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Use of Arabidopsis thaliana proton pump gene AtNHX1 for promoting plant rootage absorbing potassium capability

A plant root system and proton pump technology, applied in the fields of application, plant peptides, genetic engineering, etc., to achieve the effect of increasing the potassium content of plants and improving the potassium absorption capacity of transformed plant roots

Inactive Publication Date: 2006-12-27
黑龙江省烟草科学研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no report about the Arabidopsis proton pump gene AtNHX1 improving the potassium uptake ability of plant roots

Method used

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  • Use of Arabidopsis thaliana proton pump gene AtNHX1 for promoting plant rootage absorbing potassium capability
  • Use of Arabidopsis thaliana proton pump gene AtNHX1 for promoting plant rootage absorbing potassium capability
  • Use of Arabidopsis thaliana proton pump gene AtNHX1 for promoting plant rootage absorbing potassium capability

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Designing primers related to AtNHX1 gene

[0036] 1. The primer sequence for amplifying the proton pump gene (AtNHX1) of Arabidopsis:

[0037] Upstream: 5'-AAAGGATCCAACATGTTGGATTCTCTAGTGTCGAAAC-3' SEQ ID No.3

[0038] Downstream: 5'-AAAGAGCTCTCAAGCCTTACTAAAGATCAGGAGGG-3' SEQ ID No.4

[0039] Restriction sites for BamHI and SacI were designed at the 5' end of the primers;

[0040] 2. Primer sequences for PCR detection of transformed materials:

[0041] AtNHX1-Z: 5′-CGGTCTGATAAGTGCGTATG-3′SEQ ID No.5

[0042] AtNHX1-F: 5′-GTTCTGGTGCGGTAATAGGT-3′SEQ ID No.6

[0043] The length of the amplified fragment is 650bp;

[0044] 3. Fluorescent quantitative PCR amplification primers for mRNA transformed tobacco

[0045] AtNHX1-SA: 5′-CCTATTACCGCACCAGAACG-3′SEQ ID No.7

[0046] AtNHX1-SB: 5′-GGTCGCATGAAGGAGTCATC-3′SEQ ID No.8

Embodiment 2

[0047] Example 2: Arabidopsis total RNA extraction and purification

[0048] Take 1 g of Arabidopsis thaliana root, add liquid nitrogen, grind it into powder in a mortar, put it into a 1.5ml microcentrifuge tube; extract total RNA with Promega total RNA extraction kit, operate according to the instructions, and judge the extracted RNA by gel electrophoresis quality.

Embodiment 3

[0049] Embodiment 3: PCR synthesis of AtNHX1 gene

[0050] The Arabidopsis thaliana total RNA extract obtained in Example 2 was used as a template, and Oligo DT was used as a primer to synthesize the first strand of cDNA by reverse transcription. Using the reverse transcription product as a template, the upstream and downstream primers of the reading frame of the Arabidopsis proton pump gene (AtNHX1) and Ex-taq DNA polymerase were used for PCR amplification under conventional reaction conditions.

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Abstract

This invention discovered for the first time and exposed that the proton pump gene of Arabidopsis thaliana has the function of increasing the kalium absorption capability of root system. This invention also provides the construction method of plant expression vector that contains this gene and the approach that can increas the content of kalium in plant. The benefits of this invention is that it discovered for the first time that the proton pump gene of arabidopsis thaliana has the function of increasing the kalium absorption capability of root system, as well as provides the construction method of plant expression vector that contains this gene and the approach that can increas the content of kalium in plant. This invention can resolve the technical problem that the content of kalium in tobacco is too low. This problem restricts the development of tobacco technology in our country.

Description

technical field [0001] The invention relates to the application of proton pump gene AtNHX1 of Arabidopsis thaliana to improve the ability of plant root system to absorb potassium, and belongs to the field of plant molecular biology. Background technique [0002] Potassium is an activator of various enzymes in tobacco plants, and there are more than 50 enzymes in cells or completely dependent on K + or by K + activation of K + It can regulate many physiological functions of plants, such as enhancing photosynthesis, enhancing material synthesis and transport in plants, improving plant resistance, maintaining cell turgor, regulating stomatal movement, and increasing CO 2 increase the assimilation rate of photosynthesis and photosynthetic products, and significantly increase the sugar content in the tobacco plant. The increase in sugar content can not only increase the osmotic potential of cells, enhance drought resistance, but also lower the freezing point, reduce frost damag...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82
Inventor 郭兆奎姚泉洪万秀清颜培强李丽杰
Owner 黑龙江省烟草科学研究所
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