Does anyone have an idea if maize protoplasts could be stored? If yes, do you know the conditions and the efficiency of the protoplast after storage?
Hi,
I asked myself the same question, since I'm doing incorporation experiments and I have to find the optimal conditions to have maximal photosynthetic activity. I therefore decided to measure oxygen production in different storing conditions.
From my experience:
Fresh protoplasts at RT: approx 100 umolO2 /mgChl * h
Fresh protoplasts on ice: approx 100 umolO2 /mgChl * h but rapidly decreasing to 20 after 20 min
After 24 h at...
...RT: completely broken and no oxygen evolution (thus confirming that chloroplasts swimming in a protoplast suspension possibly don't evolve oxygen)
...ice: same as fresh, activity rapidly decreasing after 20 min
...ice and then 4 h in RT: suprisingly the activity is similar to the fresh protoplasts. No decrease in oxygen production
The protoplasts are suspended in a 0,5M sorbitol buffer with the common additional salts (MgSO4, KNO3, KH2PO4,...) and I'm using barley.
It would be nice if you can replicate my experiment to confirm it in wheat.
Hope this was helpful.
Regards,
Luca
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