Primer and method for identifying purity of cucumber 'Yuexiu No.3' hybrid seed
A technology for hybrid seed purity and cucumber, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., can solve problems such as false hybrids, decreased seed genetic purity, production loss, etc., to achieve low cost, High commercial application value, simple operation effect
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Embodiment 1
[0032] Example 1 Establishment of a method for testing the purity of hybrid cucumber 'Yuexiu 3' seeds.
[0033] 1. Screen the SSR primers for purity identification.
[0034] Screen between the parents of the published cucumber SSR primers, and select two sets of primers for co-dominant differential marker bands, wherein,
[0035] The sequence of primer set 1 is as follows:
[0036] 3B-10-F: 5'-CATCACGACCCTCTCCATCT-3' (SEQ ID NO: 1);
[0037] 3B-10-R: 5'-GGGTTTGATAGTGGAGATTATTCA-3' (SEQ ID NO: 2);
[0038] The sequence of primer set 2 is:
[0039] 2B-41-F: 5'-CACCGAGAACGAAAGAAGGA-3' (SEQ ID NO: 3);
[0040] 2B-41-R: 5'-TCTAAGGTGGGAGGGAAACC-3' (SEQ ID NO: 4).
[0041] The marker bands of the two sets of primers are clear and reproducible, and the primers can produce maternal-specific markers and paternal-specific markers.
[0042] 2. Using the above-mentioned specific primers to identify the purity of hybrid cucumber 'Yuexiu 3' seeds.
[0043] (1) Extraction of Cucumber...
Embodiment 2
[0068] Embodiment 2 specificity experiment
[0069] Take the seeds of the same type of cucumber varieties on the market: Zhongnong 108, Yuanfeng, Jinlv 5, Zhongnong 8, Jinyan 4, Yuexiu 3, female parent A-2, male parent C-8, and extract them respectively DNA was amplified by PCR with primer set 1 and primer set 2, respectively. The results showed that only Yuexiu 3 had two specific bands, and other varieties had only one band or no band. image 3 is the electropherogram of the amplification product of primer set 1, Figure 4 Electropherogram of the amplification product of primer set 2.
Embodiment 3
[0070] Example 3 field application
[0071] Using the method of Example 1, 42 strains of 'Yuexiu No. 3' taken from the seed purity identification field of Baiyun base were numbered, and the DNA of the individual strains was extracted for detection. The results showed that the detection results of the two sets of primers were consistent, and the seed purity was 97.6%, which was consistent with the field investigation results, and the accuracy rate was 100%. Figure 5 is the electropherogram of the amplification product of primer set 1, Figure 6 Electropherogram of the amplification product of primer set 2.
[0072] The above examples show that the method of the present invention can effectively distinguish the 'Yuexiu No. 3' hybrid seeds from their parent seeds or seeds of other cucumber varieties, and quickly and accurately detect the purity of the seeds. And choose any one of the primers can be accurately identified.
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