Identification method and identification kit for different species of Populus species
A kind of kinship and identification method technology, which is applied in the field of identification methods and identification kits for different kinship relationships among Populus sects, can solve the problems of poor stability, complex identification methods, etc., and achieves the effect of rapid method and easy automation.
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Embodiment 1
[0038] S1, using the 5 pairs of polymorphic microsatellite DNA primer combinations (as shown in Table 1) of the present invention to determine the genotype individuals of Populus tomentosa (taken from the National Populus tomentosa germplasm resource bank) of 25 common species of the five poplar schools in my country Perform genotyping and construct microsatellite DNA fingerprints of interspecies individuals, such as figure 2 shown. After the fingerprint is made, it can be used for follow-up experiments without having to make it again every time.
[0039] in, figure 1 Shown is the use of Multiplex Manager software to design primer combinations based on fragment length; figure 2 The fragment lengths of the amplified products are shown to construct the microsatellite DNA fingerprints of different species of Populus genus, and the sizes of the amplified fragments of the five pairs of primers in the primer set correspond in sequence from low to high figure 1 Populus SSR Primer...
Embodiment 2
[0048] Using microsatellite DNA molecular marker technology to identify the male parent of the hybrid progeny of Populus silver glandularis.
[0049] Three adult female plants of Populus silver gland were selected from the poplar mixed forest, and the capsules were collected, and all 10 adult male plants near the female plants were collected at the same time. The seeds in the capsules were propagated by tissue culture. After 3 months, the tissue cultured seedlings were transplanted into nutrition cups, hardened in the artificial climate chamber, and then transferred to the greenhouse seedbed for unified maintenance. Collect 2-3 pieces of young leaves from a single plant.
[0050] The DNA of all parents was extracted by CTAB method, and 20 progeny were selected from each female plant to extract DNA, totaling 73 plants. The DNA was diluted to 10 ng / μl as a template for multiplex PCR.
[0051] All DNA samples were amplified using the previously described microsatellite DNA mark...
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