SNP (single nucleotide polymorphism) molecular marker related to watermelon peel background color and application of SNP molecular marker

A molecular marker and molecular marker-assisted technology, which is applied in application, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of few molecular markers, low breeding efficiency, difficulty in obtaining breakthrough new varieties, etc., and achieve accelerated breeding Process, high accuracy, and the effect of improving accuracy

Active Publication Date: 2018-06-22
ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Watermelon is an important horticultural crop. my country is the world's largest producer of watermelon. However, at present, my country's watermelon breeding is still dominated by traditional breeding. The application of molecular marker-assisted selection is less, and the breeding efficiency is low. It is difficult to obtain breakthrough new varieties.
Watermelon peel traits are an important commodity trait, and they are also the focus of improvement by breeders. As early as the 1930s and 19

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  • SNP (single nucleotide polymorphism) molecular marker related to watermelon peel background color and application of SNP molecular marker
  • SNP (single nucleotide polymorphism) molecular marker related to watermelon peel background color and application of SNP molecular marker

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Example 1 Obtaining of SNP sites closely linked to watermelon peel background color traits

[0019] 1.1. BSA of hybrid population (mixed pool resequencing)

[0020] Using a yellow-peeled watermelon (94E1) as the female parent and a green-peeled watermelon (Qingfeng) as the male parent for cross-breeding, an F2 segregating population containing watermelon with yellow rind background and watermelon with green rind background was prepared, and the population was separated from F2 generation. Select 30 watermelons each with a yellow peel background and a green peel background, extract the leaf genomic DNA, and mix the genomic DNA of 30 watermelons with yellow peel background and 30 watermelons with green peel background. The DNA is mixed together. It was sequenced with the Illumina HiSeq 2000 sequencer to obtain 33.75G data, which was compared with the watermelon reference genome 97103 (http: / / cucurbitgenomics.org / organism / 1). 247012 SNP loci were identified, and the SNP-index...

Embodiment 2

[0027] Example 2 Using SNP molecular markers to identify watermelon peel background color

[0028] 2.1, DNA extraction

[0029] Use conventional CTAB method to extract genomic DNA from watermelon leaves and remove RNA. The volume of genomic DNA sample is 50μL. Measure the OD value of genomic DNA sample at 260nm and 280nm with ultraviolet spectrophotometer, and calculate DNA content and OD 260 / 280 Ratio, select OD 260 / 280 Genomic DNA samples with values ​​between 1.8 and 2.0, diluted to 100ng / μL.

[0030] 2.2, primer

[0031] The primers are the primer pair of SNP molecular marker (SNP02) obtained in Example 1 closely linked to the background color traits of watermelon peel. The primer pair is designed based on the upstream and downstream sequence of the 48456th nucleotide of chromosome 4 of the watermelon genome. The primer sequences are shown in Table 2. The primers were synthesized by Shenggong Bioengineering (Shanghai) Co., Ltd. and diluted to 10μM for use.

[0032] Table 2 Primer ...

Embodiment 3

[0045] Example 3 Using SNP molecular markers to identify the peel background color of the hybrid population

[0046] 3.1. Selection of experimental materials

[0047] The 618 individual plants of the F2 population of the cross combination 94E1 (yellow) × Qingfeng (green) were used as experimental materials, of which 94E1 (yellow) was the female parent and Qingfeng (green) was the male parent. The phenotypic test results showed that the F2 population Among them, there are 458 individual plants with yellow skin background and 160 individual plants with green skin background.

[0048] 3.2. Use SNP molecular markers to identify F2 populations

[0049] The SNP molecular marker (SNP02) related to the peel background color on the watermelon 4 obtained in Example 1 was used to identify 618 F2 individual plants. The specific identification method refers to the method in Example 2. The identification result shows that the identification result of the SNP molecular marker is consistent with the...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to an SNP (single nucleotide polymorphism) molecular marker related to watermelon peel background color and an application of the SNP molecular marker. An SNP site is located on the 48456th nucleotide of chromosome 4 of watermelon genome. The SNP molecular marker can be used for identifying the watermelon peel background color and has higher accuracy rate. The SNP molecular marker is utilized for identifying the watermelon peel background color of a hybrid F2 population, and a result indicates that the identification accuracy rate of the SNP molecular marker for the watermelon peel background color of hybrid population generations is 100%. The SNP molecular marker is utilized for identifying the peel background color of 20 parts of watermelon germplasm resources, and a result indicates that the identification accuracy rates of the SNP molecular marker for watermelons with yellow peel background color andwatermelons with green peel background color are both 100%. Therefore, the SNP molecular marker can rapidly and accurately identify the watermelon peel background color and can be applied to production on a large scale.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to SNP molecular markers related to the background color of watermelon peels and applications thereof. Background technique [0002] In the process of traditional plant breeding, people usually only rely on phenotypes for field selection and subsequent breeding, not only relying on the experience of breeders, but also often affected by the interaction of genotype and environment. Since it is difficult to know the genotype of the offspring, this selection often takes a long time and may deviate from the genotype. It is only suitable for selection of simple quality traits. However, for quantitative traits, the lack of a clear correspondence between phenotype and genotype can easily cause inaccuracy and low efficiency in selection. Molecular marker-assisted selection is an auxiliary means of applying molecular markers to selection in the process of crop variety improvement. It mainl...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11A01H1/04
CPCA01H1/04C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 赵胜杰刘文革豆峻岭路绪强何楠
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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