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Gene for promoting plants to absorb zinc ions, amino acid sequence, expression vector and application

A technology for plant absorption and expression vectors, applied in the field of plant biology, can solve the problems of undiscovered genes and insufficient research, and achieve the effect of increasing the content

Inactive Publication Date: 2017-05-31
INNER MONGOLIA HESHENG ECOLOGICAL SILVICULTURE CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the current research on the genes that promote the absorption of zinc ions by plants is not thorough enough, and new genes that affect the promotion of zinc ion absorption by plants have yet to be discovered.

Method used

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  • Gene for promoting plants to absorb zinc ions, amino acid sequence, expression vector and application
  • Gene for promoting plants to absorb zinc ions, amino acid sequence, expression vector and application
  • Gene for promoting plants to absorb zinc ions, amino acid sequence, expression vector and application

Examples

Experimental program
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Effect test

Embodiment 1

[0040] A gene that promotes zinc ion absorption by plants, the sequence of the gene is shown in SEQ ID NO:1. Correspondingly, an amino acid sequence encoded by the gene is shown in SEQ ID NO:8.

[0041] A first primer set for identifying the gene mutation, a second primer set and a third primer set, the first primer set includes an upstream primer F1 and a downstream primer R1, the sequence of the upstream primer F1 is shown in SEQ ID NO: 2 The sequence of the downstream primer R1 is shown in SEQ ID NO: 3; the second primer set includes upstream primer F2 and downstream primer R2, the sequence of the upstream primer F2 is shown in SEQ ID NO: 4, the downstream The sequence of primer R2 is shown in SEQ ID NO: 5; the third primer set includes upstream primer Actin F and downstream primer Actin R, the sequence of the upstream primer Actin F is shown in SEQ ID NO: 6, and the downstream primer Actin The sequence of R is shown in SEQ ID NO:7.

Embodiment 2

[0043] This example is the functional identification of the ZNE gene.

[0044] The frozen zinc ion-deficient yeast was streaked onto YPDA solid medium and cultured upside down at 30°C until monoclonal colonies grew. Then the monoclonal colony was transferred to 3ml YPDA medium and cultured at 250rpm at 30°C for 8-12h. Take 5ul of the culture solution and expand it into 50ml of YPDA medium at 30°C and 250rpm to continue culturing until the OD600 reaches 0.15-0.3. Centrifuge at 700g for 5min at room temperature, discard the supernatant, and resuspend the cells with 100ml of fresh YPDA. Continue culturing at 30°C and 250 rpm until the OD600 reaches 0.4-0.5. Divide the medium into two 50ml sterile tubes, centrifuge at 700g for 5min at room temperature, discard the supernatant, and add 30ml ultrapure water to each tube to resuspend the bacteria. Centrifuge at 700g for 5min at room temperature, discard the supernatant, and add 1.5ml 1.1X TE / LiAc to each tube to resuspend the bacte...

Embodiment 3

[0048] This example is a verification test of the function of the ZNE gene in Arabidopsis plants.

[0049] The experimental materials in this example are mainly the Arabidopsis wild-type material Col-0 containing the ZNE gene, and the deletion homozygous mutants zne1-1 and zne1-2 of the ZNE gene obtained by inserting T-DNA, wherein, zne1-1 is a deletion homozygous mutant obtained by inserting chr 3 2625013bp coding region sequence of Arabidopsis wild-type gene into T-DNA; zne1-2 is obtained by inserting chr 3 2626125bp coding region sequence of Arabidopsis wild-type gene into T-DNA Deletion homozygous mutants. Both zne1-1 and zne1-2 belong to the ZNE gene deficiency type because of the insertion of T-DNA in the gene, which leads to the destruction of the function of the gene.

[0050] 1. Verify the genotypes of Col-0, zne1-1 and zne1-2.

[0051] Col-0, zne1-1 and zne1-2 were respectively subjected to PCR amplification using the primer set in Example 1 to verify their genotyp...

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Abstract

The invention provides a gene for promoting plants to absorb zinc ions, an amino acid sequence, an expression vector and an application. The gene for promoting the plants to absorb the zinc ions, the amino acid sequence and the expression vector have very important theoretical and practical significance in improvement and increase of the content of the zinc ions of the plants and accelerated cultivation of high zinc-containing crops and new pasture grass varieties.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, and in particular relates to a gene, amino acid sequence, expression vector and application for promoting zinc ion absorption by plants. Background technique [0002] Zinc is an essential mineral element for plant growth, a cofactor for various enzymes in organisms, and participates in various biochemical metabolic reactions of organisms. The lack of zinc will seriously affect the physiological and metabolic process of plants, so it is very important to strictly control the balance of zinc in plants. Zinc deficiency stress exists in about 30% of the world's soil including China. Investigations show that about 45.7% of the arable land in my country has soil available zinc lower than the critical level of 0.5mg / kg. [0003] In the 1940s, Stout, Arnon, Skoog, and Hoagland confirmed through experiments that zinc is an essential trace mineral element in the growth and development of higher plants....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12Q1/68C12N15/11C07K14/415C12N15/82A01H5/00
CPCC07K14/415C12N15/8243C12Q1/6895C12Q2600/156
Inventor 赵泉胜
Owner INNER MONGOLIA HESHENG ECOLOGICAL SILVICULTURE CO LTD
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