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Specific sequence and its molecular markers and identification method of tomato fruit dry juice traits

A technology of tomato and dried juice, applied in the field of genetics and biology, to improve the efficiency of selection and speed up the breeding process

Active Publication Date: 2020-04-17
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

However, there are few reports on the use of molecular marker technology to directly identify whether tomatoes are dried juice tomatoes in the prior art.

Method used

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  • Specific sequence and its molecular markers and identification method of tomato fruit dry juice traits
  • Specific sequence and its molecular markers and identification method of tomato fruit dry juice traits
  • Specific sequence and its molecular markers and identification method of tomato fruit dry juice traits

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Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1: The specific sequence of a tomato fruit dry juice trait provided by the present invention, the sequence is shown in SEQ ID NO: 1, the molecular marker for identifying the trait, including an upstream primer, a downstream primer, and a core of the upstream primer The nucleotide sequence is shown in SEQ ID NO:2, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO:3.

[0026] The present invention obtains a batch of SNP sites and indel sequences linked with dried juice traits by resequencing the dried juice tomato genome, designs primers for these sites and sequences, conducts genetic linkage analysis and verification, and screens out a series of Molecular markers linked to dry juice traits, including CAPS, dCAPS and PCR markers. Since dried juice tomato is a recessive trait, co-dominant markers were selected for verification, including CAPS-1, CAPS-2 and dCAPS-1.

[0027] CAPS-1:

[0028] Forward primer: 5'-TAGGCAAACTCTATTCATCAAGG-3'...

Embodiment 2

[0045] Embodiment 2: In addition to disclosing the specific sequence and primers of the above-mentioned dried tomato fruit juice traits, the present invention also discloses a method for identifying the dried tomato fruit juice traits, including the following steps:

[0046] (1) using the genomic DNA of the tomato to be identified as a template, carrying out PCR amplification with the molecular marker to obtain its amplified product;

[0047] (2) Carry out agarose gel electrophoresis detection to the amplified product, if the PCR amplified product of the tomato to be identified contains the DNA fragment shown in SEQ ID NO: 1, then it can be identified that the tomato to be identified is a juicy tomato , otherwise it is dried juice tomato.

[0048] The PCR reaction temperature program in the step (1) is: pre-denaturation at 94°C for 4 minutes; denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 45 s, 30 cycles; extension at 72°C for 8 min.

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Abstract

The present invention discloses a tomato fruit all-flesh trait specific sequence, which is represented by SEQ ID NO:1. The invention further provides a method for identifying the tomato fruit all-flesh trait, and a molecular marker, wherein the molecular marker comprises an upstream primer represented by SEQ ID NO:2 and a downstream primer represented by SEQ ID NO:3. The method comprises: (1) carrying out PCR amplification with the molecular marker by using the genomic DNA of tomato to be identified as a template; and (2) carrying out agarose gel electrophoresis detection on the amplification product, wherein the identifying result shows that the tomato to be identified is the juice tomato if the PCR amplification product of the tomato to be identified contains the DNA fragment represented by SEQ ID NO:1, and otherwise, the identifying result shows that the tomato to be identified is the all-flesh tomato. According to the present invention, with the application of the molecular marker and the identification method to perform the molecular marker-assisted selection of the tomato fruit all-flesh trait, the selection efficiency can be improved, and the breeding process can be accelerated; and the sample can be identified only through simple DNA extraction, PCR specific amplification and agarose gel electrophoresis detection.

Description

technical field [0001] The invention belongs to the technical field of genetics and biology, and in particular relates to a tomato fruit dried juice trait-specific sequence, a molecular marker and an identification method thereof. Background technique [0002] Tomato has high nutritional value and economic value. It is an important fresh food product and processing raw material. It has become the seventh largest crop in the world after corn, rice, wheat and potato. The global annual industrial output value can reach 50 billion US dollars. Tomato fruit not only contains carbohydrates, cellulose and mineral elements that are beneficial to the human body, but also contains a variety of vitamins, as well as a variety of sugars, organic acids and volatile organic compounds that determine the flavor of the fruit. Because of its important agricultural value, rich nutritional value, superior botanical characteristics and basic research advantages, tomato has become a model plant for...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 李君明刘磊舒金帅王佳彤白金瑞冯晶晶
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI