New strain of Grifola frondosa
A technology of Grifola frondosa and new strains, applied in the direction of fungi, microorganisms, microorganisms, etc., can solve the problems of extinct production, undifferentiated primordia, poor adaptability, etc.
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Embodiment 1
[0052] The specific steps for obtaining the hybrid strain are as follows:
[0053] (1) Collection of basidiospores and acquisition of monokaryotic strains
[0054] 1) Pick the fruiting body caps of the parental strains Qinghui 151 and Tihui No. 1 that are seven mature (the cap is fan-shaped and forms a tube hole),
[0055] Put it in a ziplock bag and write a label.
[0056] 2) Place the cap collected in step 1) in a sterilized petri dish on the ultra-clean operating table, cut off the stipe, divide it into several pieces of appropriate size, and set aside.
[0057] 3) Take the sterilized PDA medium, pour it into a sterilized culture dish and cover it with a little when it is about to solidify, take the Grifola frondosa mushroom cap prepared in step 2), bury the upper surface in the medium, wait for the medium to solidify, and fix it The fruiting body cap, with the bacterial tube facing down, is buckled on the lower half of the petri dish, sealed with Parafilm, and at the bot...
Embodiment 2
[0069] The hybrid strain obtained in Example 1 is screened, and the specific steps are as follows:
[0070] 1) Preparation of liquid strains
[0071] Add 300mL of liquid culture medium to a 500mL Erlenmeyer flask, and activate the hybrid strains and parental strains obtained in the above step (3) that were activated on a PDA-enriched medium plate (activation conditions were cultured at 25°C in the dark for 10 days). For Hui 151 and Tihui No. 1, use a puncher with a diameter of 5mm to make the same quality and amount of bacteria blocks, inoculate them into the liquid medium, the inoculation amount is 8 bacteria blocks / bottle, seal the air-permeable sealing film, and keep at 25°C 160r·min -1 Cultivate for 14 days to obtain liquid strains.
[0072] 2) Cultivation method
[0073] The compost is mixed by a mixer and then put into polypropylene bags (18cm×35cm) with an automatic bagging machine. Each bag contains 1.2kg of wet material, 19cm in height, and a ring at the mouth of t...
Embodiment 3
[0087] Using ISSR and SRAP molecular markers to identify the new strain 86 (T5Q9) of Grifola frondosa, the specific steps are as follows.
[0088] 1) Mycelia Genomic DNA Extraction
[0089] Inoculate the preserved Grifola frondosa strain in PDA-enriched medium for activation for 10 days, insert it on a plate culture medium covered with sterilized cellophane, and culture it in the dark at 25°C for 10 days. Gently scrape off the hyphae with tweezers and place in 1.5 In mL centrifuge tubes, weighed and set aside; UNIQ-10 column type fungal genomic DNA extraction kit was used to extract mycelia genomic DNA, and the purity and concentration of DNA were detected by agarose gel electrophoresis and biospectrophotometer.
[0090] 2) Primers
[0091] The 10 ISSRs and 7 pairs of SRAP primers used are shown in Table 3 and Table 4:
[0092] The primer combinations of SRAP are: me1 / em2, me3 / em1, me3 / em3, me3 / em6, me5 / em1, me5 / em3, me5 / em5.
[0093] Table 3 Primer sequences for ISSR marke...
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