Radiation-resisting aureobasidium pullulans and application thereof in preparing melanin

[0038] Example 1: Isolation and screening of Aureobasidium pullulans MF1CGMCCNo.CGMCC11275

[0039] The Aureobasidium pullulans used in the present invention are sampled and separated from 5cm-20cm soil on the surface of a certain radiation-contaminated environment in Xinjiang. Take 1.0g of the collected soil samples and put them in 15mm×150mm sterile test tubes, place 60 10kGry was irradiated under Coγ-rays, then 5mL of liquid Chapeburger medium was added to the test tube, and enriched by shaking at 28°C for 3 days. Take 1mL of the above-mentioned enrichment solution and transfer it into a test tube filled with 9mL sterile physiological water. After successively performing gradient dilution, take 0.2mL of the diluted solution and spread it on a Chase culture medium plate, and cultivate it at a constant temperature of 28°C for 7 days, and observe the colonies For growth status, a single colony was picked for purification. Some of the purified strains were stored in lyophiliz...

[0043] Example 2: Isolation, screening and identification of Aureobasidium pullulans MF1CGMCCNo.CGMCC11275

[0044] 1. PCR amplification of the rDNAITS segment of Trichoderma viride and its sequencing

[0045] The strains were inoculated in modified Chapeauer's medium, cultured at a constant temperature of 28°C for 4 days, and the bacteria were collected by centrifugation. The total DNA was extracted according to the extraction method of Gerrits et al., and the fungal general primers in the D1 / D2 region of LSUrDNA were used for PCR amplification. The primers were designed as follows:

[0046] NL1: 5′-GCATATCGGTAAGCGGAGGAAAAG-3′,

[0047] NL4: 5'-GGTCCGTGT-TTCAAGACGG-3';

[0048] The PCR products were purified and recovered by gel cutting, dissolved in deionized water, and sequenced.

[0049] 2. Sequence alignment and phylogenetic analysis of the D1 / D2 region of LSUrDNA

[0050] The LSUrDNA D1 / D2 region sequence obtained from the strain was compared with the known sequence i...

[0051] Example 3: Utilize Aureobasidium pullulans (Aureobasidium pullulans) MF1CGMCCNo.CGMCC11275 to extract and prepare crude melanin extract

[0052] (1) Activation of strains: Aureobasidium pullulans MF1CGMCCNo.CGMCC11275 was inoculated on the improved modified Chapei solid medium, and cultured at 28° C. for 48 hours to activate the strains.

[0053] Described improved Cha's solid medium: sodium nitrate 3g, yeast extract powder 0.5g, dipotassium hydrogen phosphate 1g, magnesium sulfate (MgSO 4 ·7H 2 O) 0.5g, potassium chloride 0.5g, ferrous sulfate 0.01g, glucose 30g, agar 20g, distilled water 1000mL, pH natural, subpackage, and sterilize.

[0054] (2) Cultivation of strains: inoculate the activated bacterial strain in step (1) into the improved Chapei liquid medium, wherein the loading amount of the improved Chapei liquid medium is 100mL, glucose 10.3g / L, and yeast extract powder 2.02g / L, ammonium sulfate 2.0g / L, NaCl 0.5g / L, inoculum size 2mL, 30°C, 180r / min shaker cul...