Abnormal pichia anomala and application thereof
A Pichia pastoris, abnormal technology, applied in the field of microorganisms
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Embodiment 1
[0061] Isolation and strain identification of Pichia anomala YPL6 of the present invention
[0062] 1. Sample collection
[0063] Fruits are collected from organic apple orchards in China.
[0064] 2. Isolation and screening of strains and antagonistic screening
[0065] Bacteria were isolated from organic fruits, separated by conventional gradient dilution coating, and cultured on PDA (containing streptomycin sulfate) and YPD agar (containing streptomycin sulfate) at 28°C, respectively, and the colonies were picked. Bacteria with large morphological differences and pure culture were purified and stored on YPD agar medium, and the primary screening and repeated screening of antagonistic bacteria were carried out with Botrytis cinerea as the target pathogen, and a strain with strong antibacterial activity was finally obtained. Yeast strain, named YPL6.
[0066] 3. Identification of strains
[0067] The morphological, physiological and biochemical characteristics and some co...
Embodiment 2
[0072] Determination of the antibacterial spectrum of Pichia anomala YPL6 of the present invention
[0073] Tested pathogens:
[0074] Peach brown rot fungus--Monilinia fructicola (wint.) Rehm;
[0075] Botrytis cinerea (Botrytis cinerea Per.ex Fr) after apple harvest;
[0076] Apple postharvest Penicillium--Penicillium expansum Link, 1809;
[0077] Pear postharvest Penicillium--Penicillium expansum Link, 1809;
[0078] The above pathogenic bacteria were selected as indicator fungi, and the bioassay method was used for artificial perforation in the fruit body (taking the control of apple postharvest botrytis as an example to illustrate).
[0079] The specific operations are as follows:
[0080] The artificial perforation inoculation method in the fruit is as follows: the pathogenic bacteria strains are incubated in YPD liquid medium at 28°C for 48h, and the concentration of the bacteria liquid is adjusted to 1×10 with distilled water. 8 cfu / ml. B. cinerea was cultured on...
Embodiment 3
[0086] Determination of the inhibitory effect of Pichia anomala YPL6 on diseases of the present invention
[0087] The effect of YPL6 on postharvest disease control of apples, peaches and pears under storage conditions.
[0088] 1. After picking, the apples are soaked in YPL6 bacterial solution for 30-60 seconds, then air-dried at room temperature, and then stored in the dark at a temperature of 1-2°C and a relative humidity of 90-95%. After 10 months of storage, the control effect was calculated compared with the control group. When the cell concentration is ≥10 8 When cfu / ml, the control effect is more than 80%; when the cell concentration is 10 7 When cfu / ml, the control effect is 60%; when the cell concentration is 10 6 When cfu / ml, the control effect is 50%. The effective concentration is ≥10 7 cfu / ml.
[0089] 2. After picking, the peaches are soaked in YPL6 bacterial solution for 30-60 seconds, then air-dried at room temperature, and then stored in the dark at a te...
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