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A kind of SNP molecular marker related to the background color of watermelon peel and its application

A molecular marker and watermelon technology, applied in the biological field, can solve the problems of molecular marker research reports, shorten the breeding process, and few molecular markers, and achieve the effects of improving breeding efficiency, stable amplification effect, and high accuracy

Active Publication Date: 2021-04-20
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

Watermelon is an important horticultural crop. my country is the world's largest producer and consumer of watermelon. However, in the process of watermelon breeding, there are few molecular markers available, and traditional selective breeding is still the main method. The efficiency is low and it is difficult to Quick access to new varieties that meet market demand
The background color of watermelon peel is an important appearance and commodity trait of watermelon. Researchers have studied this trait in the 1930s and 1940s. However, as the most common green-related traits (green and dark green) in the background color of watermelon peel, There is no research report on molecular markers that are closely linked to this trait, so there is a need to carry out research on the background color of watermelon peel and develop molecular markers that can be used in watermelon breeding to shorten the breeding process and improve breeding efficiency

Method used

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  • A kind of SNP molecular marker related to the background color of watermelon peel and its application
  • A kind of SNP molecular marker related to the background color of watermelon peel and its application
  • A kind of SNP molecular marker related to the background color of watermelon peel and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Obtaining of SNP molecular marker sites

[0026] 1.1 Construction of high-density genetic map

[0027] The female parent (9904) with dark green peel background and the male parent (Handel) with green peel background were used to construct a recombinant inbred line population containing 126 individuals through continuous multi-generational selfing. The recombinant inbred line population was sequenced, high-density SNPs were developed using HighMap software, and a high-density genetic map of watermelon was constructed. A total of 178,762 SNP molecular markers suitable for recombinant inbred lines with a depth of not less than 4X were detected between the parents. Through bioinformatics analysis, it was found that the localization interval related to the background color of watermelon peel was located in the region of 2.07Mb at the end of chromosome 8 of the watermelon genome.

[0028] 1.2 Acquisition of SNP sites

[0029] CAPS markers were designed using singl...

Embodiment 2

[0030] Embodiment 2 utilizes this SNP molecular marker to identify the method for the background color of watermelon peel

[0031] 2.1. Extraction of watermelon genomic DNA

[0032] The DNA of watermelon samples to be tested was extracted by conventional CTAB method to remove RNA contamination, and the DNA sample volume was 100 μL. Determination of OD of DNA samples by UV spectrophotometer 260 / 280 Ratio, the value should be between 1.8 and 2.0 to obtain high-quality DNA samples, and the concentration should be diluted to 100ng / μL.

[0033] 2.2. Primer design

[0034] The primers were designed according to the 400bp sequence around the 29880100th base of the No. 8 chromosome of the watermelon genome. For the specific nucleotide sequence, see SEQ ID NO: 1 in the sequence table, and for the primer sequences, see Table 1.

[0035] Table 1 Primer Sequence

[0036]

[0037]

[0038] 2.3 PCR reaction system

[0039] The PCR reaction system is shown in Table 2.

[0040] T...

Embodiment 3

[0049] Example 3 Identification of 105 watermelon germplasm resources using SNP molecular markers

[0050]Using 105 germplasm resources of watermelon with different peel background colors in the National Watermelon Medium-term Bank as materials, the SNP molecular marker of the present invention was used to identify the peel background color. For the specific identification method, refer to the method in the above-mentioned Example 2. The identification results show that the phenotypes of the above 105 watermelon materials are completely consistent with the genotype results, indicating that the SNP molecular marker of the present invention can accurately identify the dark green color of watermelon and the background color of green peel, with an accuracy rate of 100%.

[0051] Among the 105 germplasm resources, there are 31 watermelons with dark green peel background and 74 watermelons with green peel background. The specific varieties and phenotypes are shown in Table 4 below, ...

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Abstract

The invention discloses a SNP molecular marker related to the background color of watermelon peel and its application. The SNP site is located at the 29880100th nucleotide of chromosome 8 of the watermelon genome, and the nucleotide is C or G. The nucleotide polymorphism marker site obtained by the invention can be used to identify the undercolor of the watermelon peel. The SNP molecular marker of the present invention is a SNP molecular marker closely linked with the background color of the watermelon peel developed based on the candidate gene. The SNP molecular marker was used to identify the peel background color of 105 watermelon germplasm resources, and the results showed that the accuracy rate of identification of watermelon with dark green peel background color and green peel background color was 100%. The invention does not need to be identified at the maturity stage of the watermelon fruit, can accurately and quickly identify the background color of the watermelon peel at the seedling stage, can improve the breeding efficiency, shorten the identification time, and is suitable for the genetic identification and breeding of the watermelon.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a SNP molecular marker related to the background color of watermelon peel and an application thereof. [0002] technical background [0003] In traditional selective breeding, because it is difficult to determine the genotype of the offspring, the basis of selection is usually the phenotype of the plant rather than the genotype, the selection time is long, and the phenotype is easily affected by environmental factors, resulting in inaccurate selection and less efficient. Molecular marker-assisted breeding uses molecular markers closely linked to the target shape as a tool to screen target traits, and screens germplasm resources by genotype through molecular markers. It has the advantages of accuracy, speed, and no interference from environmental conditions, avoiding traditional breeding. The blindness of character selection in the process improves the breeding efficiency. [0004] SNP...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 路绪强刘文革李兵兵何楠赵胜杰
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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