Protein related to plant potassium ion absorption capacity, and coding gent and application thereof

A protein and gene technology, applied in plant genetic improvement, botanical equipment and methods, applications, etc., can solve the problem of low potassium absorption capacity, and achieve the effect of improving low potassium tolerance

Active Publication Date: 2015-04-15
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, cotton has a low ability to absorb potassium in the soil, so cotton is more sensitive to low potassium in the soil than other field crops

Method used

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  • Protein related to plant potassium ion absorption capacity, and coding gent and application thereof
  • Protein related to plant potassium ion absorption capacity, and coding gent and application thereof
  • Protein related to plant potassium ion absorption capacity, and coding gent and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1. Discovery, expression analysis and subcellular localization of GhAKT1 protein and its coding gene

[0045] 1. Discovery of GhAKT1 protein and its coding gene

[0046] Use the database to search and compare related gene sequences of other species, assemble and verify the EST sequence of cotton, and perform 5'RACE and 3'RACE, obtain a new protein from the cotton variety "Liaomian 17", and name it GhAKT1 protein , as shown in Sequence 1 of the Sequence Listing (composed of 875 amino acid residues). The gene encoding GhAKT1 protein was named as GhAKT1 gene, and the open reading frame of its cDNA is shown in sequence 2 of the sequence table (consisting of 2628 nucleotides).

[0047] In order to analyze the difference between the GhAKT1 gene and the homologous gene sequences of other species, firstly, the multiple sequence alignment structure of the EBP1 sequence was constructed using the clustalX version1.83, using the default parameters of the software. Accordi...

Embodiment 2

[0055] Embodiment 2, the acquisition of transgenic plants

[0056] 1. Construction of recombinant plasmids

[0057] 1. Extract the total RNA from the leaves of the cotton variety "Liaomian No. 17" and reverse transcribe it into cDNA.

[0058] 2. Using the cDNA in step 1 as a template, perform PCR amplification with a primer pair composed of F1 and R1 to obtain a PCR amplification product.

[0059] F1: 5'-GC TCTAGA ATGTTTCGAGGGTCAGTACTAT-3'

[0060] R1: 5'-GG GGTACC TTAAGGGTTTTGGGTGTCATTA-3'

[0061] 3. The PCR amplified product of step 2 was double-digested with restriction endonucleases XbaI and KpnI, and the digested product was recovered.

[0062] 4. Digest the vector pBIB with restriction endonucleases XbaI and KpnI to recover a vector backbone of about 10000 bp.

[0063] 5. Ligate the digested product of step 3 with the vector backbone of step 4 to obtain the recombinant plasmid PBIB-GhAKT1. The schematic diagram of the structure of the recombinant plasmid PBIB-G...

Embodiment 3

[0091] Embodiment 3, identification of transgenic plants

[0092] MS medium formula (potassium ion concentration is about 19.9mM):

[0093] Macroelements: 1.65g NH 4 NO 3 , 1.9g KNO 3 , 0.17g KH 2 PO 4 , 0.37g MgSO 4 ·7H 2 O, 0.44g CaCl 2 2H 2 O;

[0094] Trace elements: 22.3mg MnSO 4 4H 2 O, 8.6mg ZnSO 4 ·7H 2 O, 0.025mg CoCl 2 ·6H 2 O, 0.025mg CuSO 4 ·5H 2 O, 0.025mg Na 2 MoO 4 2H 2 O, 0.83mg KI, 6.2mg H 3 BO 3 ;

[0095] Iron salt: 27.8mg FeSO 4 ·7H 2 O, 37.3 mg Na 2 EDTA;

[0096] Dissolve macroelements, trace elements, iron salts and 9 g of agar in water and dilute to 1 L with water to obtain MS medium. Low Potassium Medium Formula:

[0097] Macroelements: 2.3g NH 4 NO 3 , 0.37g MgSO 4 ·7H 2 O, 0.144g NH 4 h 2 PO 4 , 0.44g CaCl 2 2H 2 O;

[0098] Trace elements: same as those in MS medium.

[0099] Iron salt: the same as the iron salt in MS medium.

[0100] Dissolve macroelements, trace elements, iron salts and 9g agar in water and ...

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Abstract

The invention discloses a protein related to plant potassium ion absorption capacity, and a coding gent and an application thereof. The protein is obtained form a cotton variety Liaomian No.17, and is named as GhAKT1 protein. The protein is the following (a) or (b) or (c) or (d): (a) a protein composed of an amino acid sequence represented by a sequence 1; (b) a protein derived from the sequence 1 and related to plant low potassium stress tolerance, wherein the protein is obtained by that the sequence 1 is subjected to substitution and / or deletion and / or addition of one or more amino acid residues; (c) a protein derived from the sequence 1 and related to plant potassium absorption capacity, wherein the protein is obtained by that the sequence 1 is subjected to substitution and / or deletion and / or addition of one or more amino acid residues; and (d) a protein derived from the sequence 1 and related to plant development, wherein the protein is obtained by that the sequence 1 is subjected to substitution and / or deletion and / or addition of one or more amino acid residues. The protein, gene, and application provided by the invention have important values for plant novel varieties and especially cotton varieties.

Description

technical field [0001] The invention relates to a protein related to the ability of plants to absorb potassium ions, its coding gene and application. Background technique [0002] Potassium is one of the three essential nutrients for plant growth, and is closely related to enzyme activity, protein synthesis, photosynthesis, oil synthesis, stomatal movement, ion balance and stress resistance. Potassium nutrient deficiency leads to slow plant growth, short plant and poor root development or even rot, and reduced stress resistance and disease resistance. [0003] In recent years, with the promotion of high-yield varieties in my country, the increase in the application of nitrogen and phosphorus fertilizers, and the improvement of the quality requirements of agricultural products, the scope of potassium-deficient soil in my country has gradually expanded. 1 / 4 to 1 / 3 of China's arable land is deficient in potassium or severely deficient in potassium. The potassium taken away by ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N1/21C12R1/01
Inventor 田晓莉徐娟
Owner CHINA AGRI UNIV
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