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.
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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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