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I have recently started to try and produce a cell culture using maize embryo's, initially on solid media in order to gain the callus structures that I need. The problem that I am having is that all the embryo's that I extract and place onto plates for growth seem to then just enlarge and pretty much germinate.
I don't know if this is because the embryo's used for this were too far along developmentally or a different problem. I am aware that there are larger more solid embryo's and then also slightly smaller translucent ones.
- Embryo's used between 1-2mm in length
- Plates are kept in the dark at 25.C
- Using MS media, agar, 2mg/l 2,4-D and MS vitamin addition in plate media.
As far as I know the everything up to placing the embryo onto the plate is fine (extraction, sterilization, embryo axis on media, etc) therefore the only thing I can think of is either something to do with the media composition/the way i have made it or the embryo stage I am using.
I have yet to see anything that looks like a callus of any sort in the images that I have seen, just enlarging embryo's.
(I have probably left out quite a lot of details)
Any help would be greatly appreciated
Dave
