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Low kalium resisting associated protein TaPR1 and coding protein and application thereof

A low-potassium and protein-resistant technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of inability to maintain soil potassium and other nutrients, reduce fertilizer utilization rate, and unbalanced potassium content in farmland, etc.

Active Publication Date: 2020-01-31
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the increasing demand for crop yield, my country's soil has experienced soil potassium deficiency caused by partial application of nitrogen and phosphorus fertilizers, and now it has changed to a large amount of application of three chemical fertilizers. The amount of potassium taken away is also increasing, and the amount of potassium applied to the soil still cannot maintain the balance of soil potassium and other nutrients, resulting in an imbalance of potassium content in the farmland. This imbalance of nutrients will reduce the utilization rate of fertilizers and become a limit to crop yield. Improvement factors (Xia Le, 2016; Sun Aiwen et al., 2009; Jin Jiyun, 2005; Sun Haifeng et al., 2018)

Method used

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  • Low kalium resisting associated protein TaPR1 and coding protein and application thereof
  • Low kalium resisting associated protein TaPR1 and coding protein and application thereof
  • Low kalium resisting associated protein TaPR1 and coding protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Cloning of embodiment 1, TaPR1

[0056] 1. Cloning of TaPR1

[0057] The roots of common wheat (Triticum aestivum L.) (variety Aikang 58) grown in hydroponics for about 5 days were taken from the roots of the three-leaf stage seedlings, quickly frozen in liquid nitrogen, and stored at -80°C for later use.

[0058] The total RNA of wheat seedling roots was extracted by Trizol method (TransGEN Biotech), and cDNA was synthesized by PrimerScriptTM RT reagent Kit with gDNAEraser kit (TaKaRa).

[0059] Amplification system (20μL):

[0060]

[0061]

[0062] Reaction procedure:

[0063]

[0064] PCR products were detected by 1.0% agarose gel electrophoresis.

[0065] The gene shown in Sequence 2 in the Sequence Listing is named TaPR1 gene, wherein its coding gene is the nucleotides at positions 80-2218 at the 5' end of Sequence 2 in the Sequence Listing, and the protein encoded by the gene is named TaPR1 protein, the amino acid sequence of the protein is sequence ...

Embodiment 2

[0067] Example 2, real-time fluorescent quantitative PCR analysis of the expression characteristics of TaPR1

[0068] 1. Wheat material processing

[0069] a. At the wheat filling stage, take the wheat roots, stems, young leaves, old leaves and grains grown in the field, put them into liquid nitrogen for quick freezing, and store them at -80°C for later use.

[0070] B, to the wheat seedling of three-leaf stage, carry out following processing:

[0071] (1) Low-potassium treatment: place wheat seedlings in culture solution with a concentration of 0.1 mM potassium ions, and culture them under light for 1 hour, 3 hours, 6 hours, 9 hours, 12 hours, and 24 hours. Quick-frozen and stored at -80°C for later use.

[0072] (2) Treatment of the control: The roots of wheat seedlings without any treatment were directly frozen at -80° C. as a control.

[0073] 2. Extraction of RNA

[0074] Total RNA was extracted from wheat roots, stems, young leaves, old leaves and grains grown in fie...

Embodiment 3

[0091] Example 3, TaPR1 subcellular localization analysis

[0092] 1. Construct 35S-GFP vector:

[0093] A primer pair (TaPR1-1F and TaPR1-1R) was designed according to the sequence of the TaPR1 gene, and BamHI and SalI restriction sites were introduced at the ends of the primers respectively.

[0094] TaPR1-1F: 5'-CGC GGATCC ATGGATCTCGAGGCGGCAGCT-3';

[0095] TaPR1-1R: 5'-ACGC GTC GAC AACCTTGTACAACATGCCG-3'.

[0096] The target gene PCR product and 35S-GFP expression vector were double-digested with Thermo Fisher enzymes BamHI and SalI, respectively.

[0097] The double enzyme digestion system (20μL) is as follows:

[0098]

[0099] Use a PCR machine to incubate at 37°C for 70 minutes, and inactivate the enzyme according to the requirements of the instructions for subsequent experiments.

[0100] Gel recovery was performed on the double-digested products, and TAKARA T4 ligase was used for ligation. The molar ratio of the target fragment:carrier was 10:1.

[0101] ...

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Abstract

The invention discloses low kalium resisting associated protein TaPR1 and a coding protein and application thereof. The protein TaPR1 provided by the invention is protein of which the amino acid sequence is shown as a sequence 1 in a sequence table, and the coding gene is DNA of which the nucleotide sequence is a sequence 2 in the sequence table. The protein and the gene provided by the inventionprovide a foundation for manual control of expression of a potassium ion absorption related gene, and can exert important effects on breeding of low kalium resisting plants and potassium ion absorption strengthening plants.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a low potassium resistance-related protein TaPR1 and its coding gene and application. Background technique [0002] Nitrogen, phosphorus and potassium are the three elements of fertilizers for crop growth. Since the mid-1950s, they have been applied in my country's agricultural production (Shen Shanmin et al., 1998). The use of chemical fertilizers has ensured my country's food security. With the increasing demand for crop yield, my country's soil has experienced soil potassium deficiency caused by partial application of nitrogen and phosphorus fertilizers, and now it has changed to a large amount of application of three chemical fertilizers. The amount of potassium taken away is also increasing, and the amount of potassium applied to the soil still cannot maintain the balance of soil potassium and other nutrients, resulting in an imbalance of potassium content in the farmland. This i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/81C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8271C12N15/81Y02P60/21
Inventor 程西永毕惠惠许海霞刘晓丹张旭睿
Owner HENAN AGRICULTURAL UNIVERSITY
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