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Functional molecular marker of a related gene Aft for synthesis of solanum lycopersicum anthocyanin and application thereof

An anthocyanin and genotype technology, which can be used in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., and can solve problems such as co-separation of Aft sites.

Active Publication Date: 2019-05-14
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently some molecular markers linked to Aft, and most of these markers are designed based on the nucleotide sequence polymorphisms of the ANT1 gene or AN2 gene, and cannot be 100% co-segregated with the Aft locus

Method used

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  • Functional molecular marker of a related gene Aft for synthesis of solanum lycopersicum anthocyanin and application thereof
  • Functional molecular marker of a related gene Aft for synthesis of solanum lycopersicum anthocyanin and application thereof
  • Functional molecular marker of a related gene Aft for synthesis of solanum lycopersicum anthocyanin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Example 1, Cloning of Tomato Aft Gene and Discovery of Allelic Variation Form thereof

[0104] 1. Analysis of the structure of the R2R3-MYB gene cluster linked to the Aft gene

[0105] Existing studies have shown that the Aft gene is linked to a R2R3-MYB gene cluster on chromosome 10. According to tomato genome data SL2.50 version released by SGN database (URL: http: / / solgenomics.net / ), the R2R3-MYB gene cluster linked to Aft gene on tomato chromosome 10 was analyzed.

[0106] Analysis results see figure 1 . The results showed that there were four R2R3-MYB genes in the R2R3-MYB gene cluster, which were Solyc10g086250 gene, Solyc10g086260 gene, Solyc10g086270 gene and Solyc10g086290 gene. The Solyc10g086250 gene is the reported AN2 gene (Meng X, Wang J-R, Wang G-D, Liang X-Q, LiX-D, Meng QW. (2015) An R2R3-MYB gene, LeAN2, positively regulated the thermo-tolerance in transgenic tomato. J Plant Physiol 175:1-8.), Solyc10g086260 gene is the reported ANT1 gene (Mathews ...

Embodiment 2

[0125] Example 2, the development of molecular markers and polymorphism detection

[0126]1. Development of Molecular Markers

[0127] Molecular markers were developed based on the two allelic variant forms of the Aft gene.

[0128] The nucleotide sequence of the amplified molecular marker is as follows:

[0129] Primer F: 5'-GGTCACTTATTGCTGGGAGA-3' (sequence 3 in the sequence listing);

[0130] Primer R: 5'-CTCCATGTTGCATGGTTGTTG-3' (SEQ ID NO: 4 in the Sequence Listing).

[0131] 2. Polymorphism detection

[0132] A. Polymorphism detection one

[0133] The tomatoes to be tested were Indigo Rose, LA1996 or Alisa Craig.

[0134] 1. Use the Plant DNA Rapid Extraction Kit to extract the genomic DNA of the tomato to be tested.

[0135] 2. Using the genomic DNA in step 1 as a template, perform PCR amplification with the primer pair consisting of primer F and primer R in step 1 to obtain a PCR amplification product.

[0136] PCR reaction system (20 μL): 10 μL of Premix Taq DN...

Embodiment 3

[0151] Example 3, the application of molecular markers

[0152] 1. Application 1

[0153] The tomatoes to be tested were Indigo Rose, LA1996 or Alisa Craig.

[0154] 1. Carry out genotyping of the tomato to be tested according to the method of polymorphism detection one in Example 2.

[0155] 2. Plant the tomato to be tested in the field, manage it in the field regularly, and observe the phenotype of the fruit at the green ripe stage after maturity, and detect the content of total anthocyanins in the peel of the tomato to be tested.

[0156] The results showed that the pericarp of Indigo Rose had a large amount of anthocyanin accumulation, which was purple-black, and the total anthocyanin content was 7.05mg / g fresh weight; the pericarp of LA1996 had a small amount of anthocyanin accumulation, which was purple spots, and the total anthocyanin The content of Alisa Craig is 0.12mg / g fresh weight; the pericarp of Alisa Craig has no anthocyanin accumulation and is green, and the ...

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Abstract

The invention discloses a functional molecular marker of a related gene Aft for synthesis of solanum lycopersicum anthocyanin and application thereof. The provided specific primer pair is composed oftwo primers for amplifying specific DNA segments; the specific DNA segments are provided with target sequences of the primer pair composed of the primer F and the primer R in a solanum lycopersicum genome; the primer F is a single-stranded DNA molecule shown in a sequence 3 of a sequence table, and the primer R is a single-stranded DNA molecule shown in a sequence 4 of the sequence table. By the adoption of the molecular marker, solanum lycopersicum materials containing anthocyanin can be rapidly screened out, and thus the breeding pace of new solanum lycopersicum varieties is accelerated. Thefunctional molecular marker has high theoretical significance and economic value.

Description

technical field [0001] The invention belongs to the field of plant molecular breeding, and in particular relates to a functional molecular marker of tomato anthocyanin synthesis-related gene Aft and its application. Background technique [0002] Anthocyanins and lycopene are two important classes of plant pigments and are well-recognized antioxidants with health-promoting functions. Pathological studies have shown that these two types of substances have the functions of promoting health and preventing diseases, and have obvious effects in the prevention and control of metabolic diseases such as cancer, cardiovascular diseases, and diabetes. [0003] Tomato (Solanum lycopersicum) is an important fruit vegetable crop, which is deeply loved by consumers in various countries. Commonly cultivated tomato fruits generally only accumulate lycopene, not anthocyanins. However, some wild tomato fruits only accumulate anthocyanins, not lycopene. In order to further improve the antiox...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
Inventor 李传友孙传龙邓磊蒋红玲李常保
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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