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Rice salt-resistance-related gene onac103 and its encoded protein and its application

A salt stress and rice technology, applied in the fields of genetic engineering and molecular biology, can solve the problems of insufficient per capita arable land, inability to use saline-alkali land, and insufficient usable land area.

Active Publication Date: 2021-12-10
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although our country has a vast territory and a large land area, due to our large population, the per capita arable land area is insufficient, which makes our usable land area seriously insufficient.
At the same time, in arid and semi-arid areas, there are still large areas of unusable saline-alkali land

Method used

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  • Rice salt-resistance-related gene onac103 and its encoded protein and its application
  • Rice salt-resistance-related gene onac103 and its encoded protein and its application
  • Rice salt-resistance-related gene onac103 and its encoded protein and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Substrate of the Resistance of Salt Stress Related Genes ONAC103

[0020] In the present invention, the total RNA is extracted with TP309, and the total RNA is extracted with the reagent cartridge to reverse the RNA to cDNA. With cDNA as a template, the resolution of the amplification target can utilize the PRIMERSTAR HS DNAPOLYMERASE amplification of the target fragment in the present invention. The primer sequence is:

[0021] OE-ONAC103-FOR: GATGTTGGGTTCGGCCCC

[0022] OE-ONAC103-REV: TcagaatagctGaggagaagcccc

[0023] The response system of the PCR amplification is:

[0024]

[0025] The amplification conditions of PCR were 94 ° C for 3 min; 94 ° C denaturation 30s, 56 ° C, 10S, 72 ° C for 1 min, and the cycle was 35 cycles; 72 ° C extends for 5 min. 4 ° C insulation.

[0026] The PCR product was recovered by means of purification of glue recovery or PCR product, and the recovered PCR product was added to a tail. Easytaq DNA Polymerase is used when adding A....

Embodiment 2

[0030] Example 2: Agrobacterium transformation gains transgenic rice plants

[0031] Step 1: Rice healing organization ready

[0032] 1. Select the TP309 rice seed shell, placed in 50 ml of centrifuge tube, wash 3 times with sterile water, 1 min each time;

[0033] 2, use 75% ethanol to disinfect the seed three times, 1min each time, washed with sterile water;

[0034] 3, add 10% sodium hypochlorite solution, vacuum 8min, then placed on the flip mixer 30N / min, shake 20 min;

[0035] 4. Wash the sodium seed sodium seed surface with sodium seed surface, placed seeds on the sterilized filter paper;

[0036] 5. Use tweezers to inoculate seeds on NBD medium, and after 21 days under dark conditions, the bud treatment is carried out, and when the bud, pay attention to the germ sheath, gas roots, etc., only the wonderful callus in the growth state ( It is generally orange), and then cultivates 3-7D for Agrobacter.

[0037] Step 2: Agrobacterium dip

[0038] 1, a small amount of YEP medi...

Embodiment 3

[0060] Example 3: Analysis of transgenic plants overexpressing the ability to salt stress

[0061] figure 2 As one of the T1 generation of clover seedlings of transgenic plants and the corresponding salt of TP309 simultaneous handling of stress. When treating the base medium was 1 / 2MS. Salt treatment is 150mM NaCl, photographed before treatment, photographed 7 days after treatment, and then using the 1 / 2MS rehydration photographed 20 days, the survival rate statistics.

[0062] Firstly, TP309 and T 1 Generation ONAC103 overexpressing transgenic plants to salt stress treatment to salt stress analysis ONAC103 regulation case. Specific process is as follows: The rice seeds dehulled, sterilized by the method described hereinabove after sowing plants overexpressing transfected genes in 1 / 2MS medium containing 50mg / L hygromycin, TP309, sown in a solid 1 / 2MS culture groups. Be seed germination, the main root grow out of tissue culture flask, tissue culture was transferred to t...

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Abstract

Rice salt stress-resistance-related gene ONAC103 and its encoded protein and application relate to the technical fields of molecular biology and genetic engineering. A rice salt stress-resistant related gene ONAC103 (LOC_Os07g48450) is disclosed, and the nucleic acid sequence of the gene is shown in SEQ ID 1. ONAC103 is a gene related to salt stress resistance in rice. The relative expression of ONAC103 was significantly up-regulated under salt stress treatment. The salt tolerance of transgenic rice plants overexpressing ONAC103 was significantly higher than that of the control group. This gene can significantly improve the salt tolerance of rice. Therefore, the gene has important application value for screening salt-resistant rice varieties and improving rice yield. The cultivation of rice with enhanced resistance to salt stress in agricultural production is of great significance to energy and water conservation, utilization of saline-alkali land, and increase of grain production.

Description

Technical field [0001] The present invention relates to molecular biology, genetic engineering technology, in particular, involving rice resistant salt stress related gene onac103 and its encoded protein. Background technique [0002] Rice is one of the main food crops in the world, and its output is critical to food security. At the same time, rice can also be used as a model plant in plant research, and it is of great significance for the study of plant-salt stress mechanisms. The molecular biology mechanism of growth and development in rice and the regulation of adverse stress will not only help to understand the growth and development mechanism of rice, but also has important significance for rice yield. [0003] Although my country is vast, the country has a large number of people, but there are many people in my country, causing the per capita cultivated area inadequate, so that my country's can be severely used. At the same time, there is still a large area of ​​salt base ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8273
Inventor 陈亮赵立明李燕崔玉超郭迟鸣郭小玲
Owner XIAMEN UNIV
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