Monday, October 7, 2013

Nice Tips from Sean Bulley

Interesting to find out that rooting eucalyptus micro shoot in a medium sterilized with micro wave shows better result compare to medium sterilized by autoclaved, may be the auxin is lost.

Plant Tissue Culture Research PT WKS Sinarmas Forestry

Comments

  • Lisa
    Lisa Masterson
    Director, Marketing and Sales at Plant Cell Technology
    I've not tried it, but from what I've read the 120deg C AND the pressure are critical. Microwave creates heat essentially by friction at the molecular level. I'm not a physicist or physical chemist, but I would suspect there could be damage/degradation to various compounds. Just speculation. I would have to see comparative proof!
  • Sean Bulley
    Sean
    Sean Bulley
    Plant molecular biologist and biotechnologist
    microwaving generally destroys more nutrients than other forms of 'cooking' so your results are puzzling. Maybe microwaving breaks up the agar a bit more and maybe some of that component has some plant growth regulator promotion?
    Anyway, I have always added my growth regulators after autoclaving.
  • FREDDY PANGARIBUAN Indonesia
    FREDDY PANGARIBUAN Indonesia
    Plant Tissue Culture Research PT WKS Sinarmas Forestry
    Yes Sean Bulley, that's what I mean, I put the auxin after autoclaving, and then pipetted the auxin to the solidified medium. And in order to mixed the hormone with the medium, i have to heat up the semi solid medium for 1 minute 30 sec, then it turned liquid when heated, but solidified again after cooling off. I used microwave oven to do that. Compare to the the hormone autoclaved along with the media, the result is much better. My difficult to root clones produce plenty of roots. My next trial will be doing it with sitokinin, please share your experience
  • Sean Bulley
    Sean
    Sean Bulley
    Plant molecular biologist and biotechnologist
    Hi Freddy, I don't let the medium solidify after autoclaving, just wait till it has cooled to below 60 deg C and then add PGRs and mix. I would certainly never add in PGRs before autoclaving as this will degrade them but it sounds like your post-autoclave 1'30" microwave step doesn't destroy the added auxin too much.
  • FREDDY PANGARIBUAN Indonesia
    FREDDY PANGARIBUAN Indonesia
    Plant Tissue Culture Research PT WKS Sinarmas Forestry
    Yes Sean, I got a good result that way. Actually I did't have the filter sterilization to sterilize my auxin. So that's why I have to cook them again to kill the possible contaminant that might presents in my hormone stock. Thanks for the advice, I need to buy the filter then.
  • Sean Bulley
    Sean
    Sean Bulley
    Plant molecular biologist and biotechnologist
    rather than expensive filter sterilising your can just dissolve your auxin in say 20% ethanol and then it should be sterile.
FREDDY PANGARIBUAN Indonesia

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