I had carried out a micropropagation work of Dendrobium nobile using in vitro raised explants nicely before and from the excised explants shoots proliferated nicely,. But when I tried the same with in vivo explants from wild I am getting hardly any shoots getting proliferated however PLBs are coming? what may be the reason? I used mercuric chloride, detergents, Sodium chloro hypochloride and alcohol for sterilizing explants? Can they affect the regeneration process? I am enclosing our previous paper which will help you explaining my query.
There are several factors.In vitro plants are highly adopted for culture condition.Endogenous hormone level and tissue orientation is different in vitro plants. Secondary metabolite content is more in field plants.Further strelization procedures also inhibit the regeneratin ability of explants which are collected from field plants.
Please go through my recent articles published in Plant Cell Tissue and Organ culture Vol 112 (3): Page:387-393 (2013), In vitro cellular and Developmental Biology Vol 50(6) Page:752-759 (2014) & The Scientific World Journal . DOI:org/10.1155/2014/652919
I hope these articles may be help you in some extent. For more details you can refer the book titled "Plant Tissue Culture" by Bhojwani and Dantu (Springer 2013).
Thiadiazuron (TDZ) is the best growth regulator, it increases the accumulation and synthesis of purine cytokinins as well as promoting the conversion of adenine to adenosine. It highly influences the metabolism of endogenous auxin and alter the auxin-cytokinins ratio within the plant tissue. Therefore, your in vitro raised explants were nicely responded for this growth regulator and produced multiple shoots.
Laloue and Pethe (1982) Plant Growth Substances, Academic press, London.
Bespalhok and Hattori (1998) Plant Cell Reports, Vol 17:pages 870-875.
Saxena et al., (1992). Planta, Vol 187: Pages 421-424.
Dear Shajahan, yes. these are the papers. three cheers for research gate.
Dear Bhattacharya¡Thank you¡I would like congratulate you for wonderful findings in Dendrobium nobile[Elsevier>METAGENE].I wish you to keep up the good findings in future also.BEST WISHES¡Shajahan
TDZ is basically defoliant when used on plants growing in natural environment. The in vitro grown explants are grown in a protected environment to the extent that they get glucose/ sucrose as readily available carbon source. The metabolism of the tissue grown in vitro is entirely different (that is why the plant-lets need to be acclimatized when they are transferred to soil). Therefore the chemicals which are used in vitro act differently than in vivo conditions. I would like to mention my observation when I use TDZ is that it gives good response when 3 to 4 % sucrose is added.
I had a similar experience with field-derived explants of the East Africa tree Melia volkensii using TDZ. 0.1% Mercuric chloride, 10 or 15 minutes often resulted in 100% no-response and necrosis. However, 10% commercial bleach for 15 minutes resulted in efficient Indirect regeneration from mature zygotic embryos and direct regeneration from mature cotyledons. A good bet would be these two explants.
I think the reason can be due to the product itself because in most of case TDZ doesn't give the good results, like other researchers i try TDZ 0.1mgl-1 in combination with NAA 0.5mgl-1 to induce callus in moringa oleifera it didn't give any callus.
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