Eggplant potassium ion channel protein SmAKT1 as well as encoding gene and application thereof

A protein and coding technology, applied in the biological field, can solve problems such as research lag, and achieve the effect of enhancing tolerance and great application value

Active Publication Date: 2020-06-05
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on the response of eggplant to low potassium or salt stress is relatively

Method used

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  • Eggplant potassium ion channel protein SmAKT1 as well as encoding gene and application thereof
  • Eggplant potassium ion channel protein SmAKT1 as well as encoding gene and application thereof
  • Eggplant potassium ion channel protein SmAKT1 as well as encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1, the cloning of eggplant SmAKT1 gene

[0043] 1. Acquisition of plant material

[0044] The plant material used in this experiment was eggplant. Eggplants are planted at the Aerospace Breeding Base in Minhang District, Shanghai.

[0045] 2. Extraction of RNA

[0046] All the centrifuge tubes and tips of Ribonuclease (RNase) used in this experiment were purchased from Sangon Bioengineering (Shanghai) Co., Ltd. Mortars and pestles used were autoclaved. Total RNA was extracted using TaKaRa MiniBEST Universal RNA Extraction Kit (purchased from Baoriji Biotechnology (Beijing) Co., Ltd.). RNA quality was detected by agarose gel electrophoresis and NanoDrop2000 spectrophotometer.

[0047] 3. RNA reverse transcription into cDNA

[0048] Reverse Transcription Kit using PrimeScript TM II 1st Strand cDNA Synthesis Kit kit (purchased from Baori Medical Biotechnology (Beijing) Co., Ltd.).

[0049] 4. Cloning of target gene

[0050] Use primers

[0051] F1 (SEQ I...

Embodiment 2

[0054] Example 2, Sequence Information and Homology Analysis of Eggplant SmAKT1 Gene

[0055] The full-length CDS open reading frame sequence of the novel eggplant SmAKT1 gene of the present invention is 2655bp, and the detailed sequence is shown in SEQ ID NO.1. According to the CDS open reading frame sequence, the amino acid sequence of eggplant SmAKT1 is deduced, with a total of 884 amino acid residues. See the sequence shown in SEQ ID NO.2 for the detailed sequence.

[0056] The CDS open reading frame sequence of eggplant SmAKT1 and the amino acid sequence of its encoded protein were searched for nucleotide and protein homology in the Arabidopsis Genome Database using the BLAST program, and it was found that it was similar to the Arabidopsis AtAKT1 gene (AT2G26650.1 ) have 67.15% identity at the nucleotide level; at the amino acid level, they have 70.03% identity between them, such as figure 1 shown.

Embodiment 3

[0057] Example 3. Expression difference of SmAKT1 gene in different tissues of eggplant and its expression pattern under low potassium and salt stress

[0058] 1. Acquisition of plant material

[0059] Roots, stems, leaves, flowers, fruits, and sepals were taken from eggplant plants of 4-month seedling age. The samples were wrapped with aluminum platinum paper and put into liquid nitrogen immediately, and then transferred to -80°C ultra-low temperature freezer for storage until use. Three plants were taken from each sample, and the experiment was repeated three times.

[0060] Eggplant seedlings sown at the same time in the artificial climate culture room and with the same growth were selected, and when they grew to four leaves and one heart, they were treated with low potassium and salt stress respectively, and the leaves of the treated materials were taken at 0h, 6h, 12h, 24h, 48h and 7 days and root system. Three samples were taken at each time point, and each set of exp...

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Abstract

The invention discloses eggplant potassium ion channel protein SmAKT1 as well as an encoding gene and an application thereof. The protein SmAKT1 has the amino acid sequence shown in SEQ ID NO.2, and anucleotide sequence encoding the gene of the protein is shown in SEQ ID NO.1. The gene has expression in roots, stems, leaves, flowers, fruits and sepals, and the expression quantity in a root systemis notably higher than that in other tissue. Under low-potassium and salt stress treatment, the expression quantity of the gene in the leaves and the root system has the trend of first rising and then dropping. The gene function of the protein SmAKT1 is also identified, the gene SmAKT1 is imported in a target defect type yeast strain or an arabidopsis thaliana mutant, and the stress resistance ofthe gene to low-potassium stress and salt stress can be improved. A theoretical basis is provided for cultivation of the low-potassium stress and salt stress resistant eggplants with a gene engineering technology in the future, and the gene has great application value.

Description

technical field [0001] The invention relates to an eggplant potassium ion channel protein SmAKT1 and its coding gene and application, belonging to the field of biotechnology. Background technique [0002] Potassium is one of the three major nutrients (nitrogen, phosphorus, potassium) necessary for plant growth and development. Potassium ions are widely distributed in various tissues in plants, and are the most abundant cations in plants. They participate in various physiological processes in plants. The process plays a vital role in the normal growth and development of plants. [0003] my country is a large agricultural country, but due to the irrational cultivation and management modes of human beings such as fertilization, irrigation, and crop rotation, the balance of mineral ions in the soil has been broken, and most cultivated land soils are in a state of potassium deficiency and salinization. However, under salt stress, due to the excessive absorption of Na by plant ro...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/11C12N1/19C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8271C12N15/8273
Inventor 刘杨李静陈火英
Owner SHANGHAI JIAO TONG UNIV
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