Method and specific primer pairs for authenticating Morchella importuna mating types
A kind of technology of Morchella latifolia and specific primer pair, applied in the field of biology, can solve problems such as loss of mating type, and achieve the effect of obvious social benefit
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Embodiment 1
[0032] Example 1 Identification of the mating type of the monoascospore population of Morchella spp. YPL6
[0033] The monoascospore strain YPL6-1—YPL6-24 of Morchella rhizome YPL6 was inoculated on PDA medium, cultured at 23°C for 10 days, picked appropriate amount of mycelia, extracted DNA by CTAB method, and took 3 μL in 1% (W / V) Agarose gel electrophoresis to detect DNA quality and concentration.
[0034] Using the DNA of YPL6-1—YPL6-24 as a template, PCR amplification was performed on P8-4F / P8-4R with primers. The 25 µL amplification system included 2×PCRmix (TSINGKE Bio Inc) 12.5 µL, 10 µM / L 1 µL each of P8-4F and P8-4R primers, 1 µL of DNA template, ddH 2 O 9.5 µL. The PCR amplification program was as follows: pre-denaturation at 94°C for 3 min; denaturation at 94°C for 30 sec, annealing at 60°C for 30 sec, extension at 72°C for 90 sec, 30 cycles; extension at 72°C for 10 min.
[0035] The DNA of YPL6-1—YPL6-24 was used as a template respectively, and the P6-1F / P6-1...
Embodiment 2
[0037] Embodiment 2 The identification of the effectively cultivated strains of Morchella thalamus
[0038] The strains to be tested are Morchella sp. sp1, SP2, YPL6, and YPL7. The strain cultivation, DNA extraction, PCR amplification system and amplification procedure are the same as in Example 1.
[0039] The amplified product was detected by electrophoresis on a 1.2% (W / V) agarose gel, and the results are shown in image 3 , SP2, YPL6, YPL7 strains have bands in PCR amplification with P8-4F / P8-4R, P6-1F / P6-1R primers, indicating that they have two mating types and are effective cultivation strains. The SP1 strain has no band when amplified by the P8-4F / P8-4R primer pair, but has a band amplified by the P6-1F / P6-1R primer pair. It is a MAT1-2 mating type strain and cannot be used as a cultivated strain. Strains YPL6 and YPL7 had fruited after cultivation.
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