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CAPS molecular marker for identifying background color of watermelon peel and application thereof

A molecular marker and peel technology, used in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of being easily affected by the environment and low efficiency, saving manpower and material resources, high sensitivity, and improving efficiency Effect

Active Publication Date: 2019-01-22
ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are few research reports on the background color of watermelon peel, and the selection of watermelon peel background color is mainly carried out through traditional breeding methods for phenotypic identification, which is inefficient and easily affected by the environment

Method used

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  • CAPS molecular marker for identifying background color of watermelon peel and application thereof
  • CAPS molecular marker for identifying background color of watermelon peel and application thereof
  • CAPS molecular marker for identifying background color of watermelon peel and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1, the development of CAPS molecular markers and primers for identifying watermelon peel background color

[0030] Firstly, the fine mapping of skin color-related genes was carried out by constructing a high-density genetic map, as follows: by resequencing 126 individual plants of recombinant inbred lines and their parents (9904 as the female parent and Handel as the male parent), a A high-density genetic map of watermelon. A total of 178,762 SNP markers suitable for RILs with a depth of not less than 4X were detected between the parents. The total length of the map is 1508.94cM, including 11 linkage groups, 2132 Bin markers, and the average map distance is 0.74cM . Through bioinformatics analysis, the candidate genes that control the traits of peel background color were located in the range of 2.07Mb at the end of chromosome 8. By designing CAPS markers and SNP markers for fine mapping, the gene controlling the background color of watermelon peel was locate...

Embodiment 2

[0036] Embodiment 2, utilize above-mentioned CAPS molecular marker to identify the method for the background color of watermelon peel

[0037] 1. Extraction of genomic DNA from watermelon samples

[0038] Use conventional CTAB method to extract genomic DNA from watermelon leaves, remove RNA contamination, use UV spectrophotometer to detect OD values ​​of DNA samples at 260nm and 280nm, and measure DNA concentration and OD 260 / 280 , choose OD 260 / 280 For high-quality DNA samples at 1.8-2.0, the concentration was diluted to 100ng / μL.

[0039] 2. PCR amplification

[0040] The above-mentioned watermelon sample genomic DNA was amplified with the primer set used to amplify the CAPS molecular marker in Example 1; the reaction system for PCR amplification was: 2×PCR Mix 12.5 μL, DNA 2 μL, upstream primer 1.25 μL, downstream Primer 1.25 μL, add deionized water to 25 μL.

[0041] The reaction program of PCR is: 94°C 1min30sec; 94°C 20sec, 57°C 20sec, 72°C 50sec, a total of 30 cycle...

Embodiment 3

[0044] Embodiment 3, the verification of the above-mentioned CAPS molecular markers to the phenotype of the pericarp background color of 105 watermelon germplasm resources

[0045] Taking 105 parts of watermelon germplasm resources collected by Zhengzhou Institute of Fruit Trees, Chinese Academy of Agricultural Sciences as material, using the CAPS molecular markers obtained by the present invention, adopting the method in Example 2 to carry out pericarp background color identification of 105 parts of watermelon germplasm resources, agar Glycogel electrophoresis results are shown in figure 1 . The identification results showed that the phenotypes of 105 watermelon germplasm resources were consistent with the band types of electrophoresis results, and the accuracy was 100%. Therefore, the CAPS molecular marker of the present invention can be used to identify the background color of watermelon peel.

[0046] Among the above-mentioned 105 germplasm resources, there are 31 waterm...

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Abstract

The invention provides a CAPS molecular marker for identifying the background color of watermelon peel. The nucleotide sequence of the CAPS molecular marker in watermelons with dark green background peel color is shown in SEQ ID NO: 1, and the nucleotide sequence of the CAPS molecular marker in watermelons with green background peel color is shown in SEQ ID NO: 2. A primer set for detecting the CAPS molecular marker consists of an upstream primer sequence shown in SEQ ID NO: 3 and a downstream primer sequence shown in SEQ ID NO: 4. The process of identifying the background color of watermelonpeel by using the primer set includes the following steps that genomic DNA is extracted, PCR amplification is carried out, BccI enzyme digestion is carried out, and electrophoresis judgment is carriedout. The CAPS molecular marker has high specificity, the results of identification of the background color of the peel of 105 parts of watermelon germplasm resources show that the accuracy of identification of the molecular marker on the dark green and green background peel colors is 100%, it is indicated that the molecular marker can quickly and accurately identify the background color of the watermelon peel, and the breeding process is accelerated.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a CAPS molecular marker for identifying the background color of watermelon peel and an application thereof. technical background [0002] In the traditional plant breeding process, because it is difficult to obtain the genotype of the offspring of the plant, it is usually only possible to carry out field screening and identification and subsequent cultivation based on the phenotype of the plant. This kind of selection takes a long time, is easily affected by the environment, and depends on the experience of breeders, resulting in long breeding time and deviation from the genotype, low selection efficiency, and poor selection accuracy. Molecular marker-assisted breeding is based on the selection of molecular markers linked to related genes or specific markers of the genes themselves. It has high efficiency, high accuracy, and is not disturbed by environmental factors. It has been proven...

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 李兵兵刘文革路绪强赵胜杰何楠
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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