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Application of IRT1 promoter insertion element in apple

A promoter, apple technology, applied in the biological field, can solve the problem of unclear internal reasons of differential expression

Active Publication Date: 2016-08-10
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The expression of MxIRT1 gene is different at different times of iron deficiency, but the underlying reason for the differential expression is still unclear

Method used

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  • Application of IRT1 promoter insertion element in apple
  • Application of IRT1 promoter insertion element in apple
  • Application of IRT1 promoter insertion element in apple

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1, Obtaining the deletion element of the iron transporter IRT1 promoter

[0047] 1. Sequence analysis of IRT1 promoter and coding region

[0048] IRT1 was searched in the apple genome, and it was found that there were two IRT1 genes with a CDS similarity of 100% in the entire apple genome, and they were both located on chromosome 5 with a distance of 36735bp, such as figure 1 shown. figure 1 showed that the basic structure of the genome gene sequence of the IRT1 protein includes three exons ( figure 1 The dark solid box in ) and two introns ( figure 1 The black line connecting the dark solid box), in order to facilitate the description of the gene structure, the position of A in the IRT1 gene start codon ATG in the genomic gene sequence of the IRT1 protein is recorded as +1 in the following examples, and - represents IRT1 The 5' direction of the start codon ATG of the IRT1 gene in the genome gene sequence of the protein, + represents the 3' direction of the s...

Embodiment 2

[0060] Example 2, capillary electrophoresis analysis of the separation of TATA-box in apple hybrid progeny

[0061] 1. Crossbreed with Xiaojinhaitang as the female parent and Shandingzi as the male parent, cross with Xiaojinhaitang as the female parent and M9 as the male parent, and the F1 hybrid offspring of Xiaojinhaitang×M9 and Xiaojinhaitang×Shandingzi have etiolation For different plants, the leaves of these plants were collected separately, and the genomic DNA was extracted.

[0062] 2. Design and synthesize the following primers:

[0063] IRT1-F: 5'-AAGCCACCGGAGGACATGG-3', (SEQ ID No.3)

[0064] IRT1-R: 5'-GGGCTATGATTTTGAGGGGCA-3'. (SEQ ID No.4)

[0065] Add FAM fluorescent label to the 5' end of IRT1-F, and finally obtain fluorescent-labeled IRT1-F.

[0066] 3. Using each genomic DNA obtained in step 1 as a template and fluorescently labeled IRT1-F and IRT1-R as primers, perform PCR amplification to obtain each PCR amplification product.

[0067] PCR amplification...

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Abstract

The invention discloses an application of an IRT1 promoter insertion element in an apple. The invention discloses a method for identification or auxiliary identification of the iron-deficiency-resistant degree of a plant. The method comprises detecting a genome gene sequence of an IRT1 protein of a to-be-detected plant. The iron-deficiency-resistant performance of a to-be-detected plant having the genome gene sequence, the promoter area of which has a 5'-ATTATAA-3' sequence, of the IRT1 protein is larger or selectively larger than that of a to-be-detected plant having the genome gene sequence, the promoter area of which doesn't have a 5'-ATTATAA-3' sequence, of the IRT1 protein. According to deficiency and existence of an element TATA-box of the IRT1 promoter, descendant iron-deficiency-resistant plants can be quickly screened. The application makes a significant sense in the field of the iron-deficiency-resistant performance of a plant.

Description

technical field [0001] The invention relates to the application of an IRT1 promoter insertion element in apples, and belongs to the field of biotechnology. Background technique [0002] Iron is one of the essential trace elements for crop growth. Leaf yellowing and premature senescence of fruit trees caused by iron deficiency stress can cause crop quality decline and yield reduction, which has a great impact on agricultural economic benefits. Most of the apple trees in my country grow in alkaline soils, which are prone to yellow leaf disease. Therefore, it is more and more important to seek the internal mechanism of iron deficiency stress to improve the iron deficiency chlorosis of apples. [0003] The divalent cation transporter gene MxIRT1 gene (Peng li, et al, 2006) cloned from the cDNA library of the root system of apple iron-efficient genotype Crabapple iron deficiency treatment (Peng li, et al, 2006). plays an important role in. The full-length cDNA of the gene is 1...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68
Inventor 吴婷张美玲韩振海张新忠王忆许雪峰
Owner CHINA AGRI UNIV
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