Botrytis cinerea|119830 |BNCC

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Botrytis cinerea Pers.

Subculture procedure	① Dissolve the frozen tube in a 30 ℃ water bath and shake quickly for dissolution; ② Wipe the outer wall of the Frozen vial with an alcohol swab for disinfection, and then transfer it to a safe cabinet for operation; ③ Unscrew the tube cap, aspirate all the dissolved solution and its bacterial blocks, and transfer them into 1-2 agar plates (approximately 200 μ L per Agar plate); ④ After uniform coating, transfer to the above cultivation conditions for cultivation.
Growth conditions	28 ℃; 5-7 days; aerobic
Storage conditions	-80 ℃
Safety level	1
morphology	Small filamentous fungi have obvious colonies on comprehensive PDA medium, producing white hyphae in the early stage and light brown in the later stage, with a light brown color on the back of the medium
Sharing mode	Public welfare sharing

Botrytis cinerea Pers

Storage conditions: 2~8 ℃

No. 119830

Product format： freeze dried, 200ul

Validity period: 6 years

Biosafety level: 1, handle in ultra-clear table or safety cabinet

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the bacteria are well. The freeze dried culture shall be used up once and shall not be retained. The bacterial viability will resume after 1-2 generation of recovery and can be used normally. Handling instructions of agar slant and suspension liquid is enclosed. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.

Growth conditions: 28 ℃, aerobic, 5-7 days, comprehensive PDA agar medium: potato boiling liquid 1.0L, glucose 20.0g, KH2PO4 3.0g, MgSO4 7H2O 1.5g, vitamin B1 trace, agar 20.0g, pH 6.0±0.2. Sterilize at 121°C for 15 minutes. Potato cooking liquid: Weigh 200g peeled potato pieces, boil in boiling water for 30min, collect the filtrate and dilute to 1.0L.

Recovery steps:

(1)Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates;

(2)Open the ampoule in the biosafety cabinet, heat the tip of ampoule in a flame, quickly drop sterile water to creak it, then break it with forceps;

(3)Draw 0.5mL of liquid culture medium into a freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly;

(4)Inoculate a plate with 0.2mL of suspension liquid, repeat the step to obtain two plates;

(5)Put all the liquid test tubes and plates under the above culture conditions for cultivation, and the strains can be used when they grow.

Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability	good viability, in 5 days strain layer become obvious, colony is typical
colony morphology:	filamentous fungi, with obvious colonies on comprehensive PDA medium, and white hyphae, Dense tall and long, hyphae are vigorously spreading and growing, and a large number of gray-black spores are produced
Conclusion:	good viability, no abnormal colony morphology, qualified

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