SSR molecular markers for identifying Paeonia sect. Moutan variety and application thereof
A peony and molecular marker technology, which is applied to the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of being easily affected by the environment, has not yet been reported, and the identification efficiency is low, and achieves reliable and accurate results. , good sensitivity, obvious difference effect
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Embodiment 1
[0046] Example 1 Screening of SSR molecular markers and primer combinations
[0047] First, the present invention selects all 72 pairs of EST-SSRs from the five linkage groups of the first high-density genetic map of peony (Cai, 2015; Wu Jing, 2016), and synthesizes corresponding primers. The genomic DNA of Paeonia japonica was used as template DNA to amplify with these 72 pairs of EST-SSR primers, and the amplified product was detected by 6% denatured polyacrylamide gel electrophoresis combined with silver staining method, and finally screened out polymorphism rich The 34 EST-SSRs, 34 EST-SSRs and their primer information are shown in Table 2.
[0048] Table 2 34 EST-SSR molecular markers and corresponding primer information
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[0052] F = forward primer; R = reverse primer.
[0053] Taking the 264 varieties of P. peony in the prior art as test materials, further screening among the above-mentioned 34 EST-SSRs can realize the minimum number...
Embodiment 2
[0073] Example 2 Using the optimal combination of SSR molecular markers to establish the molecular identity cards of 264 varieties of purple spot peony
[0074] This example provides the optimal combination of 12 SSR molecular markers and their corresponding primer combinations screened in Example 1 to establish molecular ID cards for 264 varieties of Paeonia purpurea.
[0075] In the 12 pairs of primers PS095, PS356, PS166, PS119, PS271, PS068, PS367, PS074, PS157, PS221, PS159, and PS187 obtained in Example 1, all the band patterns of each pair of primers in 264 Paeonia purpurea varieties are divided by Arranged in order from small to large (the size of the amplified fragments of the two alleles is equal and the smallest is the minimum value, the size of one allele fragment remains the same, and the size of the other allele fragment increases in turn, followed by , and then rearrange the fragments that were determined to be constant allele fragments with increased sizes, and...
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