Date: Wed, 16 Aug 2000 22:29:56 +0200 From: "Guy Van der Kinderen" <Guy.VanderKinderen@rug.ac.be> To: cp@opus.hpl.hp.com Message-Id: <aabcdefg2680$foo@default> Subject: Re: TC of Pinguicula?
Dear Miguel,
> I found some info on TC of temperate pings, but I'd like to know what
> sterilisation protocols people use successfully for the seeds of these,
and
> leaves (summer or winter) of Mexican Pings.
I've cultivated once Ping. lutea in vitro from seeds and these were
disinfected with 5 prct. of basic calciumhypochlorite with the addition of 1
prct. of Tween 80 (or 20, or another surfactant) for 15 to 20 minutes. They
were subsequently rinsed five times with sterile water. In temperate
Pinguicula, you could also use gemmae to start tissue culture from the
inside meristem. In this case, the gemmae can be surface sterilised with
hypochlorite or alcohol, and the meristem is subsequently removed under
sterile conditions.
> I understand that carnivorous ping leaves are hard to sterilise, as they
> brown very easily. How about non-carnivorous leaves? Being thick and
> fleshy, they should be easier to sanitise...
I have no experience with leaves to start in vitro cultures, but the
disinfecting technique is much more laborious. To produce a cloning line, I
think it would be easier to start raising plants in vitro from seeds, and
then use the germ-free plants for further cultivation from leaf-tissue.
Kind regards,
Guy
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