Problem with Arabidopsis seeds sterilization

Jim Tokuhisa tokuhisa at
Wed Feb 2 06:04:23 EST 2000

At least one important correction needs to be mentioned regarding the
message thread "Problem with Arabidopsis seeds sterilization"

Hygromycin will work only if Arabidopsis is transformed with a
plant-expressed hygromycin phosphotransferase.

>Hi Byung-hoon Kim, I have same experience with you. Something on your
>plate after sterilization might be agrobacteria kept in seed coat.
>Hygromysin work to kill them in my experiment.
>Good luck.

Seed sterilization might be more effective if a 5-20% bleach solution
(0.5-2% sodium hypochlorite) is used rather than potassium chloride.

>Hello everyone,

>I sterilized Arabidopsis seeds which have been transformed by vacuum
>infiltration. But one week after growing on the selection plates I
>a lot of fungi on the plates. I repeated it again and again, but I
>couldn't get rid of them.
>The sterilization methode was :
>       3000 - 4000 seeds per 1.5 ml Epp. tube
>       shaking for 5 min, RT, in (95% EtOH and 0.1% Triton X-100)
>        shaking for 5 min, RT, in (Potassium chloride solution with one
>drop of Tween 20)
>       washing 3 times with sterile H2O
>Could anyone out there give me a tip?
>Thanks in advance for any comment
>Byung-Hoon Kim

Jim Tokuhisa
Max Planck Institut für Chemische Ökologie
Carl-Zeiss-Promenade 10
D-07745 Jena

Phone:  +49-3641-64-36-51
FAX:  +49-3641-64-36-50
E-mail:  tokuhisa at


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