Re: Tissue culture questions

Andreas Wistuba (A.Wistuba@DKFZ-Heidelberg.DE)
Sun, 9 Jan 1994 13:26:09 GMT+1

> Date: Fri, 7 Jan 1994 17:07:45 -0800
> Reply-to: <>
> From: Rick Walker <>
> To: Multiple recipients of list <>
> Subject: Tissue culture questions

Rick, you asked for informations regarding tissue culture media. I
hope the infos I added to your mail is of value for you or somebody

> Now that I've gotten some success, I'd like to try some other plants.
> After gleaning through the archives, this is what I came up with for
> recommended media (these are mostly due to Jan S.):
> _Pinguicula_
> Carroll's media (ICPN v11 n4 12/82 pp. 93-96)
> Murashige-Skoog media diluted 1:5, sucrose 20g/L
> Terrestrial _Utricularia_
> Carrol's media
> Aquatic _Utricularia_
> Carroll's media (no agar)
> Pringsheim & Pringsheim media

> _Sarracenia_

I use 0,66 x Knudsen C* with various concentrations of BAP and IBA
for multiplying and rooting. I also used 0,16 x MS but I like Knudsen

> _Nepenthes_
> Knudson C media

I use the same media as for Sarracenias (mostly Knudsen C*) and I
add 0,2 mg/l - 2 mg/l BAP for multiplying. I feel that a special rooting
medium is not necessary as the plantlets root easyly when
transplanted to soil. I also used Andersons medium (Sigma) and it
worked very well also. N. ephippiata seems to like it.

> _Drosera_

They grow well on 0,66 x Knudsen C*. I'm still experimenting to find
an optimal multiplication medium.

> _Cephalotus_

Sigmas modified MS (1/2 x macro-, 1 x micronutrients) works great for
me (M 0153) with various concentrations of BAP and IBA.

> _Dionaea_

see publications.

> _Byblis_

Just use the same medium as for Cephalotus. It works for all B.
liniflora types and mostly for B. gigantea. However sometimes I have
problems with vitrification in case of B. gigantea.

> For solid media, use 8-12 g/L agar.

*= You may add 37,26mg/l Na2EDTA and 27,8 mg/l FeSO4 x 7H2O.
I always add the MS-vitamins to the Knudsen C medium.

> Finally, is their any truth to the persistant rumour that _Nepenthes_
> been successfully cultured from tendrils? If so, how is it done?
> etc...)

There is a Diploma thesis done on this by Mr Thilo Schmidt-Rogge some
years ago. He managed to callus pieces of tendrils but he did not get
plants from tis callus. Several people including myself have tried to
reproduce his experiments but we did not manage even to get callus.
Even more the explants died rapidly within a few days while controls
on various other media stayed alive for months. I then tried to find a
medium which alllows callus-production from tendrils and ended with
media containing Thidiazurone in various concentration as well as a
whole set of other hormones but all I managed to produce was callus from
tenddrils and leaves, but I failed to getting plants from this callus.
I do not know who wrote this ad in CPN some years ago but I wonder
whether somebody managed the problems. I'm in doubt a little regarding
the diploma thesis mentioned as one of its major conclutions was that
Nepenthes are absolutely dependent on amino-acids in the media. I've
produced thousands of Nepenthes in vitro non of which ever was fed with
amino acids. Maybe best I do not tell them about their needs.

Some literature which might be of interest:

Adams et al. 1979. HortScience 14(6):701-702.

Hutchinson 1984. Scienta Horticulturae 22:189-194.
Beebe 1980. Bot. Gaz.141(4):396-400.
Parliman et al. 1982. J.Amer.Soc.Hort.Sci. 107(2):305-310.
Parliman et al. 1982. J.Amer.Soc.Hort.Sci. 107(2):310-316.

Janssens 1986. Med.Fac.Landbouww.Rijksuniv.Gent. 51(1):61-66.

Bunn 1985. Australian Horticulture. 83(5):103

Rathore et al. 1991. J.PlantPhysiol. 139:246-248.
(They multiply N. khasiana. You better forget about the results of this
paper in use for other Nep-species as N. khasiana seems to be the only
species hardy enough to survive the extremely auxine rich media they
describe. I tried them on other species and my plants endded as a bunch
of roots coming even from the leaves!!!!! However I think this could be
a hint that tissue from Nepenthes leaves is indeed able to regenerate
non-leaf tissues. (I'm thinking about this tendril project I'm still
working on))

There are a few more papers about Drosera and Cephalotus I did not
mention above.

All the best

Andreas Wistuba