SNP locus and primer combination for identifying germplasm authenticity of cucumis melo, and application

A technology of authenticity and location, applied in applications, plant products, recombinant DNA technology, etc., can solve problems such as infeasible methods, time-consuming and laborious, and long identification cycles

Active Publication Date: 2020-12-04
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional field planting identification cycle is long, easily affected by environmental factors, time-consuming and laborious
Therefore, in the face of the melon germplasm resources with large quantity and serious homogeneity, the method of field identification is not feasible

Method used

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  • SNP locus and primer combination for identifying germplasm authenticity of cucumis melo, and application
  • SNP locus and primer combination for identifying germplasm authenticity of cucumis melo, and application
  • SNP locus and primer combination for identifying germplasm authenticity of cucumis melo, and application

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0079] The preparation method of the above kit includes packaging each primer in any one of the above primer sets separately.

[0080] In a fourth aspect, the present invention provides the aforementioned DNA fingerprint database of muskmelon germplasm resources based on core SNP markers, wherein the DNA fingerprint database includes: genotypes of core SNP sites of standard muskmelon germplasm resources.

[0081] The above-mentioned standard melon germplasm resources include 441 melon germplasm resources selected from the following:

[0082]PLANTERE JUMLED 76134, PERSIAN MEDIUM, CUNTELOWPE PLANTERE JUMBO, 76W-2-5, 76W3-3, CANTA LOLLPE, 76W5-1, 76W4-2, 76W3-4, CNTALOU PE P.M.R450, CASABA GOLDEN BEAUT, Zhangzhou Melon , サカェやまぶき*ロニ, サカタプリンスメロPF, サカタアンテスメロン, サカタェェ-クンメロン, Japan1, gold mating Askamelon, Japan2, POLIOOR F (RS 815), CANTOR HYBRIDE F1, Itada PEpi- タ-タソロン, Itada Nie-Meron, Itada ピ-スメロン, Nagaoka ス-パ-Meron, Hane-デュ, CA4 VR 6072, CA2 VGK 456, CA9M761232, (MUSKMELON) BURPEE...

Embodiment 1

[0100] Acquisition of loci and primer combinations for identifying the authenticity of melon germplasm resources

[0101] 1. Discovery of 32 core SNP sites

[0102] Based on the resequencing data of 1175 melon germplasm resources, the present invention screens out 150 core germplasm resources through population structure and principal component analysis. Using these 150 core germplasm resources, 32 SNP sites were obtained through deep mining. These 150 melon germplasm resources include two subspecies and 6 varieties. Including the main types of germplasm resources in the Chinese market, there are huge differences in many traits such as fruit shape, flesh color, and peel color, and the genetic diversity is high.

[0103]The screening criteria for core SNP sites are as follows: (1) SNP sites with MAF0.3 in the resequencing SNP data set were removed to obtain 1,124,208 SNPs. (2) In the result of (1), remove the SNP sites with other mutations in each 30 bp of the left and right...

Embodiment 2

[0122] This example is the validity test of the SNP primer combination developed in Example 1, and it is also the construction of the DNA fingerprint database of melon germplasm resources based on the above-mentioned 32 core SNP markers.

[0123] The 441 muskmelon germplasm resources tested in this example are all common excellent germplasm resources or germplasm resources imported from abroad, and are all stored in the germplasm resource bank of the Vegetable Center of Beijing Academy of Agriculture and Forestry Sciences. The details are shown in Table 3 below.

[0124] Table 3: Planting Resource Sources

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135] 1. Obtaining the genomic DNA of the tested muskmelon germplasm resources:

[0136] The leaves of 441 muskmelon germplasm resources for testing were extracted by the cetyltrimethylammonium bromide (CTAB) method (30 seeds were taken from each germpla...

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Abstract

The invention belongs to the field of molecular markers and detection thereof, and particularly relates to a core SNP locus and primer combination for identifying the germplasm resource authenticity of cucumis melo, an SNP locus-based DNA fingerprint database and database construction method, and an SNP locus-based cucumis melo germplasm resource authenticity identification method and application,wherein SNP loci are selected from any one to 32 of the first SNP locus to the thirty-two SNP locus. A DNA fingerprint database for identifying the germplasm resource authenticity of the cucumis melobased on high-throughput sequencing is established in the invention. The method provided by the invention not only can be used for identifying unknown cucumis melo germplasm resources, but also can be used for identifying the authenticity of known cucumis melo germplasm resources. The method provided by the invention has the advantages of high flux, accuracy, low cost, simplicity in operation, manpower and material resource saving and the like, and has a very wide application prospect.

Description

technical field [0001] The invention belongs to the field of molecular markers and detection thereof, in particular to a core SNP site for identifying the authenticity of melon germplasm resources, a combination of primers, a DNA fingerprint database based on the SNP site, a database construction method, and a melon based on the SNP site Identification methods and applications of the authenticity of germplasm resources. Background technique [0002] Germplasm resources are the foundation of the seed industry. Article 92 of the "Seed Law" states that germplasm resources refer to the basic materials for the selection and breeding of new plant varieties, including cultivated species of various plants, propagation materials of wild species, and artificially created materials of various plants using the above-mentioned propagation materials. genetic material. Germplasm resources are an important natural resource formed by natural evolution and artificial creation. They have acc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156A01H5/08
Inventor 温常龙杨静静张建赵泓张晓飞王超罗江
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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