Protein GhKT2 related to potassium ion absorption capacity of plant as well as coding gene and application thereof

A gene and plant technology, which is applied to the protein GhKT2 related to the ability of plants to absorb potassium ions and its encoding gene and application fields, can solve the problems of low potassium absorption ability and the like, 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 GhKT2 related to potassium ion absorption capacity of plant as well as coding gene and application thereof
  • Protein GhKT2 related to potassium ion absorption capacity of plant as well as coding gene and application thereof
  • Protein GhKT2 related to potassium ion absorption capacity of plant as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Discovery, expression analysis and subcellular localization of GhKT2 protein and its coding gene

[0038] 1. Discovery of GhKT2 protein and its coding gene

[0039] Use the database to search and compare related gene sequences of other species, splice 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 GhKT2 protein , as shown in sequence 1 of the sequence listing (composed of 792 amino acid residues). The gene encoding GhKT2 protein is named as GhKT2 gene, and the open reading frame of its cDNA is shown in sequence 2 of the sequence table (consisting of 2379 nucleotides).

[0040] In order to analyze the difference between the GhKT2 gene and the homologous gene sequences of other species, firstly, the multiple sequence alignment structure of the EBP1 sequence was constructed by using clustalX version1.83, and the default parameters of the software were used. According...

Embodiment 2

[0048] Embodiment 2, the acquisition of transgenic plants

[0049] 1. Construction of recombinant plasmids

[0050] 1. Extract the total RNA from the leaves of the cotton variety "Liaomian No. 17" and reverse transcribe it into cDNA.

[0051] 2. Using the cDNA obtained in step 1 as a template, perform PCR amplification with a primer pair composed of F1 and R1 to obtain a PCR amplification product.

[0052] F1: 5'-GC TCTAGA ATGGATCTTGAGTTTGGGAAGT-3'

[0053] R1: 5'-C GAGCTC TTACACCACATAAACCATGCCA-3'

[0054] 3. Digest the PCR amplification product obtained in step 2 with restriction endonucleases XbaI and SacI, and recover the digested product.

[0055] 4. The pBI121 vector was double-digested with restriction enzymes XbaI and SacI, and the vector backbone of about 12000 bp was recovered.

[0056] 5. Ligate the digested product of step 3 with the vector backbone of step 4 to obtain recombinant plasmid pBI121-GhKT2. The schematic diagram of the structure of the recombin...

Embodiment 3

[0074] Embodiment 3, identification of transgenic plants

[0075] MS medium formula:

[0076] Macroelements: 1.65g NH 4 NO 3 , 1.9g KNO 3 , 0.17g KH 2 PO 4 , 0.37 g m g SO 4 ·7H 2 O, 0.44 gCaCl 2 2H 2 O;

[0077] Trace elements: 22.3mg MnSO 4 4H 2 O, 8.6mg ZnSO 4 ·7H 2 O, 0.025mg CoCl 2 ·6H 2 O, 0.025mgCuSO 4 ·5H 2 O, 0.025mg Na 2 MoO 4 2H 2 O, 0.83mg KI, 6.2mg H 3 BO 3 ;

[0078] Iron salt: 27.8mg FeSO 4 ·7H 2 O, 37.3 mg Na 2 EDTA;

[0079] Dissolve macroelements, trace elements, iron salts and 9g of agar in water and dilute to 1L with water to obtain MS medium (potassium ion concentration is about 19.9mM).

[0080] Low Potassium Medium Formula:

[0081] 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;

[0082] Trace elements: same as those in MS medium.

[0083] Iron salt: the same as the iron salt in MS medium.

[0084] Dissolve macroelements, trace elements, iron salts and 9g agar ...

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Abstract

The invention discloses a protein GhKT2 related to potassium ion absorption capacity of a plant as well as a coding gene and application thereof. The protein provided by the invention is (a) or (b), wherein (a) is a protein composed of an amino acid sequence as shown in a sequence 1 in a sequence list, (b) is a protein formed by the way that substitution and / or deletion and / or addition of one or more than one amino acid residues are carried out on an amino acid sequence as shown in the sequence 1 and derivation related to low-potassium adversity stress resistance reaction of the plant, the potassium ion absorption capacity of the plant and development of the plant is carried out on the sequence 1. The invention discloses a new protein and gene, and after the gene is introduced into the plant, the low-potassium resistance of the plant can be obviously improved; the protein GhKT2 has a significant value on a new plant variety, especially a new cotton variety.

Description

technical field [0001] The invention relates to a protein GhKT2 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 aw...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
CPCC07K14/415C12N15/8261C12N15/8271
Inventor 田晓莉徐娟
Owner CHINA AGRI UNIV
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