A Hansenula spp. strain and its isolation method and application
A technology for yeast strains, cacti, applied in the directions of application, isolation of microorganisms, microorganism-based methods, etc.
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Embodiment 1
[0021] Example 1: Isolation and screening of a strain of Hansenula cactus strain JM-1
[0022] Papaya fruits were collected from papaya planting areas in Hainan as test samples, and 5 g of papaya peels were cut out with a sterile knife, added to 90 mL of LYPD culture medium, shaken at 28 °C and 120 r / min for 2 to 3 h, and plate dilution method was used. Dilute the culture medium to 10 -3 、10 -4 、10 -5 and 10 -6 100 μL of the diluted culture solution was spread on the YPDA solid medium and cultured in a constant temperature incubator at 28°C for 48 hours. Single colonies with different shapes were picked and separated by streaking on the plate until an antagonistic bacteria cake of a single colony was obtained. ℃ refrigerator for later use.
[0023] Taking the pathogenic bacteria of papaya anthracnose cultured for 5 days as the target, the antagonistic bacteria were screened by the plate confrontation method, and the cakes of pathogenic bacteria of papaya anthracnose and th...
Embodiment 2
[0025] Example 2: Identification of a strain of Hansenula spp. JM-1
[0026] 1. First, observe the morphological characteristics of antagonistic bacteria through an optical microscope, such as figure 1 The colony of the antagonistic strain JM-1 of Example 1 on the YPDA medium is milky white and round, with a smooth, moist, shiny surface, neat edges, sticky texture, and an aromatic smell. Observing through a microscope, the cells of the thalli are lemon-shaped or oval, without forming pseudohyphae and spores, and budding at both ends of the thalline.
[0027] 2. Next, refer to the "Characteristics and Identification Manual of Yeast" to carry out physiological and biochemical tests. The antagonistic bacterial strain JM-1JM-1 in Example 1 can ferment glucose, raffinose and fructose; for the determination of carbon source assimilation, JM-1 can assimilate Glucose, sucrose, raffinose, maltose, soluble starch and fructose cannot assimilate L-rhamnose, D-galactose, lactose and citri...
Embodiment 3
[0033] Embodiment 3: Antibacterial test of Hansenula spp. JM-1 on papaya anthracnose pathogenic bacteria
[0034] Select healthy papayas of the same size and disinfect them with 1% (v / v) sodium hypochlorite for 1 min, rinse them with tap water, and dry them in the air. Prick two 1mm deep wounds on the surface of papaya with a punch (10mm in diameter), and divide the fruits into 4 groups: Group A: only inoculate 50 μL sterile water as a control; group B: inoculate 25 μL sterile water and 25 μL concentration of 1 x 10 7 CFU / mL pathogen spore suspension; Group C: inoculate 25 μL sterile water and inoculate 25 μL with a concentration of 1×10 8 CFU / mL yeast suspension; Group D: Inoculate 25 μL with a concentration of 1×10 8 CFU / mL yeast suspension and 25 μL concentration of 1 × 10 7 CFU / mL pathogen spore suspension. The 3 groups of fruits after treatment were placed in a constant temperature and humidity incubator at 25°C and a relative humidity of 90%, and the diameter of lesi...
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