RE: 4°C suspended plant cultures

Deanne Bell dbell at fresno.ars.usda.gov
Wed Mar 29 12:07:35 EST 2006


>-----Original Message-----
>I'd like to know about the possibility of preservation of in 
>vitro plant cultures (specially woody plants) at 4°C.
>Nasser Mahna

Hi Nasser,

I don't have any experience with your exact situation, but I have been doing quite a bit with tissue culture lately.  I think that you really have to go with mother nature in regards to what the normal life cycle of the plants are. 

What I have found to work for my deciduous commodity:
*I take cuttings of small stems in the fall (before the rains helps to keep fungal contamination down)
*disinfect and place into liquid growth medium
*Store in cool room, in dark, until the proper number of chilling hours for my commodity has been met.  I imagine you can keep them in some sort of suspended cold storage at this stage fairly easily.
*Move samples to growth room with 16 hours light and correct growing temp
*Buds will break in 2 - 4 weeks.  I can't imagine taking these young tender tissues and trying to keep them alive while at 4°C.

I have had some limited success with culturing shoot tips in the spring when the young shoots are actively growing.

>From your question I think that you have green-stemmed plantlets (from a woody plant commodity) growing on some sort of plant agar medium that you want to cool down or suspend growth at 4°C:

I do not know for sure but my gut instincts say that these are the things to consider:

1) is it a deciduous variety?  they probably would not take well to 4°C cool temps, might force it into dormancy.
2) does it normally live in places that get down to 4°C? then maybe it would be OK for a short period of time.
3) do you have a way to keep it under normal lighting conditions while it is at 4°C? 

When all literary research and intelligent predictions are exhausted, there is always the scientific method of experimentation:  try a few plants at 4°C for 2-3 weeks and then bring them back to normal temperature and observe for a few weeks.

hope this helps,
Dee Bell



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