Primers and method for purity identification of hybrid seeds of Chinese pumpkin 'Guangmi NO.1'
A technology of hybrid seed purity and Chinese pumpkin, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of unestablished hybrid seed purity identification methods, and achieve high commercial application value, Simple operation and low cost effect
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Embodiment 1
[0025] Establishment of a method for the detection of seed purity of hybrid pumpkin 'Guangmi No. 1'.
[0026] 1. Screen the SSR primers for purity identification.
[0027] The published pumpkin EST-SSR primers were screened between the parents (the male parent Changqing 2 and the female parent Zaoxuan 1), and the primer VRISSRPMK8 with a co-dominant differential marker band was selected. The sequence is as follows:
[0028] VRISSRPMK8-F: 5'-TAATTAAGGCGTGAAATTGG-3' (SEQ ID NO: 1)
[0029] VRISSRPMK8-R: 5'-GAAAAGAAACATAAACAGGGC-3' (SEQ ID NO: 2)
[0030] The marking band is clear and repeatable. Primer VRISSRPMK8 can produce 193bp maternal specific marker VRISSRPMK8 193 and the 187bp paternal-specific marker VRISSRPMK8 187 .
[0031] 2. Using the above-mentioned specific primers to identify the purity of hybrid pumpkin 'Guangmi No. 1' seeds.
[0032] (1) Extraction of pumpkin DNA
[0033] The experimental materials are the cotyledon DNA of the commercial variety of pumpki...
Embodiment 2
[0058] Adopt the method of embodiment 1 to 100 strains ' Guangmi No. 1 ' that get from the seed purity identification field of Baiyun base, number its single plant, extract single plant DNA and detect (see figure 2 ), the test results are all true hybrids, the seed purity is 100%, which is consistent with the field investigation results, and the accuracy rate is 100%.
Embodiment 3
[0060] Adopt the method of embodiment 1 to 100 strains ' Guangmi No. 1 ' that get from the seed purity appraisal field of Vegetable Institute base, number its single plant, extract single plant DNA and detect (see image 3 ), the results showed that among 100 samples, there were 97 hybrid seeds and 3 female parents, and the seed purity was 97%, which was consistent with the field investigation results.
[0061] The above examples show that the method of the present invention can effectively distinguish the hybrid seeds of 'Guangmi No. 1' from the seeds of its female parent (Zaoxuan No. 1), and quickly and accurately detect the purity of the seeds.
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