I'm trying to isolate tomato protoplasts for gene editing, and most references that I've found suggested to keep the plants grown the culture room to be kept in the dark for 3-4 days before being cut and exposed to enzyme treatment. Any idea why?
Hi,
I think that earlie (when the technique was optimized for preparing protoplasts) everybody worked with a reporter gene gus. To diagnose its expression, it was necessary to get out of chlorophyll as soon as possible. Indeed, the blue color (as a feature of this reporter geneexpression) is hard to see when the cells is colored green.
Hello
Probably one reason was also an improvement of protoplast production by weakening the cell cements and a decrease in chloroplast volume due to etiolating. Another reason could be that étiolating make the future cell dedifferentiation easier by lessening the photosynthetic function.
Hello
I think, One of the possible reasons was a decrease in chloroplast volume due to etiolating so that étiolating make the future cell dedifferentiation easier by lessening the photosynthetic function.
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