cDNA molecular marker of onion fertility restoration and applications thereof
A technology for molecular markers and fertility recovery, applied in recombinant DNA technology, DNA/RNA fragments, microbial measurement/inspection, etc., can solve the problem of increasing farmers' production burden and planting risks, high production costs of onions, and lack of hybrid generation Variety and other issues, to achieve the effect of saving hybridization test cross time, high cost, and long cycle
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Embodiment 1
[0075] A cDNA molecular marker for restoring onion fertility, the nucleotide sequence of which is shown in SEQ ID NO.1.
[0076] To identify the specific primers for the above cDNA molecular markers, the nucleotide sequence of the upstream primer F240 is shown in SEQ ID NO.2, and the nucleotide sequence of the downstream primer R240 is shown in SEQ ID NO.3.
[0077] The cDNA molecular marker is obtained by the following steps:
[0078] (1) Detection and analysis of total RNA in samples of different tissue materials
[0079] The total RNA of different samples of onion was extracted by Trizol method, and then the total RNA was treated with DNaseI in order to completely remove the residual DNA contamination. The extracted total RNA was detected by spectrophotometer, where OD 260 / OD 280 The ratio is between 1.8-2.0; non-denaturing agarose gel electrophoresis analysis 28s, 18s bands are clear, no diffusion, no DNA residues (such as figure 1 shown). It is confirmed that the ex...
Embodiment 2
[0092] Application of the cDNA molecular markers and specific primers in Example 1 in the identification of onion nuclear interaction male sterility nuclear restoration materials and fertility maintenance materials The above application, the steps are as follows:
[0093] The Trizol method was used to extract the total RNA of the flower buds of the onion sample, and then the total RNA was reverse-transcribed into cDNA, and after PCR amplification with specific primers (upstream primer F240, downstream primer R240), the product was carried out on 1.5% agarose gel. Electrophoresis, voltage 140v, electrophoresis time 30 minutes, after ethidium bromide staining, compare with ethidium bromide staining DNA molecular markers after electrophoresis on 1.5% agarose gel under the same conditions as above, if it is far from the starting point of the swimming lane If there is a band at the same distance, it contains cDNA molecular markers, which is the sample of onion fertility recovery mat...
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