i have 2 years old callus obtained from cotton. eventhough im not observing any embryo stages im suspecting somatic embryo formation in callus. but i don't know how to confirm that. any suggestion for this?
when the calluses turned intro brown color or dark brown color then the callus are called somatic embryos. The somatic embryos are usually formed after transferring the calluses to appropriate media and placed in dark culture room.
@ Shiwani,
Are those somatic embryoids in the color picture in your attached document from cotton?
If you really don't see any embryo stage you may proceed with a biochemical and/or a histochemical evaluation of your callus culture, i.e. staining with toluidine blue. Some results for this test could give you answers if you are or not in the right way.
You can tell somatic embryos from the general morphological appearance. You may need a x 5 hand lens or view the callus under a dissecting microscope to check for them. If you see globular, heart-shaped and/or torpedo structures then you got them!
See morphological images in: file:///C:/Users/COMPAQ/Desktop/zhang_2_6_2009_259_268.pdf
But always, the best evidence is histology. You see the same shapes but this is more confirmatory.
For histology of cotton see images in Figure 2 in this paper:file:///C:/Users/COMPAQ/Desktop/psb-8-e26762.pdf
All the best.
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