Trifolium rhizobium|225149 |BNCC

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Rhizobium leguminosarum

Subculture procedure	① Prepare 1-2 of the above plates; ② After disinfecting the surface of the ampoule tube, open it in a safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Take 0.5mL of sterile water and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into a Agar plate at a rate of 200 μ L per piece and apply evenly; ④ Place the agar plate under the above cultivation conditions for cultivation.
Growth conditions	30 ℃; 48-72h; aerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 1-2mm, circular, with irregular edges, semi transparent, light pink, protruding in the middle, smooth and bright surface, sticky and moist texture, easy to pick up, G - (red), Bacillus, purity: pure
Sharing mode	Public welfare sharing

Rhizobium leguminosarum

Storage conditions: 2~8 ℃

BNCC No.： 225149

Product format： freeze dried, 200ul

Validity period: freeze-dried tube 24 months

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:28-30 ℃, aerobic, rhizobia YMA medium. Rhizobium YMA medium: KH₂PO₄ 0.25g,K₂HPO₄ 0.25g,MgSO₄·7H₂O 0.2 g,NaCl 0.1g, yeast extract 0.8g, mannitol 10.0g, agar 18.0g, distilled water 1.0 L,pH 7.2,121 ℃,15min.

Recovery steps:

(1)Prepare 1-2 pieces of YMA plates;

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

(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)Put the agar plates under the above culture conditions for cultivation 4-7days

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 YMA plate colony is obvious
colony morphology:	size: general color: white shape: round edge: neat edge wet and dry: wet, sticky and smooth: smooth transparency: translucent uplift: bulge
conclusion:	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified

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