Agrobacterium rhizogenes K599

K599發根農桿菌菌株

Description

Agrobacterium rhizogenes strain K599 (pRi2659), a causative agent of hairy root disease, effectively induces

hairy root 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. Obtain an NUTRIENT AGAR plate with the appropriate antibiotic.

2. Using a sterile pipette tip, touch the bacteria growing within the punctured area of

the stab culture. (A sterilized wire loop or sterile toothpick can be used in place of a

sterile pipette tip.)

3. Run this tip lightly over a section of the plate, as shown in the figure, to create streak

#1.

4. Using another sterile pipette tip, pass through streak #1 and spread the bacteria over a second section of the

plate, to create streak #2.

5. Using a third sterile pipette tip, pass through streak #2 and spread the bacteria over the last section of the plate,

to create streak #3.

6. Grow overnight in a 28 o C incubator (unless a different growth temperature is indicated on the plasmid

datasheet).

7. In the second morning, single colonies should be visible. If the bacterial growth is too dense, re-streak onto a

new agar plate to obtain single colonies.