Application of OsCIPK23 protein to cultivation of low-potassium stress-resistant plant

A low-potassium, plant-resistant technology, applied in the fields of application, plant products, plant genetic improvement, etc., can solve problems such as high price and potassium environmental hazards

Inactive Publication Date: 2013-12-18
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Potassium resources in my country are scarce, mainly rely on imports, and the price is expensive. ...

Method used

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  • Application of OsCIPK23 protein to cultivation of low-potassium stress-resistant plant
  • Application of OsCIPK23 protein to cultivation of low-potassium stress-resistant plant
  • Application of OsCIPK23 protein to cultivation of low-potassium stress-resistant plant

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the acquisition of OsCIPK23 protein and its coding gene

[0038] Obtain a DNA sequence (derived from the rice variety "Nipponbare") on the TIGR website (http: / / rice.plantbiology.msu.edu), which encodes the protein shown in Sequence 1 of the Sequence Listing.

[0039] The protein shown in Sequence 1 of the Sequence Listing was named OsCIPK23 protein (composed of 450 amino acid residues). The gene encoding OsCIPK23 protein is named OsCIPK23 gene, its genome sequence has 14 exons and 13 introns, and the open reading frame in its cDNA sequence is shown in sequence 2 of the sequence listing.

Embodiment 2

[0040] Embodiment 2, detection of transgenic Arabidopsis plant phenotype

[0041] 1. Construction of overexpression vector

[0042] 1. Synthesize the double-stranded DNA molecule shown in sequence 2 of the sequence listing.

[0043] 2. Using the double-stranded DNA molecule synthesized in step 1 as a template, perform PCR amplification with a primer pair composed of OsCIPK23-UN1301F and OsCIPK23-UN1301R to obtain a PCR amplification product.

[0044] OsCIPK23-UN1301F: 5'–TT GGATCC ATGAGCGTGTCGGGCGGGA-3';

[0045] OsCIPK23-UN1301R: 5'–TT GGTACC TCACGGTGACCTCCGATGCT-3'.

[0046] 3. Digest the PCR amplified product in step 2 with restriction endonucleases Bam HI and Kpn I, and recover the digested product.

[0047] 4. Digest the 1301-Ubiq plasmid with restriction endonucleases Bam HI and Kpn I to recover the vector backbone (about 11800bp).

[0048] 5. Ligate the digested product of step 3 with the vector backbone of step 4 to obtain a recombinant plasmid. According to t...

Embodiment 3

[0081] Embodiment 3, transgenic rice plant phenotype detection

[0082] 1. RNAi vector construction

[0083] 1. Synthesize the double-stranded DNA molecule shown in sequence 2 of the sequence listing.

[0084] 2. Using the double-stranded DNA molecule synthesized in step 1 as a template, perform PCR amplification with a primer pair composed of OsCIPK23-pTCK303F and OsCIPK23-pTCK303R to obtain a PCR amplification product.

[0085] OsCIPK23-pTCK303F: 5'-GG GGTACCACTAGT ACGCTGTCGATTACTGTCA-3';

[0086] OsCIPK23-pTCK303R: 5'-CG GGATCCGAGCTC TTGGAGGCTGATATCCC-3'.

[0087] 3. The PCR amplified product of step 2 was double-digested with restriction enzymes SpeI and SacI, and the digested product was recovered.

[0088] 4. Digest the pTCK303 plasmid with restriction enzymes SpeI and SacI, and recover the vector backbone (about 11800bp).

[0089] 5. Ligate the digested product of step 3 with the vector backbone of step 4 to obtain a recombinant plasmid.

[0090] 6. Double-dige...

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Abstract

The invention discloses application of OsCIPK23 protein to cultivation of a low-potassium stress-resistant plant. The invention provides a method of culturing a transgenic plant. The method comprises the following steps: an encoding gene of the OsCIPK23 protein is induced into a target plant, so that the transgenic plant is obtained. The low-potassium stress-resistant ability of the transgenic plant is higher than that of the target plant, and/or the potassium ion absorbility of the transgenic plant is higher than that of the target plant. The application can be used for improving low-potassium resistance of the plant and provides a guarantee for cultivating a new paddy rice variety suitable for soil deficient in potassium.

Description

technical field [0001] The invention relates to the application of OsCIPK23 protein in cultivating plants resistant to low potassium stress. Background technique [0002] Potassium is one of the three major nutrients (nitrogen, phosphorus, potassium) necessary for plant growth and development, and is also the most abundant monovalent cation in plants. Many plants have a large demand for potassium, and the potassium content in some plants can be as high as 5%-10%. Potassium exists in plants in the form of soluble inorganic salts or ions. It has the characteristics of wide distribution, high content, and strong mobility. It is mainly concentrated in the most vigorous parts of life activities (usually preferentially distributed in buds, young leaves, upper stems and root tips) and other vigorously growing organs and tissues). [0003] Potassium does not directly participate in the formation of any organic substances, but participates in the regulation of enzyme activity, and ...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/84A01H5/00
Inventor 王毅武维华李娟龙雨
Owner CHINA AGRI UNIV
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