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An indel marker for apple fruit acidity and its application in molecular-assisted selection

A DNA molecule and apple technology, applied in an InDel marker and in the field of molecular-assisted selection, can solve the problems of restricting apples, complex genetic background, and long childhood, so as to shorten the breeding period and improve the screening efficiency.

Active Publication Date: 2021-06-01
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Due to the characteristics of complex genetic background, long childhood, and self-incompatibility of apple, the genetic breeding of apple and the selection of new varieties are seriously restricted.

Method used

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  • An indel marker for apple fruit acidity and its application in molecular-assisted selection
  • An indel marker for apple fruit acidity and its application in molecular-assisted selection
  • An indel marker for apple fruit acidity and its application in molecular-assisted selection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1. Discovery of Molecular Markers

[0061] The content of malic acid in the mature apple pulp of the mapping population was determined by high performance liquid chromatography (HPLC). Based on the data of the malic acid content and the application of high-density genetic maps for QTL mapping, the acidic trait was located in the 1M interval of chromosome 8. . According to the resequenced SNP markers, the QTL segment was shortened by genetic methods, combined with gene functions, and finally a gene related to apple pulp malic acid content was cloned, namely MdSAUR37 gene.

[0062] Further research found that there is a 36bp insertion polymorphism in the MdSAUR37 gene, which is located upstream of the start codon in the MdSAUR37 gene in the genome, specifically between the -639th and -640th nucleotides (in the MdSAUR37 gene, the start codon The A in the codon ATG is counted as the first position, the adjacent nucleotide upstream of the start codon is the -1st pos...

Embodiment 2

[0066] Example 2. Application of molecular markers to identify malic acid content traits of offspring of high-acid parents and low-acid parents

[0067] 1. Preparation of Hybrid Populations

[0068] 1. Take the pollen of "Golden Delicious" apples, pollinate the emasculated "Red Jade" apples, and harvest hybrid seeds.

[0069] 2. Cultivate hybrid seeds to obtain apple seedlings.

[0070] The planting location was Changping District, Beijing, and 1600 apple seedlings were obtained.

[0071] 2. Determination of malic acid content in fruit

[0072] Mature fruits were collected for identification when the seedlings began to bear fruit.

[0073] 1. After the fruit on the apple plant is ripe, remove the peel and core, and take the pulp.

[0074] 2. Weigh 5g (fresh weight) of the pulp obtained in step 1, add 10ml of double-distilled water, grind into a homogenate, then bathe in 75°C water for 30min, then centrifuge at 12000rpm for 10min, collect the supernatant and leave the preci...

Embodiment 3

[0110] Embodiment 3, application of molecular markers to identify existing varieties

[0111] Carry out following operation respectively to 92 existing apple varieties (natural groups):

[0112] Detect fruit malic acid content, method is the same as step 2 of embodiment 2. The chromatograms of some varieties when detecting the malic acid content of fruits are shown in Figure 5 .

[0113] For genotype identification, the method is the same as Step 3 in Example 2.

[0114] According to the genotype, 92 varieties were divided into three groups, and the average fruit malic acid content of each group was counted. The results are as follows:

[0115] S PI36 S PI36 There are 26 genotype varieties, and the fruit malic acid content is 7.62±1.89mg / g.

[0116] S PI36 S P There are 47 genotype varieties, and the fruit malic acid content is 4.93±1.88mg / g.

[0117] S P S P There are 19 genotype varieties, and the fruit malic acid content is 2.49±0.65mg / g.

[0118] see results ...

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Abstract

The invention discloses an InDel marker for acidity of apple fruit and its application in molecular assisted selection. The present invention firstly protects a molecular marker, which is the following (a1) or (a2): (a1) the DNA molecule shown in sequence 1 of the sequence listing; (a2) the homology between apple and sequence 1 of the sequence listing Greater than 95%, 98%, or 99% of the DNA molecules. The invention also protects the application of molecular markers in predicting the acid content traits of apples. The invention also protects a method for predicting the acid content traits of apples, comprising the following steps: detecting the genotype of the apples to be tested based on molecular markers, S PI36 S PI36 Acid content of genotype apple >S PI36 S P Acid content of genotype apple >S P S P Acid content of genotyped apples. In view of the characteristics of long childhood and complex genetic background of apples, the invention will greatly improve the screening efficiency of apple breeding and shorten the breeding period.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an InDel marker for acidity of apple fruit and its application in molecular assisted selection. Background technique [0002] Apple is one of the most important cultivated tree species in the world because of its rich nutrition and high economic value. At the same time, apples are favored by people because of their crisp, juicy flesh, sweet and sour flavor, and strong aroma. The organic acid in apple fruit is mainly malic acid. Organic acid is one of the important factors determining the quality of fruit flavor. Organic acid participates in photosynthesis, respiration, and synthesis of phenols, amino acids, esters and aromatic substances in the fruit's own metabolism. metabolic process. Therefore, it is of great significance to study the mechanism of organic acid accumulation in apple fruit to improve and improve fruit quality. At the same time, the fruit quality direc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张新忠贾东杰韩振海申飞王忆孙瑞黄振宇吴贝王旋旋吴婷员相焱王吾谦胡慧瑶刘丽陈驰杰张玖利张希许雪峰
Owner CHINA AGRI UNIV