Rhizobium vitis|BNCC134955 |BNCC

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Rhizobium vitis

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 ℃,18-24h, aerobic
Storage conditions	2-8 ℃
morphology	Size: 1-2mm Shape: Round Edge: Irregular Transparency: Translucent Color: off-white Protuberance: Middle Protuberance Surface: Bright and Smooth Texture: Wet and Easy to Stir
application	Type strain
Sharing mode	Public welfare sharing

Rhizobium vitis

Storage conditions: 2~8 ℃

No. 134955

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: 30℃, aerobic, 24h, soybean rhizobia culture medium: mannitol 10-20 g, potassium dihydrogen phosphate (KH2PO4) 0.5 g, sodium chloride (NaCl) 0.2 g, magnesium sulfate (MgSO4 7H2O) 0.2 g, Calcium sulfate (CaSO4·2H2O) 0.1 g, calcium carbonate (CaCO3) 1.0 g, yeast liquid (10:100) 100 ml, agar 18-20 g, clear tap water 900 ml. 121℃, 15min.

Recovery steps:

(1)Prepare 1-2 pieces 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 for18-24 hours.

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

Item	test results
viability	good viability，in 20 hours,liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology:	Size: 1-2mm Shape: Round Edge: Irregular Transparency: Translucent Color: off-white uplift: middle convex surface: bright and smooth texture: moist and easy to stir
Conclusion:	good viability, no abnormal colony morphology, qualified

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