The growth of my somatic embryos have been arrested in heart-shaped or torpedo-shaped.
Hi Atefe
transferring the embryos in a medium (MS , B5, WPM ... nutritif medium) which contains phyto hormones that promote organogenic and especially the formation of roots
Kinetin stimulates organogenesis
ANA , AIA , 2,4.D for root formation
concentration depends on the species
Best regarde
Try the following by transferring torpedo-shaped embryos by long forceps one-by one onto:
1. Same Media with 1/2 strength (if B5, then use 1/2 B5 or 1/2 MS depending on your media). +
2. Add, IAA + Kinetin + GA (first two 0.5-1 mg/ L) and the last one 0.015-0.05 mg/L etc.) +
3. Reduce sucrose (Carbon source) by 50%. +
4. Increase Agar/ Agarose by 1.5 % to make media more hard/ solid.
Reasons:
1. For promoting lack of nutrient condition and hence photautotrophic growth in light.
2. For growth, rooting, shooting and chlorophyll biosynthesis.
3.To promote autotrophic growth.
4. To allow rooting nicely.
Try conditions 1, 2, 3, 4 independently and also "in combination of all 4". If growth inhibition of embryos is due to "yellowing/ browning of agar media, due to phenolics that are growth-inhibitory and can kill them, then add "charcoal" in the above suggested media to hep them grow!
Best wishes.
depend on the inside hormones of your plant
and try the methods for Dr; Biswapriya Biswavas Misra
It's really dependent from the species! Somatic embryogenesis in monocots is much more difficult and their requiremenst for mediums and hormons are not the same like for the dicots. Somatic embryogenesis can be also genotype dependent in some extent. Do you work with genotype with high embryogenic potential?
Hi,
The optimal medium for spring wheat embryo culture is MS+2-4 mg/l 2.4-D, sucrose 20%, agar 0.7-0.8%
I am studying on genotype with low embryogenic potential and I would like to know general principle of it.
Better you should stop the usage of auxins after torpedo shape embryo formation.In many cases auxins are only useful for to initiate SE induction.Continuous usage of auxin will cause either callus or root formation from induced SEs.Balanced exgenous and endogenous hormone level is essential for further development.So after shape formation you can start from hormone free medium for further development.You need more please refer my recent Curcuma amada somatic embryogenesis papers.
Thank you Dr. Shajahan
I used hormone free medium but was not observed any growth.
Somaticembryogenesis is comletly plant dependent.Your plant is monocot or dicot?Any way,endogenous and exogenous hormone levels,nutrient media and physical factors are also play important role in proembryo development..In some cases, ABA enhances epidermis development. During proembryo stage,they can synthesis their own auxin and polar transport of these auxin also influence its development.So several trial and error experiments are required to achieve the goal.
I hope you working in mocot and my following articles may be helpful in little extent.If you are working in dicot, please refer my cowpea somatic embryogenesis papers.
Article An efficient regeneration system via somatic embryogenesis i...
Article Direct somatic embryogenesis and plant regeneration from lea...
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