Metarhizium anisopliae|BNCC144648 |BNCC

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Metarhizium anisopliae

Culture medium	Comprehensive PDA agar (CPDA): potato boil 1.0L, glucose 20.0g,KH2PO4 3.0g,MgSO4 · 7H2O 1.5g, vitamin B1 trace, agar 20.0g,pH 6.0±0.2. Sterilization at 121 ℃ for 15min. Potato boiling liquid: weigh 200g of peeled potato pieces, boil in boiling water for 30min, and collect the filtrate to a constant volume of 1.0L.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it in the safety 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.
Growth conditions	28 ℃;7-10 days; Aerobic;
Storage conditions	2-8 ℃
morphology	Small filamentous fungi, with obvious colonies on comprehensive PDA medium, white hyphae at the initial stage, dense and vigorous, and pale yellow and green spores at the later stage. subs, spread to the edge of the plate and grow, the back of the medium is yellow.
Sharing mode	Public welfare sharing

Metarhizium anisopliae

Storage conditions: 2~8 ℃

No. 144648

Product format: freeze dried, 200ul

Validity : 6 years

Biosafety level: 1 , handle in ultra-clean 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 plate 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, integrated PDA,7-10 days. Comprehensive PDA: potato boiling solution 1.0L, glucose 20.0g,KH2PO4 3.0g,MgSO4·7H2O 1.5g, vitamin B1 trace, agar 20.0g,pH 6.0±0.2. Sterilization at 121 ℃ for 15min. Potato boiling solution: weigh 200g of peeled potato pieces, boil in boiling water for 30min, and collect filtrate to a constant volume of 1.0L.

Recovery steps:
(1) Prepare 1-2 of above mentioned plates;
(2) Open it in the biosafety cabinet, sterilize of the ampoule, heat the tip in a flame, quickly drop sterile water to creak it, then break it with forceps;
(3) draw 0.5ml of sterile water into the freeze dried ampoule, make it fully dissolved, and distribute the solution to the plates well in 200ul/plate;
(4) Put the plates under the above culture conditions for cultivation for 7-10 days.
Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability:	good viability, in 7 days strain layer become obvious
colony morphology:	small filamentous fungi have obvious colonies on the comprehensive PDA medium, the initial hyphae are white, dense and vigorous, and then produce light yellow green spores, which spread to the edge of the plate and grow, and the back of the medium is yellow.
Conclusion:	good viability, no abnormal colony morphology, qualified