Maybe crazy tissue culture experiment with Nepenthes seeds

[BigBella]

4 seeds of nepenthes maxima "giant wavy" germinating on sphagnum. Nothing in TC yet. No fungus, no germination. But some seeds seem.... swollen.

Several other nepenthes seeds are also starting to germinate on sphagnum - very early hints of green.

Nothing in TC

Be patient with the seed in TC. Sometimes, it is helpful to keep the cultures in the dark for a period of 7-10 days after sowing. That has proved successful for me in the past; and has generally "sped" germination by about ten days, over those under light.

Also, wasn't 50% MS used as germination media? That can be an issue; it's a bit strong. In the future, try 1:4 or 1:3 and soak the seed in distilled water overnight before sowing . . .

 

[Vidyut]

Yes. Being patient as long as it takes to see germination unless there is contamination - in which case I am prepared to replate the seeds if I can. The delayed germination may also be because my media is not how they want it given that I've used too much MS.

 

[Vidyut]

Most of the seeds on sphagnum had at least one seed germinating slightly over 2 weeks. It is still one day short of 3 weeks.

No action in vitro, but not worried yet. From all accounts it seems to be the sphagnum seeds that are on the early side.

 

[Vidyut]

I had a minor accident that led to one of the TC containers falling to the floor and cracking open. I took the seeds out and rinsed them and have them soaking in water to sterilize and put in TC again.

Gave me the opportunity to look at the TC seeds and medium with a magnifying glass. Still no signs of contamination. I checked the pH, but my pH meter appears to be dead. According to strips, the pH is between 5 and 6 - which seems fine.

Other containers as usual I think - no contamination, no germination, though I haven't looked at them too closely yet, while I was busy rescuing these seeds. This is now getting boring.

 

[Vidyut]

I found some more seeds I'd set aside. I'll do another batch of TC tonight and this time I'll keep them in the dark for a week or so like @Big Bella suggested. Other than these seeds - I guess it is too late to keep these in the dark now.

 

[Vidyut]

Update: fail

I forgot about the seeds for a long time and somehow the media in the containers dried out! It still looked transparent, so I couldn't make out from looking, but when I picked them, the media seemed more solid than I expected and I looked more closely...

I think the containers may be slightly permeable or allowing humidity to escape or something. There was no sign of contamination (except for one container which I had dropped) all through, so nothing seems to have got in. I suppose if I had spotted the problem early enough, I could have added water or put them in fresh media or something, but by the time I noticed the problem, the media had already caked and shrunk and separated slightly from the walls of the container...

Edit: It wasn't dry, but it was quite thick and I assume must have dehydrated the seeds - which hadn't germinated. After I threw away everything, I regretted it thinking that I could have at least experimented further with one or two of them...

But the lack of contamination is promising. I am going to try this again with better containers.

 

[bluemax]

'Strikes me as the right attitude. As long as you learn useful things you succeed. And as long as you continue to try you will succeed fully.

 

[Copper2]

Try it with something else first. There’s no point using seeds that usually take 3 months to germinate because it’s 3 months wasted if your media kills the seeds. Use something like lettuce seeds, get the media dialed in, then sprout the nepenthes, and you’ll make TC history

 

[Vidyut]

Try it with something else first. There’s no point using seeds that usually take 3 months to germinate because it’s 3 months wasted if your media kills the seeds. Use something like lettuce seeds, get the media dialed in, then sprout the nepenthes, and you’ll make TC history

This is actually a good idea! I'd been busy for a while with son's health, etc. Finally getting a bit of time. I'll try doing this soon, and if it works well, will try with nepenthes seeds again.