Contamination rate high! - tissue culture (Dec/14/2006 )
Dear all,
I'm currently working on the tissue culture of an alkaloid-producing plant. However, contamination rate was pretty high. I've tried using fungicide and 20% chlorox for sterilizing my leaf explants but to no avail.
Is there any method to control and reduce contamination rate? the contaminants observed were all fungi. I've tried leaving my media for one week prior to culture. No media contamination was observed before i use them for culturing.
Furthermore, i managed to get callus, however, the callus jus turned browned and died after few weeks. Is there any possible reasons for such observations?
Thank you for your attention.
Anyone with solutions can email me: eiggammaggie87@hotmail.com
Mameemoo
I'm currently working on the tissue culture of an alkaloid-producing plant. However, contamination rate was pretty high. I've tried using fungicide and 20% chlorox for sterilizing my leaf explants but to no avail.
do you put any surfactant (like silwet) during the surface sterilization? Chlorox 20% is really arsh for the plant tissue. the thing is that your sterilization procedure should vary slightly depending on the tissue and the plant. Can you sterilize the seed (using chlorine gas for example) and germinate the plant asceptically?
Is there any method to control and reduce contamination rate? the contaminants observed were all fungi. I've tried leaving my media for one week prior to culture. No media contamination was observed before i use them for culturing.
Furthermore, i managed to get callus, however, the callus jus turned browned and died after few weeks. Is there any possible reasons for such observations?
What media were you using? callus induction media and callus maintenance media might be different for your plant.
Thank you for your attention.
Anyone with solutions can email me: eiggammaggie87@hotmail.com
Mameemoo
[HI
are u using in vitro germinated explant or from field? why dont u try invitro germinated plant parts for ur work? that might give u satisfactory results.
u can contact me at shankarmadhan@yahoo.co.uk if u need any more help.
regards
Madhan
Professor n Biotech
PSGCAS
India
In response to Dr T. :
Thank you for your reply!
I've actually added 6 drops of Tween 20 in 20% chlorox. The explants used in this experiment are leaves. The callus induction media i prepared was MS basal medium supplemented with various concentrations of PGRs like TDZ, 2,4-D and Kin. However, callus induced was pretty low and they actually turned brown after few weeks of culturing.
Is there any problems with my method mentioned above?
You can email me at eiggammaggie87@hotmail.com
Once again, thank you for your attention to my problems.
Yours sincerely,
mameemoo
are u using in vitro germinated explant or from field? why dont u try invitro germinated plant parts for ur work? that might give u satisfactory results.
u can contact me at shankarmadhan@yahoo.co.uk if u need any more help.
regards
Madhan
Professor n Biotech
PSGCAS
India
Thank you for your reply! I've sent you an email. thank you!