Antibiotics for tissue culture

John Korte kortej at css.orst.edu
Mon Dec 9 12:10:48 EST 1996


I recently received this message in response to my question (and was asked
to post it since the involved company was having difficulty doing this)
about antibiotics for tissue culture:
I haven't tried this product and have no association with the company etc.,
etc...
John Korte
Dept of Crop Science 
Oregon State Univ


PLANT CELL TECHNOLOGY'S PRESERVATIVE MIXTURE (PPM)


	Plant Cell Technology mixture (PPM) should be used in plant tissue culture
labs
as a substitute for commonly used antibiotics such as Carbenicillin and
Cefotaxime.  PPM
prevents the
germination of both bacteria and fungi spores, it is heat stable and
therefore can be
autoclaved with the media.  PPM is less expensive than the above
antibiotics and makes
it affordable for much wider use.  The recommended dose of PPM is non-toxic
and does not
impair in vitro seed's germination, callus proliferation and callus
regeneration.  We
are presently testing the effect of PPM on protoplast culture and against
Agrobacterium
tumefaciens.

	For routine tissue culture work the addition of PPM will substantially
simplify
the working procedures as follows:

	1)	Media can be dispensed outside the laminar flow hood (LFH) exposed to
the ambient air.  The plates should be covered soon after agar
solidification.  In the
event that media dispensing is done by a pump, we recommend passing
autoclaved hot water
through the hoses prior to and after media dispensing.

2)	Heat sensitive or heat stable liquid media, does not need to be
sterilized by
Millipore filters or autoclaved providing that it will be stored in sterile
containers
and that the stock solutions are not contaminated.

3)	Working in the LFH the utensils (forceps or scalpels) do not need to be
flamed.
 They should be periodically dipped in 70% alcohol.  The LFH's do not need
to be
certified and the work can be done as well outside the LFH's on a clean
surface for a
period not exceeding one hour.

	PPM has certain limitations.  It is ineffective when exposed to high
density of
bacteria or fungi spores found on plant seed coats.  For in vitro
germination, the seeds
should be conventionally surface sterilized with bleach.  However, after
sterilization,
the seeds can be rinsed under tap water in a non-sterile strainer and left
to dry in the
LFH.  Protoplast isolation solution should be sterilized mechanically
through Millipore
filters with the PPM.  If the utensil ends have touched active bacteria,
fungi culture
or otherwise suspected of being contaminated, they should be sterilized by
autoclavation
or by use of an electric heating element.

	In conclusion, PPM most definitely will facilitate the work in any plant
tissue
laboratory.  However, conditions in each lab may vary.  It is advisable
that the staff
should follow the above guidelines and thus find out for themselves how
much "freedom"
they achieve by using PPM.

	PPM is manufactured in the labs of Plant Cell Technology, Inc.  PPM is
patent
pending.  The recommended dose is 0.4 - 1.0 ml/liter.

PPM was developed by a new biotechnology company called PLANT CELL
TECHNOLOGY, INC.
The company is currently providing samples to labs at no cost for testing.

PLANT CELL TECHNOLOGY, INC. has not yet begun to introduce the product to
the market.
However, the company believes its pricing will be about 1/5th of the price
of 
antibiotics.  For example, if a plant tissue culture scientist currently
spends $10 on
antibiotics per liter of media, PPM will probably be sold for $2 per liter
of media.
This would allow for a much greater use and application for the product.

For more information, please contact Martin Kalin at (202) 463-0904 ext.
134.  The
address is PLANT CELL TECHNOLOGY, INC. 1920 N Street, NW Ste. 750
Washington, DC 20036.




More information about the Methods mailing list