Dear Rizwan

The induction of embryonic calli can vary depending on the plant species and hormonal treatments. Please provide more information about your experiments so that I can help you.

Seied Mehdi Miri

Respected  Sir, Seyed Mehdi Miri

 I am a student at Fujian Agriculture and Forestry University Fuzhou China with a major in Horticulture.

I am working on Okra vegetable for regeneration through callus, but while callus induction facing the whitish compact non-embryogenic callus, with very low shoot induction. So my question is that :

1: Factors affects the whitish, compact non-embryogenic callus induction?

2: Is there any role of Nitrogen in embryogenic callus induction?

3: Please explain these issues with some previous article references.

Waiting for your kind response

Somatic embryogenesis depends on so many factors such as plant species, explant type, explant age, media composition, growth regulators etc. Unless the detail experiment is known, it is difficult to suggest any modification. Please give the details so that I can help.

Dear Hafiz

The ability to produce embryogenic callus depends on factors such as plant species, type of explants, composition of culture medium, type and concentration of growth regulators. Development of embryos can be controlled by the appropriate manipulation of growth conditions (growth regulators, use of reduced nitrogen from different sources, osmolarity, etc.). The level of reduced nitrogen supplied to cultures is one of the most important factor, which control in vitro somatic embryogenesis. Suitable somatic embryogenesis treatment of any species depends on the culture medium and the concentration and source of nitrogen in the medium. A combination of reduced (NH4) and unreduced (NO3) forms of nitrogen, or amino acids (alanine, glutamine, arginine, lysine, serine, asparagine and ornithine) stimulated the numbers of somatic embryos and proceeded globular embryos to maturity.

Seied Mehdi Miri

Thank you very much ,

Your articles and suggestions very helpful to me,

Thanks again,

Manoj Mishra,

Seied Mehdi Miri ,

Sir i have attached the pictures of my experiment kindly have a look, white compact calluses.

Sir, whats the role of nitrogen in callus or shoot induction? have you experience or references about it then kindly share.

From the pictures, your agar medium looks kind of dry, and cracking. How frequently do you subculture your tissues? How much agar did you use for a liter of medium? I have seen this on my carrot callus long time ago when the agar medium stayed for a long while.

Yuan-Yeu Yau

i use 8% agar ,

What are the plants and treatments that you used to shoot regeneration?

@ Hafiz Muhammad Rizwan,

What did you mean by 8% agar? 8 g agar in 100 mL medium? I think it should be 0.8% (8g in 1L medium).

And how often you subculture your calli? The medium in the first jar (in your picture) is almost dry up?

What is the media copmposition. Is it contain high 2-4D or NAA as the auxin source. Further in the second jar near to your white calli there appears to have some brown calli. Those calli are important and may turn embryogenenic in future so look for those type of calli. Also see that media should not dry. But simultaneously do not subculture very frequently. Some plants need a bit stress before turning to embryogenic.

Respected all,

I too have the same problem with the formation of somatic embryogenesis/indirect morphogenesis but I have tried nitrogen free media and half strength ms and wpm with 0.8% sucrose and some modifications like addition of maltose and agno3 but got no response from the whitish compact calli. so kindly suggest some articles.