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Nepenthes seed in vitro

Hello

What is the best way to sterilize Nepenthes seeds. I tried dipping them in 70% alcohol then soaking them in 10% bleach solution for 10 minuets then rinsing them off in pure water but that seemed to have killed the seed embryo.

What recipe of MS, PPM and BAP would work best for Nepenthes seed?

Any help is greatly appreciated
-Jeremiah-
 
Never done tissue culture before. Good luck man.
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Try eliminating the alcohol. My guess is it is this step that killed the embryos. When I was working with orchid seed in vitro we never sterilized with anything but 10% chlorox soltion.

Good luck.

Phil
 
I don't think Jeremiah is tissue culturing, but raising the seed in flask, as an orchid breeder would do.
I know one orchid flasker who claims that hydrogen peroxide is an effective replacement for the 10 percent clorox wash. How he dilutes the peroxide is unknown, but claims it is gentler on the seed.
As for agar formulations-have you investigated some of the commercial pre-packaged formulas available for orchid growers...say something formulated for Cymbidiums, Cattleyas or Paphs.
I have no idea what the nutrient requirements for Neps would be. I'll be watching this thread closely to find out. Tony?

Trent
 
When people say 10% chlorox wash, it means that 1 part of chlorox has been diluted in 9 of water? or the final concentration is 10%.
In our part of the world, we have this chorox equivalent called whiteking and in the bottle says it has 4.9 mls of pure sodium hypochlorite per 100 mls of water. In other words, the concentrated stuff is only 4.9%. If we dilute that 10 times, we'll get .49%.

How much sodium hypochlorite is there in pure Chlorox??

Thanks

Gus
 
Very interesting indeed Jeremiah! I would like to try this myself. Keep us updated. Mabey some pics???

Gus,
It's 6% here in the states.

Robin
 
Me too, Robin. I would think there would be a higher germination rate, and more surviving seedlings in vitro as compared to sowing the seed on sphagnum and peat.
Definitely something worth looking into.

Trent
 
Trent, interesting hypothesis, but I think it's a case of six of one, half a dozen of the other. Both have their advantages and draw backs, and the end result after a year or two is often not that big.
 
Yea. I guess the runts will always be the runts. We've sown many Nep seeds, and you always end up with a fraction of the siblings that grow faster and stronger than the rest, and there is a high attrition rate at the compot stage. Only the strong survive-even in the greenhouse.

Trent
 
  • #10
I have some wild seed grown plants (N. burbidgeae and N. stenophylla) I've raised them myself up to now, but would like to know if it would be feasible to give them some Superthrive (for root development) or what I give my immature and mature plants, the Osmocote slow release stuff. Currently they are in high quality sphagnum and just getting regular RO water.

Thanks for any input.
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  • #11
Seedlings are much less tolerant of fertilisers than adult plants. Whereas adult plants will react by not producing pitchers for 6 months, seedlings can drop dead on the spot. Fertiliser can be used in extremely weak doses, but in the case of a species like burbidgeae, I wouldn't be taking any chances.
 
  • #12
Nep G, remember that Superthrive isn't a fertilizer but a hormone, there's no actual nutrient value in it. It's kinda like a snickers bar for your plant, might give it a bit of 'umph' but will still not actually feed the tissues.
I would try a foliar mist of urea free orchid fertilizer such as Gro More which is very easy on plants and is an immediate source of useable nitrogen. It's a bit pricier than regular orchid fertilizers but it's far more useable.
I've been foliar misting with 1/2 strength Gro More on my neps for the past few months because I've been too lazy/busy to buy and distribute a couple hundred crickets every couple weeks. Everyone is growng, flowering and pitchering fine. Even the tiny plants are doing fine with the occasional misting.
 
  • #13
Also note that wild-collected seeds will likely need a longer period of sterilization compared to seed collected from a greenhouse.
 
  • #14
That's an interesting point for discussion. Someone recently posted a message (here?) saying they thought that greenhouse seed had a higher rate of fungal infection. They postulated that is was due to the fact that closed environment in greenhouses was an ideal breeding ground for pathogens.

Any comments?
 
  • #15
A couple months ago, I tried sowing some Nepenthes fusca and lowii seeds in-vitro(probably from same batch offered by the icps). The fusca appeared to be wild-collected as the location data was rather specific. The lowii was labeled as lowii-Trusmadi. I sowed both seeds using the same medium and protocols and I noticed that almost none of the lowii flasks were contaminated. On the other hand, quite a few of the fusca flasks were contaminated(around 50%). These seeds were sterilized with 10% bleach solution +tween20 for around 15-20min. I suspect that wild-collected seeds would contain plenty of fungal spore and bacteria. Diluted MS medium is probably not the best choice for these nepenthes as they are growing very slowly...This medium works much better with Drosera and Pinguicula.
Ideally, the seedpod should be collected right before the pod splits.
 
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