as 0.03% Tween-20, so as to enhance the adhesion of the medicament on the leaf surface.

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Abstract

The invention belongs to the technical field of rice breeding, and relates to a method for creating a male sterile strain line by using a gibberellin synthesis gene. The sterile line creation belongs to the key for the rice hybrid production. In order to overcome the defects of the existing three-line and two-line sterile line, the invention provides a novel sterile line and a creation method thereof, and concretely discloses a method of creating the rice male sterile strain line by using the rice gibberellin synthesis gene OsGA3ox1 and a fertility restoration method of the sterile line. The inventio provides fire-new application of the gibberellin synthesis gene OsGA3ox1. The method for realizing self breeding through external gibberellin spraying is provided for the hybrid production; the fertility is not influenced by the environment conditions; the novel dual-purpose system overcomes the defect of high risk of the existing photo-thermo sensitive dual-purpose sterile line due to the influence of extreme weather.

Description

technical field [0001] The invention belongs to the technical field of rice breeding, and relates to a brand-new application of rice gibberellin synthetic gene OsGA3ox1 and a method for creating a related novel rice sterile line, in particular to the use of OsGA3ox1 gene in the creation of rice male sterile lines The application of OsGA3ox1 gene, the method for creating rice male sterile lines and the method for restoring the fertility of rice male sterile lines created by this method. Background technique [0002] Obtaining sterile lines is the basis of heterosis utilization in rice. At present, the male sterile lines used in rice production are mainly cytoplasmic (cytoplasmic-nuclear interaction) male sterile lines and photothermosensitive genic sterile lines. Related seed production is also correspondingly divided into "three-line" and "two-line" seed production. The "three-line" system has the problem of unfree combination due to the limitation of the restoration relat...

Claims

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Application Information

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IPC IPC(8): C12N9/02A01G7/06A01H1/02A01H5/00
CPCA01G7/06A01H1/02C12N9/0071C12N15/8213C12N15/8289C12Y114/11015
Inventor 龙起樟万建林黄永兰王会民唐秀英芦明
Owner 江西省超级水稻研究发展中心
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