sryknr

Hi everyone,

I have such a big problem unfortunately. I would like to share it and hope that someone will help me.

Whatever i did about sterilization about the suckers, it did not work properly. I mean there are so many contaminated suckers at growth chamber. Last time I tried two stages of sterilization, first the suckers were washed under the sink for 30 mins than i dipped the suckers in %96 ethanol solution for 30 seconds, than %25 NaOCl solution for 15 mins, than rinsed 3 times 1-2-5 mins distiled water. At the end of first sterilization stage the outer layer of suckers are removed. Than at the second stage of sterilization starts and the suckers are waited in %10 NaOCl solution, than tinsed 1-2-5-10 mins distiled water. Than we remove one more layer of banana suckers, than we placed them on the culture media.
But almost all suckers are darkening and bacterial contamination never ends
We used Gentamicin and Na-ampicilin with AgNO3 but it was not enough.

The other problem is multiplication. I mean we use BAP and 2IP but there is no shoot forming, rarely we observe green leaves and elongation but this is also not what we want.

If you have any idea about solution, that would be great.