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Molecular marker related to color property of tomato fruit and application thereof

A molecular marker and tomato technology, applied in the field of genetic engineering and molecular biology, can solve the problems of manpower and material resources, shortage, etc., and achieve the effect of speeding up the breeding process, accurate and reliable results, and simple and effective methods

Active Publication Date: 2014-10-08
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In current breeding, due to the lack of molecular markers related to the genes controlling pericarp color, the screening of different fruit materials often relies on the traditional method of phenotyping at maturity. This method has a strong lag and consumes a lot of manpower and material resources.

Method used

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  • Molecular marker related to color property of tomato fruit and application thereof
  • Molecular marker related to color property of tomato fruit and application thereof
  • Molecular marker related to color property of tomato fruit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Acquisition of SNP-p markers related to tomato fruit color traits

[0049] 360 tomato materials collected from all over the world were resequenced, and each material generated about 5G of data, covering an average of 5 times the tomato genome. 258 individuals with clear phenotypes in the population (232 red fruit and 26 powder Results) Genome-wide association analysis, using a complex linear model (mixture line model), using tassel4.0 software, found a SNP locus highly correlated with peel color phenotype from 1.01 million SNPs ( figure 1 ). The SNP is located at 8616bp upstream of the ATG of the gene Solyc01g079620 (that is, the gene SlMYB12) (SL2.40ch01:71246984). This site C is highly linked to the red fruit phenotype (all 232 red fruits are C), and A is highly linked to the pink fruit phenotype. (25 individuals are A, 1 individual is C).

[0050] Use the primer pair (Seq ID No.2-3) that spans the region containing the SNP-p to detect red fruit and pink f...

Embodiment 2

[0053] Example 2 Obtaining of InDel-p markers related to tomato fruit color traits

[0054] The sequencing data of 20 pink fruits and 20 red fruits were respectively selected from the resequencing materials (the library length was 500 bp during sequencing, and the paired-end sequencing strategy was adopted), and their resequencing data were compared with the tomato reference genome (version SL2) using BWA software. .40) for comparison, and use SAMtools software to convert the comparison result into a readable file. All paired-end reads (paired-end reads) in the region between the gene Solyc01g079620 and its upstream gene Solyc01g079610 (SL2.40ch01:71229871-71,258,882) were detected, and the distance between a pair of reads in all red fruit individuals was found to be within About 500bp, however, there are abnormal reads in the individual pink fruit, and their distance is about 1100bp, indicating that there is a deletion of about 600bp in the individual pink fruit ( figure 2 ...

Embodiment 3

[0059] Example 3 Acquisition of SNP-s and InDel-i markers related to tomato peel color traits

[0060] Perform PCR detection and pyrosequencing on the coding region of the SlMYB12 gene, the specific method is as follows:

[0061] The forward primer used for amplification was F5'-AATAATGGGAAGAACACC-3' and the reverse primer was R5'-TTCTGGACCTAGACTAAA-3'.

[0062] The PCR reaction conditions were: 94°C for 5 minutes; 35 cycles of 94°C for 30 seconds, 55°C for 30 seconds, and 72°C for 30 seconds; 72°C for 10 minutes.

[0063] The amplified product was subjected to agarose gel electrophoresis, stained with ethidium bromide, and observed in a gel imaging system. The target band size was 439bp (Seq ID No.9), and the amplified product was sequenced for detection. .

[0064] Result analysis: the tomato to be tested with base A inserted at 251 in the target band is pink fruit ( Figure 4 ); or the tomato to be tested whose base is T at 311bp is a pink fruit, and the tomato whose bas...

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Abstract

The invention provides a molecular marker related to the color property of tomato fruit and application thereof. The molecular marker includes 2 SNP markers and 2 InDel markers. SNP-p is located at site 8616bp of the upstream of initiator codon of a tomato SlMYB 12 gene, wherein bases which are C represent red tomatoes and bases which are A represent pink tomatoes. SNP-s is located at site 307bp of the downstream of the initiator codon of the tomato SlMYB 12 gene, wherein bases which are T represent pink tomatoes and bases which are C represent red tomatoes. InDel-p is a sequence located at about site 4kp of the upstream of the initiator codon of the tomato SlMYB 12 gene and having a size of 603 bp, wherein pink tomatoes are deleted in the sequence. InDel-i is located at site 248bp of the downstream of the initiator codon of the tomato SlMYB 12 gene, wherein A is located behind the site and is a pink tomato. The molecular marker can be applied to molecular marker-assisted breeding of tomatoes and can substantially accelerate the breeding process of tomatoes.

Description

technical field [0001] The invention relates to the fields of genetic engineering and molecular biology, in particular to molecular markers related to tomato fruit color traits. Background technique [0002] Tomato fruit color is an important agronomic trait that directly affects consumer demand. Early classic genetic experiments proved that the color of tomato fruit is determined by the color of the peel and flesh. The color of the tomato peel is divided into two types: yellow and transparent, which are determined by the y gene. The yellow peel is dominant to the colorless peel. The yellow peel is produced because Yellow flavonoids accumulated in the pericarp during the ripening process, but lacked in the colorless pericarp. The color of the pulp is mainly red and yellow, controlled by the r gene, and the combination of different colored peels and pulps will produce fruits with different appearance colors. The pulp of cultivated tomato is mostly red, because of the differ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 黄三文祝光涛林涛张俊红伦尧尧杜永臣王孝宣
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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