to be sure: you are talking about bony/bone callus regeneration or plant callus regeneration? Thanks for elucidation....

Wolfgang H. Muss ji-

plant tissue culture

Hi Sachet Hegde,

How to use a 'histological method' to check whether a plant callus is regenerable or not? Any report?

By the way, in my opinion, there is no effective/standard way to determine whether the plant calli is still 'regenerable'. We can only choose a way to avoid not-regenerable calli in our experiments. For example, we should use NEWLY induced calli for plantlet regeneration. We should avoid using a long-term tissue-cultured calli for plantlet regeneration, because many studies have shown that 'regeneration ability' can loss for those long-term cultured plant cells.

Yuan-Yeu Yau ji, thank you for your valuable suggestion... I know that long term calli must be avoided.. coz I am not getting any type of regeneration from any type of callus, thats why I am asking these things.

Hi Sachet Hegde and Yuan-Yeu Yau,

Usually, the regeneration ability of a plant tissue culture varies widely among the different species and even between different genotypes within a same species. Also, the effect of time in culture depends on the genotype and on the interaction of the genotypes with the media composition. So, I personnaly consider that the best way to verify whether the plant calli are still "regenerable" is to furnish all requirements for plant regeneration: light, temperature, nutrients and adequate hormone composition in the culture medium. Sometimes, even with all environmental factors furnished, only roots or shoots are formed... but not complete plants.

Hi Antonio Fluminhan,

"Sometimes, even with all environmental factors furnished, only roots or shoots are formed... but not complete plants." For this, I still consider they are 'regenerable' calli. It might be just because that the protocol is not optimized and the ratio of hormone added are not optimum enough to warrant the rgenerants developing into wholesome plants.

One situation is that the calli would quietly sit in the plates without anything happen. They won't die, but just sit there for as long as the medium is timely subcultured. In this case, they are most likely 'non-regenerable'. For this, it is hard to predict whether they are regenerable or non-regenerable beforehand---and only time can tell.

@  Sachet Hegde,

Is the tissue culture protocols for this plant species you are working on publisher? If so, you should first follow their protocols, then modify/optimize it to fit your specific varieties by yourself later.

1. Callus should not be in brown colour. Means no secondary metabolites should be there in the callus. 

2. Biotic or abiotic stress will induce 2*ry metabolites. 

3. Callus should be in light cream colour. 

4. Regenerative callus will be friable. 

5. Thin Cell Layers (TCLs) technique is a promising technique for different types of regeneration/ morphogenesis 

* plz go through my publications.

Rajesh Kondamudi  ji,

Thank you for the valuable suggestions.

Surely I will go through the publications made by you.

Dear Fernando Santacruz-Ruvalcaba  20.85 University of Guadalajara:

I feel honored for the recommendation you gave my answer #01.

For others to know: when I replied that moment given topic(s) only was Histology and there was no extension of title and in text of the question: " in plant tissue culture". It is good to see that there were plenty of knowledgable answers. With my compliments and best wishes for success,  WM

Wolfgang H. Muss ji,

that was edited by me afterwareds..sorry for that..

and thank you for your valuable suggestions... if you want to add some more answer some more suggestions.

Dear @Sachet Hedge, thank you for your comment #012 to my Re #011 which is welcomed. The possibility to edit  questions as well as comments/posts and so to correct, add or supplement (lacking / valuable / supporting / new) information is one of the striking & beneficial properties in the ResearchGate Platform. As a former student in Zoology and Botany for PhD (in the 1970ies) I knew about the two possibilities (bone and plant callus) - so I asked frankly but honestly about which 'direction'/condition you were asking....  Since I 'unfortunately' regarding my former profession I followed the human biology (= more 'zoological') / human-medical thread rather than that one directed towards plants and/or plant physiology (the latter I took my final exams in Botany with A-grade)...  I can't comment any further on your plant callus matter / problem...since 1979/1980 I haven't done any (research)work on plants (though reading and recommending specific (esp. methodologic) articles in plant research sometimes on RG.

Finally, I found it wonderful how your question topics were expanded... therefore many specialist will find the / your question, I guess.....Best wishes and regards, -W.M.-

Wolfgang H. Muss ji,

thank you...ofcourse I too from chemistry, botany and zoology (B.Sc) background, but while framing questions I didnot remembered animal tissue culture so I started to write such stupidly without specifying the type. I thank you very much in this regards as because you just reminded me to rewrite/edit the question that I had.

thanks for the wishes too.

Yes, I agree with Mr. Rajesh. One of the indicator of a regenerable callus would be that the callus looks friable (fish-egg type of callus), in which it is usually termed as embryogenic callus. You can search for friable callus image to get a rough idea. Good luck

Dear @Sachet Hedge, you're welcome! 

You need not vilifying yourself in saying "stupid writing [of question title]", since

| Nobody is perfect....! | (naturally including myself!).

It would be interesting if you in the end could summarize how your methodological problem was solved successfully. Best wishes again, W.M.

Dear Sachet,

I did not get what you mean with "regenerative". You mean an increase in size? Or increase in weight? Or you regeneration to plants through direct or indirect methods (organogenesis, embryogenesis,...). There is callus index method for size growth. Fresh and dry weight method used usually for secondary metabolites production.

Good luck!!!

Dear Hussien M. Daffalla ji,

I refered about regeneration of plants through callus...