I am going to apply drought stress on the cell suspension culture, using by PEG8000. So, I will add different concentrations of the PEG into the media and then plant the cells in it to see the response. May I add PEG as powder in the prepared media and autoclave it like what we usually do for a solid medium with agar? Of course I don't need that concentrated media. let say I want to make 10 or 15% PEG. I have found no literature talking about the way they use PEG in suspension culture.
Any suggestion?
best
Dear Mahbobeh zamani babgohari,
I too do impose stress by PEG 6000 in such a way in liquid Yoshida Nutrient medium solution for rice research, but for my work i am not autoclaving the media,just preparing the media n use it . But adjusting the pH regularly is must! Add PEG directly to the media n thoroughly mix before use. I am getting excellent results for the pilot study.
All the best.
Try to prepare liquide media (PEG in powder + liquid) and then measure electroconductivity with deionized water as a control.
Dear colleague,
If you need very sterile media you should autoclave it. I think that for cell culture is needed. To resolve this problem check the osmotic potential before and after autoclaving. If it is the same osmotic potential it means that nothing changes during aqutoclaving.
All the best
Dear Babgohari,
I am very much convinced by the suggestion made by Swapan, but I will suggest to filter sterilized your PEG solution, using a 0.45 mm filter, before using it for suspension culture, PEG autoclave is not recommended . Avoid adding PEG solution directly to the suspension cell. Make a PEG solution of higher concentration and adjust to final concentration needed by diluting it with the autoclaved sterilized media followed by adding suspension cell to it. If possible use PEG 6000, which has been extensively used for drought stress plant. I am attaching one research article where, the relationship with osmotic potential, temp and PEG concentration has been established. Hope it to be helpful for u.
Dear Mahbobeh,
I am going to attempt something similar. I have a question, did you use full strength MS liquid media or did you use half strength??
Thanks.
I would suggest not to autoclave it. Autoclave your media and while it is warm, around 90 degrees add the PEG. This is what I do and I never had problems with contamination.
If you want to prepare solid media with PEG, that is a problem, cause once you add the PEG, the agar will not solidify anymore. so you have to preapre the plates and then pour the PEG, leave it overnight and discard. this of course has nagtive sides, for example you have to spend time figuring out how much of PEG to add to obtain required water potential.
Good luck!!!
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