SCAR marker and method for early identification of tomato internode length

A tomato and internode technology, applied in the fields of biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of difficult new varieties, low efficiency, and few molecular markers, achieving high accuracy and speeding up selection. The effect of low-cost growth and detection

Active Publication Date: 2021-06-04
NORTHEAST AGRICULTURAL UNIVERSITY
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Problems solved by technology

Tomato belongs to Solanaceae, tomato genus, and has become one of the world's vegetable crops. However, there are few molecular markers available in the process of tomato breeding, and traditional selective breeding is still the main method. The efficiency is low, and it is difficult to quickly obtain new ones that meet market demand. Variety

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  • SCAR marker and method for early identification of tomato internode length
  • SCAR marker and method for early identification of tomato internode length
  • SCAR marker and method for early identification of tomato internode length

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Embodiment 1

[0017] Obtaining of embodiment 1 SCAR mark

[0018] Using wild-type tomato 16615 and long-node field natural mutant 16578 as materials, construct P 1 ,P 2 , F 1 and F 2 Four-generation population, using parental resequencing combined with F 2 The group separation method (Bulked Segregant Analysis, BSA) further screened candidate intervals. The specific operation is as follows: select the parent parent P 1 ,P 2 One each, F 2 25 strains in each extreme mixed pool were used for DNA extraction; library construction and sequencing were carried out, and the whole genome resequencing on the BGISEQ platform was completed by Shenzhen Huada Company; further biological information analysis was performed, and the filtered reads were compared with the reference genome. NCBI: https: / / www.ncbi.nlm.nih.gov / genome / gdv / browser / genome / ? id=GCF_000188115.4 as a reference genome. Through bioinformatics analysis, unqualified data is filtered out to obtain high-quality valid data. Finally,...

Embodiment 2

[0026] Application of embodiment 2THH-FOR0 mark

[0027] F 1 , F 1 F 2 group. Using the SCAR marker primer set for 210 copies of F 2 Segregate populations for internode length identification. The specific method is as follows: first, the CTAB method is used to extract leaf DNA of tomato with identification; the leaf DNA is used as a template, and the primer pair in the embodiment is used for amplification. The reaction system for PCR amplification is: 10 μL 2×Mix Master, 1 μL DNA template, 1 μL each of upstream and downstream primers, 7 μL ddH 2O. The reaction program of PCR amplification is: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s; annealing at 56°C for 40 s; extension at 72°C for 50 s; cycle 35 times; extension at 72°C for 10 min; . The amplified products are then identified.

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Abstract

The invention belongs to the technical field of plant molecular markers, and particularly relates to a SCAR marker and method for early identification of a tomato internode length. The SCAR marker TTHH-F0R0 for early identification of the tomato internode length has nucleotide sequences shown as SEQ ID No. 1 and SEQ ID No. 2. The invention further provides an amplification primer pair of the SCAR marker and the method for early identification of the tomato internode length. The SCAR marker closely linked with the tomato internode length is obtained through identification. When the marker is adopted for identification, a plant type condition of the tomato internode length can be accurately and rapidly identified in a seedling stage rather than a stage in which a tomato plant type is stable, and a selective breeding process of tomato varieties with excellent plant types can be greatly accelerated. According to a SCAR marker primer, enzyme digestion is not needed, and only PCR one-step detection is needed, so that the advantages of low detection cost, high accuracy and the like are achieved, and the SCAR marker can be well used as the molecular marker for assisting in selective breeding of tomato internode plant types.

Description

technical field [0001] The invention belongs to the technical field of plant molecular markers, in particular to a SCAR marker for early identification of tomato internode length and an identification method. Background technique [0002] A broad molecular marker refers to an heritable and detectable DNA sequence or protein. Molecular markers in a narrow sense refer to specific DNA fragments that can reflect certain differences in the genomes of organisms or populations. SCAR markers are usually molecular markers transformed from RAPD, SRAP, and SSR markers. Because it does not require enzyme digestion, saves money and time, has good stability and reproducibility, it has become the first choice for molecular marker-assisted selection breeding. Tomato belongs to Solanaceae, tomato genus, and has become one of the world's vegetable crops. However, there are few molecular markers available in the process of tomato breeding, and traditional selective breeding is still the main...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 杨欢欢姜景彬王特许向阳张萍李景富赵婷婷张贺
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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