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by the ethanol

Dear Justine Toulotte, 

You should try another approach for disinfection besides GUS and ethanol. Yours plant is infected with oomycetes and it may perpetrate in plant cells and may reproduced both by sexually and asexually.

Hope the like may help you:

http://link.springer.com/article/10.1007/s10327-010-0252-3

https://www.ncbi.nlm.nih.gov/pubmed/9432703

wish you good luck  

Dear Anita Kumari,

Sorry if my question was not clear :)

I'm working in controlled lab conditions and I'm studying plant-oomycete interactions. I don't want to desinfect my samples, I just want to stop the infection (the growth of the oomycete) at a certain time.

I have a gene candidate and I want to see the localisation of its induction upon infection. Therefore I inoculated the plants and after 16 hours of infection, I stained them with a GUS solution to reveal where is it induced (I'm using the promoter of he gene candidate driving the GUS gene). Let's say that I keep the plants during 6 hours in the GUS solution, and then I wash and conserve my samples in EtOH 100%. Should I consider that my infection time is indeed 16hours or 16+6=22hours? (Is the infection stopped by the Gus solution or by the ethanol washing bath?).

Thanks!

Justine

Hi Justine,

The best way is probably to extract RNA and qPCR your gene of interest in addition to the pro:GUS. That said, you could fix your sample in para/gluta before the GUS staining (a little bit of optimization required). Good luck ! PM

Hi Pierre-Marc,

The qPCR has been done :) I just need to see the localization of the expression now. I will have a look at your fixation proposition.

Thanks

Justine