Rhizobium leguminosarum|BNCC225149 |BNCC

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Rhizobium leguminosarum

Culture medium	Rhizobium YMA medium (YMA):KH2PO4 0.25g,K2HPO4 0.25g,MgSO4 · 7 H2O 0.2g,NaCl 0.1g, yeast extract 0.8g, mannitol 10.0g, agar 18.0g, distilled water 1.0 L,pH 7.2. Sterilization at 121 ℃ for 15min.
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	30 ℃,2-3 days, aerobic
Growth characteristics	1. Individual morphological characteristics: rod-shaped, moving, flagella, sandwich, Gram-negative, containing PHB, fission; 2. Culture characteristics: round medium-protruding colonies, translucent, can be cultured in liquid; 3. Physiological and biochemical characteristics: chemical energy organic nutrition, aerobic, no need of light, PH neutral, good growth at 28 degrees Celsius, symbiotic nitrogen fixation.
Storage conditions	2-8 ℃
morphology	Size: 1-2mm Shape: Round Edge: Irregular Transparency: Translucent Color: light white uplift: middle convex surface: smooth and bright texture: moist and viscous
application	Symbiotic nitrogen fixation with white clover, it has been widely used in southern provinces since 1984.
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