Application of ebs1 gene in enhancing Arabidopsis resistance to bicarbonate stress

A bicarbonate, Arabidopsis technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problem of high phytotoxic effect, and achieve the effect of enhanced tolerance and robust root growth

Active Publication Date: 2021-12-03
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the poisonous effect of sodium bicarbonate on plants is also higher than that of high alkalinity stress

Method used

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  • Application of ebs1 gene in enhancing Arabidopsis resistance to bicarbonate stress
  • Application of ebs1 gene in enhancing Arabidopsis resistance to bicarbonate stress
  • Application of ebs1 gene in enhancing Arabidopsis resistance to bicarbonate stress

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Arabidopsis to NaHCO 3 , NaOAc dose-response

[0044] This embodiment analyzes Arabidopsis to NaHCO 3 , NaOAc dose response.

[0045] The specific method is as follows:

[0046] (1) Place the sterilized wild-type Arabidopsis Col-0 seeds on the MS medium (1% sucrose) containing the specified alkaline salt, and set the concentration range of 0-5 μM NaHCO 3 In the experimental group, a gradient was set for every 1.5 μM and for the NaOAc experimental group with a concentration range of 0-8 μM, a gradient was set for every 2 μM. After two days of vernalization at 4°C, the Arabidopsis seedlings were moved to the growth chamber for culture Cultivated in the box, Arabidopsis culture conditions are light intensity 150-200μmol·m -2 the s -2 , 60% humidity, 12 / 12h (light / dark) photoperiod.

[0047] (2) Take photos and record after 10 days of culture, measure root length with ImageJ, and statistically analyze the data by Student's t test or linear mixed model.

[0...

Embodiment 2

[0049] Example 2 RNA-Seq analysis of Arabidopsis in NaHCO 3 , Differential gene expression after NaOAc treatment

[0050] in NaHCO 3 , NaOAc treatment, the root and leaf samples were subjected to RNA sequencing, and then the differentially expressed genes were identified and analyzed.

[0051] The specific method is as follows:

[0052] (1) The seedlings grown for 10 days were transferred to 1 / 2MS medium containing or not containing sodium bicarbonate (3mM) and sodium acetate (4mM) (the treatment concentration was selected according to Example 1). Four days later, the roots and leaves of these seedlings were collected separately. A total of 120 seedlings (40 plants / plate, 3 plates / treatment) were collected for each genotype as a repetition.

[0053] (2) Use MiniBEST universal RNA Extraction Kit ( RNA Plant, Cat.#740949.50, TaKaRa; www.takarabio.com) to extract RNA;

[0054] (3) Illumina HiSeq 4000 is sequenced at 150bp paired ends; fine-tuning -0.38 is used to remove ad...

Embodiment 3

[0058] Example 3 accepts NaHCO 3 Deletion mutants of 13 genes specifically upregulated by NaHCO 3 Sensitivity test

[0059] The 13 gene T-DNA mutants were purchased from NASC (http: / / arabidopsis.info / BasicForm).

[0060] The specific experimental method is as follows:

[0061] (1) Plant 13 mutants and Col-0 in NaHCO-containing 3 (3mM) on the 1 / 2MS culture dish, the cultivation method is consistent with embodiment 1;

[0062] (2) After 10 days, take pictures to record the growth status of the plants, and use ImageJ to measure the root length.

[0063] Result: in NaHCO 3 Sensitivity verification experiments screened out the mutant lines of AT5G07330 (Salk_104370C), AT3G05950 (Salk_038626C) and AT4G17680 (Sail_420_E12) in the sodium bicarbonate treatment environment with poor plant growth status, weak root system, and sensitive to stress. ( Figure 5 A).

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Abstract

The invention discloses the application of EBS1 gene in enhancing Arabidopsis thaliana resistance to bicarbonate stress. The nucleotide sequence of the protein coding region of the EBS1 gene is shown in SEQ ID No.1. The present invention identifies and screens a gene EBS1 resistant to bicarbonate stress, and provides a new application of EBS1 as a positive regulator in regulating Arabidopsis bicarbonate resistance and improving planting in saline-alkali land, for the cultivation of bicarbonate-tolerant plants Provide evidence.

Description

technical field [0001] The invention relates to the technical field of biotechnology, and mainly relates to the application of the EBS1 gene in enhancing the ability of Arabidopsis to resist bicarbonate stress. Background technique [0002] The saline-alkali land area in the world reaches 950 million hectares, accounting for about one-third of the land area, while the saline-alkali land area in my country is nearly 100 million hectares. Plants in alkaline soils are under multiple pressures, such as osmotic stress, nutrient deficiency, ion poisoning, and high pH stress. Wait. The lack of nutrients, high pH, ​​high salinity, and rich bicarbonate in the soil affect the normal metabolism of plants. Therefore, deepening the research on the mechanism of plant salt-alkali tolerance will help the efficient use of soil resources and the development of agricultural production. [0003] It has been found that plants will regulate a series of physiological activities such as enhancing t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82C07K14/415A01H5/00A01H6/20
CPCC07K14/415C12N15/8273
Inventor 吴莹崔富强李潇潇杨一帆
Owner ZHEJIANG FORESTRY UNIVERSITY
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