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Potassium ion channel protein KC1-D as well as encoding gene and application thereof

A technology of potassium ions and genes, applied to the potassium ion channel protein KC1-D and its coding gene and application field, to achieve the effect of increasing tolerance, increasing absorption capacity and absorption speed

Active Publication Date: 2016-04-27
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although increasing the amount of potassium fertilizer application can greatly increase crop yields, there are certain limitations to importing a large amount of potassium fertilizer both economically and in the long-term strategic sense.

Method used

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  • Potassium ion channel protein KC1-D as well as encoding gene and application thereof
  • Potassium ion channel protein KC1-D as well as encoding gene and application thereof
  • Potassium ion channel protein KC1-D as well as encoding gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, the acquisition of KC1-D protein and its coding gene

[0056] Mutants with low-potassium tolerance traits were obtained by screening for low-potassium stress on hundreds of thousands of Columbia ecotype Arabidopsis seedlings induced by EMS random mutagenesis. After sequencing, it was found that compared with the Colombian ecotype Arabidopsis, the mutant had a SNP mutation (mutation from G to A) in an open reading frame. The corresponding open reading frame in the mutant is shown in sequence 2 of the sequence listing, encoding the protein shown in sequence 1 of the sequence listing. The corresponding open reading frame in Arabidopsis thaliana ecotype is shown in sequence 4 of the sequence listing, and encodes the protein shown in sequence 3 of the sequence listing.

[0057] Compared with sequence 3, the difference of sequence 1 is only that the 322nd amino acid residue in sequence 3 is mutated from glycine to aspartic acid. Compared with sequence 4, the d...

Embodiment 2

[0060] Example 2, Functional verification of KC1-D protein

[0061] 1. Construction of recombinant plasmids

[0062] 1. Synthesize the double-stranded DNA molecule shown in sequence 2 of the sequence listing.

[0063] 2. Using the double-stranded DNA molecule obtained in step 1 as a template, a primer pair composed of Primer1 and Primer2 is used for PCR amplification to obtain a PCR amplification product, and a DNA fragment of about 2000 bp is recovered.

[0064] Primer1: 5'-GC TCTAGA ATGTCTACGACGACTACTGAGG-3';

[0065] Primer2: 5'-TC GAGCTC TTAGAAAATATATAAATGATCGTT-3'.

[0066] 3. Digest the DNA fragment obtained in step 2 with restriction endonucleases XbaI and SacI, and recover the digested product.

[0067] 4. Digest the Super1300 vector with restriction endonucleases XbaI and SacI, and recover the vector backbone of about 10,000 bp.

[0068] 5. Ligate the digested product of step 3 with the vector backbone of step 4 to obtain recombinant plasmid A. According to t...

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Abstract

The invention discloses potassium ion channel protein KC1-D as well as an encoding gene and an application thereof. The provided protein is named as KC1-D protein and is protein constituted by amino acid sequences in a sequence 1 in a sequence table. A gene (KC1-D gene) for encoding the KC1-D protein also belongs to the protection range of the invention. The invention further protects a method for culturing a transgenic plant. The KC1-D gene is introduced into a target plant, and the transgenic plant is obtained; the transgenic plant meets at least one phenotype from (d1) to (d3): (d1) the potassium ion absorbing ability is higher than that of the target plant; (d2) the potassium ion absorbing speed is higher than that of the target plant; (d3) the stress tolerance to the low-potassium environment is higher than that of the target plant. The potassium ion channel protein KC1-D has higher application value in improvement of crop molecules and has important theoretical significance and practical significance in culture of crops tolerant to low-potassium stress.

Description

technical field [0001] The invention relates to a potassium ion channel protein KC1-D and its coding gene and application. Background technique [0002] Potassium is one of the most abundant cations in plant cells and one of the three major nutrients necessary for plant growth. Potassium plays an irreplaceable role in many aspects of plant living cells, such as osmotic potential regulation, charge balance, sugar transport, protein synthesis, and the regulation of various enzyme activities in cells. Potassium plays an important role in plant growth and development. It participates in many physiological and biochemical metabolic processes of plants, and has an important impact on the normal growth, development and yield of plants. [0003] As a large agricultural country in my country, most of the cultivated land soil presents a potassium deficiency situation. However, my country's potash reserves are low, and potash fertilizers are seriously lacking, and more than half of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8271
Inventor 王毅武维华王雪萍陈丽梅
Owner CHINA AGRI UNIV
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