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Tobacco KUP2 gene and applications thereof

A gene and tobacco technology, applied in the tobacco KUP2 gene and application field, can solve the problems of large potassium consumption, reduced potassium ion concentration and the like

Active Publication Date: 2019-04-02
GUIZHOU TOBACCO SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Existing technology studies on potassium transporter genes are extensive in the model plant Arabidopsis. For example, studies have shown that Arabidopsis mutant kup6 has a lower survival rate under drought stress than wild-type plants, while 35S::KUP6 overexpression However, the drought tolerance of transgenic plants was significantly enhanced (Shabala et al., 2007); while the KUP7 mutant seedlings showed a low-potassium-sensitive phenotype of yellowing leaves under low-potassium stress, and the potassium ion content in the roots was significantly The loss of KUP7 gene function decreased the ability of Arabidopsis seedlings to absorb potassium under low potassium conditions, and decreased the concentration of potassium ions in xylem sap (Min et al., 2016)
[0004] Tobacco is a crop that consumes a lot of potassium. The potassium content of tobacco leaves is an important indicator to measure the quality of tobacco leaves. At present, there are few studies on potassium transporter genes in tobacco, and the function of tobacco KUP gene is unknown.

Method used

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  • Tobacco KUP2 gene and applications thereof
  • Tobacco KUP2 gene and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Cloning of Example 1 Tobacco KUP2 Gene

[0032] Get 0.5g of tobacco (tobacco variety is K326) fresh leaves, use the Trizol method to extract the total RNA of tobacco cells, then use the cDNA synthesis kit of TaKaRa company to synthesize cDNA, and further use Primer5.0 software to design and obtain primers through artificial optimization. The primers include a forward primer and a reverse primer, the nucleotide sequence of the forward primer is: 5'-ATGGATATTGAAAGTTGGG-3'; the nucleotide sequence of the reverse primer is 5'-TTATACATGGTAAATCATTC-3', Use the synthesized cDNA as a template for PCR amplification. The PCR amplification system is 20 μL, including 10 μL of Premix ExTaq, 0.5 μL of 10 μM forward primer, 0.5 μL of 10 μM reverse primer, 1 μL of tobacco cell cDNA, ddH 2 O 8 μL; the reaction program of the PCR amplification is: pre-denaturation at 95° C. for 5 minutes; denaturation at 95° C. for 30 seconds; annealing at 55° C. for 30 seconds; extension at 72° C. for 2...

Embodiment 2

[0034] Example 2 Biological Functional Analysis of Tobacco KUP2 Gene

[0035] 1. Purpose of the experiment

[0036] The biological function of tobacco KUP2 gene was verified by yeast functional complementation experiment.

[0037] 2. Experimental method

[0038] The potassium uptake-deficient yeast mutant strain R5421 was used as the recipient strain. Strain R5421 can be found in Maathuis F J Mand Sanders D 1996 Mechanisms of potassium absorption by higher plantroots. Physiol. Plant. 96, 158–168.

[0039] The T-vector connected with the tobacco KUP2 gene of Example 1 and the expression vector P416 (yeast free type shuttle expression vector, TEF constitutive promoter, CYC1 terminator, CEN6ARSH4 origin of replication, the screening marker in yeast is URA3, screening in Escherichia coli Marked as Amp. Carrier P416 can be found in Functional Expression of aω-3Fatty Acid Desaturase Gene from Glycine max in Saccharomyces cerevisiae) to carry out double enzyme digestion respective...

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Abstract

The present invention relates to a tobacco KUP2 gene and applications thereof. The sequences of the tobacco KUP2 gene and a protein encoded by the gene are respectively shown in SEQ ID NO:1 and SEQ IDNO:2. The KUP2 gene is obtained by cloning from tobacco for the first time, and the biological functions of the gene are verified by a yeast functional complementation experiment. The tobacco KUP2 gene has the function of promoting potassium ion absorption and transport.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to tobacco KUP2 gene and its application. Background technique [0002] Potassium transporter is a carrier protein that can transport potassium ions into cells when the external potassium ion concentration is extremely low. Potassium transporters are usually some transmembrane proteins, which complete the transmembrane transport of potassium ions through conformational changes. It is an active transport process that requires ATP to provide energy, usually K + / H + or K + / Na + Collaborative transport. [0003] Existing technology studies on potassium transporter genes are extensive in the model plant Arabidopsis thaliana. For example, studies have shown that Arabidopsis mutant kup6 has a lower survival rate under drought stress than wild-type plants, while 35S::KUP6 overexpression The drought tolerance of transgenic plants was significantly enhanced (Shabala et al.,...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/81C12N1/19C12N15/82A01H5/00A01H6/82A01H6/20
CPCC07K14/415C12N15/81C12N15/8243
Inventor 张洁任学良李立芹鲁黎明王仁刚王自力
Owner GUIZHOU TOBACCO SCI RES INST
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