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MdARF3 gene promoter deletion fragment and application thereof in malus plant dwarf detection

A technology of apple genus and promoter, applied in the direction of DNA / RNA fragments, application, plant gene improvement, etc., can solve the problems of weakened vegetative growth of trees, decreased auxin synthesis, insufficient supply of cytokinins in aerial parts, etc.

Active Publication Date: 2020-04-28
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The dwarfing mechanism of apple dwarf rootstock may be the result of mutual regulation of cytokinin and auxin between the root system and the shoot, that is, the reduction of cytokinin synthesis in the root system of the dwarf rootstock, resulting in insufficient supply of cytokinin in the shoot, As a result, the vegetative growth of the tree body is weakened, and the synthesis of auxin in the shoot is reduced.

Method used

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  • MdARF3 gene promoter deletion fragment and application thereof in malus plant dwarf detection
  • MdARF3 gene promoter deletion fragment and application thereof in malus plant dwarf detection
  • MdARF3 gene promoter deletion fragment and application thereof in malus plant dwarf detection

Examples

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Effect test

Embodiment 1

[0074] Example 1. Obtaining the Deletion Sequence of the Auxin Response Factor Gene MdARF3 Promoter

[0075] 1. MdARF3 promoter sequence analysis

[0076] Searching for the auxin response factor gene MdARF3 gene in the apple genome, it was found that there is only one MdARF3 gene in the whole apple genome, which is located on chromosome 5, such as figure 1 shown. figure 1 Show, the basic structure of the gene sequence of MdARF3 protein comprises 10 exons and 9 introns, in order to describe gene structure conveniently, in the gene sequence of MdARF3 protein, in the MdARF3 gene start codon ATG The position of A is recorded as +1, - represents the 5' direction of the MdARF3 gene start codon ATG in the genome gene sequence of the MdARF3 protein, and + represents the 3' direction of the MdARF3 gene start codon ATG in the genome gene sequence of the MdARF3 protein.

[0077] 2. Amplification of Apple Rootstock MdARF3 Gene Promoter

[0078] The genomic DNAs of dwarf apple rootstock...

Embodiment 2

[0085] Example 2, Application of MdARF3 Gene Promoter Deletion Fragment in Detection of Malus Rootstock Dwarfing Ability

[0086] 1. Detection of MdARF3 gene promoter deletion fragment

[0087] 1. Genomic DNA of hybrid offspring with dwarf difference plants obtained

[0088] The hybrid offspring of the F1 generation was obtained by crossing with Begonia as the female parent and M9 as the male parent.

[0089] The leaves of the F1 generation of 10 plants (strain numbers 1-10) with dwarfing difference in the F1 generation hybrid progeny were collected respectively, and the genomic DNA was extracted.

[0090] 2. Detection of MdARF3 gene promoter deletion fragment

[0091] Design and synthesize the following primers: (Use the upstream sequence of the MdARF3 gene promoter deletion fragment as the upstream primer, and then design the downstream primer at about 500bp)

[0092] MdARF3-F: 5'-GACGATATCAATTAGATTGATGATGTC-3' (SEQ ID No. 4);

[0093] MdARF3-R: 5'-CATGTCGTGGTTGTGTCTCT-3...

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Abstract

The invention discloses an MdARF3 gene promoter deletion fragment and application thereof in malus plant dwarf detection. The invention provides a method for detecting the stock dwarfing ability of amalus plant. The method comprises the following steps of: detecting whether the genome of a to-be-detected malus plant contains the MdARF3 gene promoter deletion fragment or not, wherein the stock dwarfing ability of the to-be-detected malus plant of which the genome does not contain the MdARF3 gene promoter deletion fragment is higher than the stock dwarfing ability of the to-be-detected malus plant of which the genome contains the MdARF3 gene promoter deletion fragment, and the MdARF3 gene promoter deletion fragment is from -1061st to -992nd site in an upstream direction of A in the initiation codon ATG of the malus plant genome MdARF3 gene. A deletion phenomenon of the MdARF3 gene promoter deletion fragment is positively related to dwarf characters, and a dwarf plant in offspring can bequickly screened by use of the deletion and the existence of the fragment so as to perform an important meaning in a plant dwarfing property field.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a missing fragment of an MdARF3 gene promoter and its application in detecting the dwarfing of Malus plants. Background technique [0002] Due to the advantages of early fruiting, good quality, convenient management, fast variety update and high economic benefits, apple dwarfing and dense planting cultivation has become the trend of modern apple production and development. So far, the utilization of dwarfing rootstocks is still the main way to realize the dwarfing and dense planting of apples. [0003] The dwarfing mechanism of apple dwarf rootstock may be the result of mutual regulation of cytokinin and auxin between the root system and the shoot, that is, the reduction of cytokinin synthesis in the root system of the dwarf rootstock, resulting in insufficient supply of cytokinin in the shoot, As a result, the vegetative growth of the tree body is weakened, and the synthesis of auxin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11A01H1/04
CPCC12Q1/6895A01H1/04C12Q2600/13C12Q2600/156
Inventor 韩振海冯轶王忆吕佳红高敏许雪峰张新忠吴婷
Owner CHINA AGRI UNIV
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