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Nepenthes in vitro . . .

Here are a few shots of some highland Nepenthes seeds within a few days of their aseptic germination in 1:3 Murashige and Skoog (MS) media. At the left, the radicle and hypocotyl are visible, just emergent; and cotyledons can be seen, just unfolding, at the right -- and escaping the seed coat below.

Nepenthes sp. -- 9 October 2011
NEP-A.jpg


NEPDICOT.jpg


NEPPIES.jpg
 
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great work david--how long did it take for the seed to germinate from the time you introduced the seed to the media?

also i've been wondering...is it possible to induce a callous stage for nepenthes from seed? i know in regular TC methods, one would place the TC flask with plant tissue into a dark area and rotate the flask on some sort of spinner, which would prevent the cells from differentiating and only focus on dividing and absorbing nutrients from the gel... would be great if we can obtain multiple plants out of seed instead of just one.

thanks for sharing. it's always great to see how the pros do it...
 
Very cool
Amp what up wit yo avtar
 
how long did it take for the seed to germinate from the time you introduced the seed to the media?

also i've been wondering...is it possible to induce a callous stage for nepenthes from seed? i know in regular TC methods, one would place the TC flask with plant tissue into a dark area and rotate the flask on some sort of spinner, which would prevent the cells from differentiating and only focus on dividing and absorbing nutrients from the gel... would be great if we can obtain multiple plants out of seed instead of just one.

The bulk of the seed took about twenty-seven days in the various media before germination was seen (courtesy of a magnifying loupe); and, yes, it is possible to induce callus just by tweaking the the various plant growth regulators without the use of either darkness or a spinner. A media containing a large amount of 6-benzylaminopurine (BAP) is usually sufficient to do the job . . .
 
Can't wait for those EB seeds!
 
lol!! BigB...I sometimes wonder....shouldn't you be drowing in neps and helis by now? :p lol! Beautiful pic btw...and thanks for sharing. Not often do we see a nep germinating. :D
 
Here are a few shots of 2011 Nepenthes macrophylla and N. lowii within days of germination. They're certainly nothing to look at for the time being; but it was encouraging, nonetheless, to see that the seed was viable -- and that I knew what I was doing, in an offhand way . . .

Nepenthes macrophylla -- 18 October

MAC-1.jpg


Nepenthes lowii
LOWII.jpg


Here is a representative N. macrophylla from the 2009-10 seed batch:

MAC-B.jpg
 
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ohhhhhhhh me like

:)
 
Awesome, dude. That's really cool.
 
  • #10
Quite impressive work David!

They are germinating very well in your care.
 
  • #12
Very well done! They look really promising. I you don't mind answering, for how long and at what concentration did you disinfected the seeds?
 
  • #13
Very well done! They look really promising. I you don't mind answering, for how long and at what concentration did you disinfected the seeds?

For the last several years, I have only used solutions of PPM (plant preservative mixture) and MS salts to sterilize seed -- particularly Nepenthes, which seem far more susceptible to bleach and / or alcohol damage than most other genera (and is the only consistently-successful method for sterilizing Cephalotus seed that I have found). I alternate between four and eight percent solutions mixed with 1:3 MS salts (just dissolved in distilled water without sucrose, PGRs, or pH adjustment). The seeds, in filter paper envelopes, spend upwards of three to four hours and are plated without rinsing.

There is a blurb on the ICPS site which goes into some detail about the use of PPM and refutes the popular idea that it inhibits germination:

http://icps.proboards.com/index.cgi?action=display&board=tissue&thread=5073&page=1
 
  • #14
Oooh *golf claps* well done, sir!
 
  • #15
Here is a shot from this morning of a culture shown earlier in this thread. They are seed from Nepenthes macrophylla, sown on 9 September; and they currently seem to be enjoying themselves in a "spa treatment" of 1:4 MS with 1 ml/L BAP . . .

Nepenthes macrophylla -- 14 November
MAC2011.jpg
 
  • #16
OMG Big Bella I am green with envy. Fantastic work. I am very interested in the processes you use for germination in sterile media. I have a very good sterile tech, but never took chemistry in college for my nursing degree. So mixing the agar plates and getting the right PH is still new to me. I made one attempt at growing orchid seeds, but figured I would wait till I had a better place to work in and could keep the flasks under lights.

Maybe one day I could pick you brain and get some pointers on the processes used?
 
  • #17
wow great work :-D, but your messing up da smart curve here ! LOL
 
  • #19
Babies are so cute no matter what Kingdom. Congrats.
 
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