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
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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
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[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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