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Fingerprinting in Tissue Culture
#1
How can one use finger printing in tissue culture to ensure genomic consistency, for a commercial lab that is?
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#2
Which are the basic protocols in micropropagation of Eucalyptus, Camellia sinensis and Coffea Arabica of initiating in vitro considering phenolics production? Conditions for culturing other than initiating in darkness at 25 degrees Celsius.
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#3
Hi Candy,

I assume you want to avoid the production of phenolics (as they lead to necrosis)?

For Eucalyptus, as read in many papers, subculturing seems an answer (though not a perfect answer) to phenolics.

A useful review on  Eucalyptus micropropagation suggested following to avoid phenolics in tissue culture of Eucalyptus (pasted verbatum):

  1. Selection of actively growing young explants.
  2. Use of sterilization procedures that do not severely damage the explant tissues. This may require transfer of donor tree material into a controlled environment, establishment of higher standards of sanitation (for example fungicidal sprays) and partial rejuvenation of the material by grafting or rooting of cutting.
  3. Washing sections of source material in running water for at least 1 h before sterilization.
  4. Soaking explants in sterile water after sterilization, before placing on medium
  5. Soaking explants in antioxidant solutions after sterilization, before placing on medium
  6. Including antioxidants such as polyvinylpyrolidone and ascorbic acid in the medium. Although activated charcoal has been used to adsorb toxic or inhibitory substances formed by callus explants, it was found to inhibit callus formation on various Eucalyptus explants when added to media at 8 g/l
  7. Placing cultures in the dark for an initial period of 7 to 14 days
  8. Frequent subculture during the initial months
  9. Using explants from established shoot cultures

Refer the review here:
http://treephys.oxfordjournals.org/conte...5.full.pdf


I am sure some of these would provide answers to Tea and Coffee micropropagation as well. Hope you went through the very old paper of Koretzkaya?

If you find some other suitable solution for dealing with phenolics, do let me know.
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#4
(06-22-2015, 08:54 PM)CandyWangui Wrote: How can one use finger printing in tissue culture to ensure genomic consistency, for a commercial lab that is?

Can you elaborate on your question a bit? Sorry couldn't get the jest of it..
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