In principle necrotic tissue is dead. >But often in tissue culture you see oxidized tissue which upon addition of Ascorbic acid or other antioxidants could be recovered.

In principle necrotic tissue is dead. >But often in tissue culture you see oxidized tissue which upon addition of Ascorbic acid or other antioxidants could be recovered.

My greeting,

plant tissue  is necrosis in tissue culture, because of Increasing the concentration of ethylene gas in containers of culture, as a result of the closure of lid strongly.

As previous replies,  necrosis is dead tissue and can't be rescued.  If however plant tissue is simply brown around the edges and the actual stem still shows signs of being green  you can work with this by using ascorbic acid in the media to help alleviate the problem.

 http://www.arpapress.com/volumes/vol15issue1/ijrras_15_1_02.pdf

Plant necrosis is not happening most of the plants if using correct media formula some plants habit is leaf necrosis like woody plants but it is not giving any harm to process only this need to be removed that part and do subculture or multiplying.If sudden happen  use first half MAJOR + others as full MS salt concentration may happen to this case so concentration with your media PH too.

Hi Maree,

According to the paper you suggested, ascorbic acid tends to decay in culture media and gets oxidized by reactions catalysed by Cu (II) and Fe (III), both of which are components of Murashige and Skoog media; and activated charcoal, being a potent adsorbent would adsorb media nutrients along with the toxic substances produced in the media. 

So, how adding ascorbic acid or activated charcoal would demote necrosis in plant tissues?

Dear Swati

Necrotic tissues are dead  tissues and it is irreversible process. If in your cultures, there are some live tissues, subculture it into new culture medium for further growth. U can use ascorbic acid up to 250-100 mg/L or activated charcoal up to 500mg/L.

Accumulation of Higher concentration of ethylene gas inside test tube and/ or culture vessels, are also responsible for increasing necrosis. Try to avoid to more tight or closure of vessels lid and or also avoid wrapping culture bottles caps with plastic films or u cal use ventilated caps also. So ethylene gas can easily release from bottles.

Better way, keep on monitor your cultures for phenol leaching. Once u feel necrosis is started, please do subculture soon.

Regards

Rajesh Pati

@ Swati,

1. What Maree mentioned is referred to the 'oxidation of phenolic compounds', which is due to the injury of the tissues. Such as edge-cut explants on the medium, which causes 'browning' on the surrounding of the cut edges and the medium the tissue was laid on. For browning, the paper attached by Maree provides several methods to reduce its impact. This is different compared to the concept of 'necrosis'.

2. Are your plants 'chlorosis' or 'necrosis'? I attached a paper for you about chlorosis vs. necrosis (see attachment).

Definition of chlorosis: abnormal reduction or loss of the normal green coloration of leaves of plants

Use cotton plugs to facilitate aeration to avoid ethylene accumulation, more over try to culture in auxin fortified medium.

Regards

Thanks Dr. Yuan, Nissar and very special thanks to Dr. Rajesh Pati for all the help and suggestions in understanding the problem.