Re: Brian Quinn's Stage I Contamination of Nepenthes Leaf Explants

Peter Cole (carnivor@bunyip.demon.co.uk)
Sun, 02 Jun 1996 12:15:11 GMT

...
> I am curious if you there is a published or unpublished protocol that you
> are using for regenerating adventitious plants from Nepenthes leaf
> expalnts? I would be happy to know of this, if you care to share. Any and
> all comments, objections, and /or observations are welcome to my basic
> generalities of Stage I disinfestation.
Lines: 30

I always understood that Nepenthes do routinely suffer from systemic
(and symbiotic?) fungal infestation pretty much throughout the plant.
Logically then the best way to successfully TC these would be from
IV cell lines derived from seed, or perhaps less practically from
micro-meristem explants in the same manner as deriving virus-free
cell lines in other plants (though I've only ever seen references to
this concerning agricultural crops and orchids.) Has anyone ever
tried pollen cultures with Nepenthes? I too would be very interested
if anyone knows of a protocol that works with Nepenthes leaf tissue.

In a similar vein, does anyone have a good protocol for Heliamphora
sterilisation? My attempts have met with 100% fungus, with 10%
household bleach (hypochlorite,) soaking for times up to 25 minutes.
The effect on the leaf tissue seems markedly detrimental even before
the fungus swamps the inoculum! Sterile rinsing and transfer chamber
seem evident from my good success with several other genera, but
Heliamphora so far eludes me. FWIW I have found 10 mins to be
enough for D.capensis, filiformis, etc. and Genlisea hispidula, and
15 mins for Cephalotus, but I don't know the original strength of the
bleach because it's not marked on the bottle.

Happy cloning,

Peter

snail:Peter Cole,17 Wimmerfield Cr. :mailto:carnivor@bunyip.demon.co.uk
Killay, SWANSEA SA2 7BU, WALES :http://www.angel.co.uk/flytrap/index.htm
vox:+44 1792 205214 :ftp://www.angel.co.uk/pub/flytrap