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Corn arsenic stress resistant gene ZmAsR2 and intramolecular SNP label and application thereof

A technology of resistance gene, corn, applied in the direction of DNA/RNA fragment, application, genetic engineering, etc., can solve the problems of lateral root damage, reduce crop yield and so on

Inactive Publication Date: 2020-03-20
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, high concentrations of arsenic can also have adverse effects on plant growth, such as causing damage to lateral roots and inhibiting water absorption, thereby reducing crop yields

Method used

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  • Corn arsenic stress resistant gene ZmAsR2 and intramolecular SNP label and application thereof
  • Corn arsenic stress resistant gene ZmAsR2 and intramolecular SNP label and application thereof
  • Corn arsenic stress resistant gene ZmAsR2 and intramolecular SNP label and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Discovery, identification and cloning of arsenic stress resistance gene ZmAsR2 in maize

[0028] The material used in this study is an associated population composed of 350 representative inbred lines, of which 151 are from temperate regions, 79 are from tropical and subtropical regions, and they were planted in Yongcheng (YC) and Yuanyuan in Henan, China in 2017. In Yanghe Henan Agricultural University Base (YY), in each location, the associated populations were designed with completely randomized blocks, with three repetitions. The row length of each plot is 3 meters, the plant spacing is 0.22 meters, and the row spacing is 0.67 meters. The final planting density is 67,500 plants / ha.

[0029] The mature ears of the associated populations were harvested to determine the accumulation and distribution of arsenic in cobs and bracts. The cobs and bracts of each inbred line in each environment were collected together and dried naturally. The dried cobs and bract...

Embodiment 2

[0041] Example 2: Maize arsenic stress resistance gene ZmAsR2 Intramolecular SNP marker development

[0042] 1. Select the target sequence: select the target sequence containing the SNP site in gramene http: / / ensembl.gramene.org / Zea_mays / Info / Index);

[0043] 2. Design specific primers: design primers in dCAPS Finder 2.0 (http: / / helix.wustl.edu / dcaps / dcaps.html) containing the sequences of 25 nt upstream and downstream of the SNP site, and then insert the 25 nt sequences on the left side of the SNP site 4 bases G are replaced by A, such as image 3 As shown, the enzyme-cleaved recognition site AAGCTT of HindIII appears in the material resistant to arsenic stress.

Embodiment 3

[0044] Example 3: Maize arsenic stress resistance gene ZmAsR2 Application in Maize Breeding for Heavy Metal Stress Resistance and Quality Improvement

[0045] 1. PCR amplification: use the designed primers to amplify the fragment containing the target SNP from the natural population;

[0046] 2. Enzyme digestion: artificially design a HindIII restriction endonuclease recognition site on the primer, and use the restriction endonuclease HindIII to digest the PCR amplification product;

[0047] 3. Separation and identification by agarose electrophoresis: use 4% agarose gel to analyze the digested product and detect its polymorphism. The result is as Figure 4 As shown, this indicates that there is a locus of this SNP in the natural population.

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Abstract

The invention discloses a corn arsenic stress resistant gene ZmAsR2, and an expression product, cloning primer, linkage SNP and traceable SNP cloning primer thereof. The nucleotide sequence of the corn arsenic stress resistant gene ZmAsR2 is shown as SEQID NO:1, and the amino acid sequence of coded protein is shown as SEQID NO:2. The base sequence characteristic of the intramolecular SNP label ofthe corn arsenic stress resistant gene ZmAsR2 is shown as SEQID NO.5. The intramolecular SNP label of the corn arsenic stress resistant gene ZmAsR2 can be used for tracking and identifying the corn arsenic stress resistant gene ZmAsR2, and is suitable for tracking and identification of corn heavy metal stress tolerance field breeding. The corn arsenic stress resistant gene ZmAsR2 is applied to a corn breeding practical method, and can be promoted and applied to identification cloning, and breeding application of other heavy metal stress related genes.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to a corn arsenic stress resistance gene ZmAsR2 and its intramolecular SNP marker and application. Background technique [0002] Due to human activities, such as metal processing, mining, sewage irrigation, and application of herbicides and fertilizers, soil pollution by heavy metals and metalloids has become a worldwide environmental problem. Arsenic is a toxic metal element and has been listed as a Class I carcinogen. Wilson et al. reported that in soil, the concentration of arsenic was less than 10 mg / kg, while in some mining-contaminated soils, the arsenic content was as high as 17400 mg / kg. Since heavy metals in the soil can be absorbed and accumulated by plants and enter the human body through the food chain, high levels of arsenic may pose great risks to human health. Studies by Sharma et al. have shown that the consumption of arsenic-contaminated wheat in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/11C07K14/415C12Q1/6895
CPCC07K14/415C12Q1/6895C12Q2600/156
Inventor 丁冬张雪海李卫华段海洋马拴红王琪月汤继华
Owner HENAN AGRICULTURAL UNIVERSITY
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