Antagonistic yeast Pichia galeiformis as well as preparation and application method thereof
A technology of Pichia pastoris and strains, which is applied in the field of Pichia pastoris, can solve the problems of lack of antibacterial spectrum strains, and the biocontrol effect is only verified on a few fruits, so as to achieve significant social and ecological benefits and avoid harm to people , easy to cultivate effect
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Embodiment 1
[0017] Example 1: Biological properties of Pichia galeiformis strain BY27
[0018] 1. Morphological features
[0019] (1) YPDA medium (1% yeast extract powder, 2% peptone, 2% glucose, 1.8% agar, sterilized at 121°C for 20 minutes) was cultured at 26°C for 48h, and the colonies were round and white with smooth and round edges. The cell shape is ellipsoidal.
[0020] (2) After culturing in YPDA liquid medium for 24 hours, no mold was formed, the bacterial solution was turbid, and there was precipitation. Microscopically, the yeast cells were oval and budded.
[0021] 2. Molecular biological identification
[0022] Use the universal forward primer NL-1 (5'-GCATATCAATAAGCGGAGGAAAAG-3') and reverse primer NL-4 (5'-GGTCCGTGTTTCAAGACGG-3') to perform PCR amplification of yeast 26S rDNA D1 / D2 region nucleic acid sequence, and the PCR product The sequencing results were entered into the website www.NCBI.nlm.nih.gov for BLAST, the homologous sequences were downloaded from the GenBank...
Embodiment 2
[0024] The inhibitory effect of implementation example 2 Pichia BY27 on apple blue mold, botrytis cinerea and black spot
[0025] 1. Experimental protocol
[0026] Pichia BY27 was taken out from the -80°C refrigerator, activated with YPDA medium (10g of yeast extract powder, 20g of peptone, 20g of glucose, 18g of agar, 1000ml of deionized water, natural pH, sterilized at 121°C for 30min), and picked a single Colony into YPD liquid medium, culture at 26°C and 200r / min for 24h, centrifuge at 4000rpm for 5min, discard the supernatant, wash the collected bacteria repeatedly with sterile water for 3 times, count on a hemocytometer to prepare the concentration 1×10 8 cells / mL of Pichia BY27 suspension.
[0027] Activate Penicillium expansum, Botrytis cinerea or Alternaria tenuissima on PDA medium plate, culture at 26°C for 7-14 days, scrape appropriate amount of spores, wash with sterile water Prepared to a concentration of 5 x 10 4 cells / mL of Penicillium, Botrytis, or Alternar...
Embodiment 3
[0037] The inhibitory effect of implementation example 3 Pichia BY27 on pear fruit blue mold and black spot
[0038] 1. Experimental protocol
[0039] Pichia BY27 was taken out from the -80°C refrigerator, activated with YPDA medium (10g of yeast extract powder, 20g of peptone, 20g of glucose, 18g of agar, 1000ml of deionized water, natural pH, sterilized at 121°C for 30min), and picked a single Colony into YPD liquid medium, culture at 26°C and 200r / min for 24h, centrifuge at 4000rpm for 5min, discard the supernatant, wash the collected bacteria repeatedly with sterile water for 3 times, count on a hemocytometer to prepare the concentration 1×10 8 cells / mL of Pichia BY27 suspension.
[0040] Activate Penicillium expansum or Botrytis cinerea on a PDA medium plate, culture at 26°C for 7-14 days, scrape appropriate amount of spores, and prepare a concentration of 5×10 with sterile water. 4 cells / mL of Penicillium or Botrytis cinerea spore suspension. Healthy and undamaged pe...
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