I would suggest embryo culture in MS medium fortified with kinetin, IAA, IBA, TDZ. I think you need to grow them using auxin and kinetin/zeatin in different promotions. Do you subscribe Journal of Plant Tissue Culture and Biotechnology. My email address is: [email protected]
http://www.baptcb.org
Dear Nasrin,
Like true seeds, somatic embryos also need good food reserves to grow. So, as suggested by Younes, transfer the cultures after SE inititiation onto proline or even arginine, cysteine enriched media. In the induction medium try replacing half of your sucrose with glucose. When they mature you need to remove auxin and grow on basal media. Adding GA3 will help at this stage.
One recent publication on the theoretical aspects and another on maize somatic embryogenesis that address your problem are attached. I could have attached some more helpful papers if you mentioned the species that you are having problems
Good luck!
Ranjith
I think you must break its sleep that related to seed dormancy of natural plants!
Sometimes if u take too long to change the media of the culture (subculturing) that also lead to non germination of embryos. In many research articles, the germination medium for somatic embryos is different from its induction medium. Further use of inositol phosphate and as suggested by yonous add Caesin hydrolysate (CH) nitrogen source in 500mg/l conc along with your usual medium.
Some times cold treatment promotes germination of embryo as shown by Late Professor L.F. Randolph in Iris species and hybrids. Hope it will work.
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