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Deletion fragment and application thereof to detection of apple plant dwarfing

A technology of Malus and plants, which is applied in the application field of detecting the dwarfing of Malus plants, and can solve the problems of reduced auxin synthesis, weakened vegetative growth of trees, and insufficient supply of cytokinins in aboveground parts, etc.

Inactive Publication Date: 2017-09-01
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, This leads to weakened tree vegetative growth and reduced auxin synthesis in shoots (li, et al. 2011)

Method used

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  • Deletion fragment and application thereof to detection of apple plant dwarfing
  • Deletion fragment and application thereof to detection of apple plant dwarfing
  • Deletion fragment and application thereof to detection of apple plant dwarfing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1, Obtaining the Deletion Sequence of the IPT5b Promoter of Isopentenyltransferase

[0055] 1. IPT5b promoter sequence analysis

[0056] A search for the isopentenyltransferase IPT5b gene in the apple genome found that there is only one IPT5b gene in the entire apple genome, located on chromosome 16, such as figure 1 shown. figure 1 Show that the basic structure of the gene sequence of the IPT5b protein includes an exon, and the basic structure of the IPT5b sequence includes an exon. For the convenience of describing the gene structure, the IPT5b gene start code in the gene sequence of the IPT5b protein is The position of A in the sub-ATG is recorded as +1, - represents the 5' direction of the IPT5b gene start codon ATG in the genome gene sequence of the IPT5b protein, + represents the IPT5b gene start codon ATG in the genome gene sequence of the IPT5b protein 3' direction.

[0057] 2. Amplification of Apple Rootstock IPT5b Gene Promoter

[0058] The genomic...

Embodiment 2

[0065] Embodiment 2, the application of Deletion fragment in detecting the dwarfing ability of Malus rootstock

[0066] 1. Deletion fragment detection

[0067] 1. Genomic DNA of hybrid offspring with dwarf difference plants obtained

[0068] The hybrid offspring of the F1 generation was obtained by crossing with Begonia as the female parent and M9 as the male parent.

[0069] The leaves of the F1 generation of 15 plants (strain numbers 1-15) with dwarfing difference in the F1 generation hybrid progeny were collected respectively, and the genomic DNA was extracted.

[0070] 2. Deletion fragment detection

[0071] Design and synthesize the following primers: (use the partial sequence on the Deletion fragment as the upstream primer, and then design the downstream primer at about 200bp)

[0072] IPT5b-F: 5'-CCCTACTTAGGGATTCCCATCA-3', (SEQ ID NO: 4)

[0073] IPT5b-R: 5'-TGGGGCAGCTTCTCATTTGC-3'. (Sequence 5)

[0074] Using each genomic DNA obtained in step 1 as a template, and...

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Abstract

The invention discloses a Deletion fragment and its application in detecting the dwarfing of Malus plants. The invention provides a method for detecting the dwarfing ability of a rootstock of a Malus plant, comprising the steps of: detecting whether a deletion fragment is contained in the genome of a Malus plant to be tested, and the dwarfing ability of a rootstock of a Malus plant to be tested containing a Deletion fragment in the genome Higher than the Malus plant to be tested that does not contain a Deletion fragment in the genome; the Deletion fragment is the -1213th position to the -1172th position in the upstream direction of A in the initiation codon ATG of the Malus plant genome IPT5b gene ATG. Experiments of the present invention prove that the presence of the Deletion fragment is negatively correlated with dwarfing traits, and the absence and presence of the fragment can be used to quickly screen dwarf plants in offspring, which is of great significance in the field of plant dwarfism.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a Deletion fragment and its application in detecting the dwarfing of Malus plants. Background technique [0002] Dwarfing and dense planting of apple has the advantages of early fruiting, good quality, convenient management, fast variety update and high economic benefit, and 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, This leads to weakened tree vegetative growth and reduced auxin synthesis in shoots (li, et al. 2011). The gene of isopenteny...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/13
Inventor 韩振海冯轶刘静房鸿成肖遥张新忠王忆吴婷徐雪峰张希
Owner CHINA AGRI UNIV
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