Agrobacteriumrhizogenes K599, 發(fā)根農(nóng)桿菌K599菌株

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Description

Agrobacterium rhizogenes strain K599(pRi2659), a causative agent of hairy root disease, effectively induces hairyroot formation in a variety of plant species, including numerous soybean(Glycine max) cultivars.

Growth Temperature: 25°C;

Medium: NUTRIENT AGAR

Peptone   5.0     g

Beef extract      3.0     g

Agar          15.0            g

Distilled water           1.0     L

Adjust pH to 7.0.

Rehydration fluid: NUTRIENT BROTH

Peptone   5.0     g

Beef extract      3.0     g

Distilled water           1.0     L

Adjust pH to 7.0.

Recovery

1.      Obtainan NUTRIENT AGAR plate with the appropriate antibiotic.

2.      Usinga sterile pipette tip, touch the bacteria growing within the punctured area ofthe stab culture. (A sterilized wire loop or sterile toothpick can be used inplace of a sterile pipette tip.)

3.      Runthis tip lightly over a section of the plate, as shown in the figure, to createstreak #1.

4.      Usinganother sterile pipette tip, pass through streak #1 and spread the bacteriaover a second section of the plate, to create streak #2.

5.      Usinga third sterile pipette tip, pass through streak #2 and spread the bacteriaover the last section of the plate, to create streak #3.

6.      Growovernight in a 28 o C incubator (unless a different growth temperature is indicatedon the plasmid datasheet).

7.      Inthe second morning, single colonies should be visible. If the bacterial growthis too dense, re-streak onto a new agar plate to obtain single colonies.