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Discuss all micropropagation related topics here.

Moderator: Matt

By rellenburg
Posts:  241
Joined:  Fri Aug 10, 2012 2:57 am
#162034
I understand ;) Thanks for all the help you do give. I will continue to refine my process and equipment, and hopefully I'll be able to help others get started as you have. You could at least post some pics or something regularly on here to help keep me motivated though. :lol: Fortunately I have had some success.
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Image
By rellenburg
Posts:  241
Joined:  Fri Aug 10, 2012 2:57 am
#162039
Oh yeah, about the greenhouse thing... I never even considered that. I only got my first cp around September of 2012 and plan to build a gh in the spring. I'll give it another go in a few months. Thanks again.
By Frankco
Posts:  14
Joined:  Thu Jan 14, 2010 12:42 am
#162053
I had to go check it out. Eeeek! 45 minutes in 10% bleach for Nep seeds. Nice little white sticks after that.

Anyway, you can see the world-of-carnivores.com by using the Wayback machine. Hardly anything disapears from the web. Well some photos do but the basic webpages are usually there in different past versions.

Main page from Dec. 2008
http://web.archive.org/web/200805091356 ... vores.com/

Nep sterilization method also from Dec. 2008
http://web.archive.org/web/200807050128 ... nthes.html

Frank
By cvlover
Posts:  447
Joined:  Thu Apr 23, 2009 4:59 pm
#162076
Frankco,
We see ton of pictures of your CPs. Which sterilization do you prefer for nep and heli : PPM or NaDCC?
By Frankco
Posts:  14
Joined:  Thu Jan 14, 2010 12:42 am
#162082
Hey cvlover,

I haven't worked much with Heli. Mostley Neps.

I just do what works for me. I'm sure there are other methods that work just as well and maybe better. One thing I do know is that if I say "this is the best way" I'll be proven wrong tomorrow and probably by myself: -)

I was lucky to enough to be able to experiment for a couple of years with seeds that were handled in pretty the same way when harvesting. The seeds were very clean to start with and the plants were grown in a greenhouse. Also the seeds were fresh and I knew the exact harvest date. So, that took out many of the variables that mess things up when trying to figure out what works.

Those seeds were all lowland species and this is what I found out works best for them/me, emphasize “for me”.

The highest germination rate and ratio of clean seeds was from a 12 hour soak using 380mg/liter of NaDCC (99%) with 10 grams of sugar and few drops tween 20. Contamination was very rare. With most batches there was none. Again, those are about the cleanest seeds one could get. The pods were unopened at harvest time and for a while the pods were even treated with isopropyl alcohol before shipping.

With seeds off of eBay the results vary a lot. Generally, those seeds are very “dirty” and you can’t always be positive of the harvest date. Throw in how each species handles NaDCC, how the seeds were handled before they are shipped, was it dry or rainy when harvested, were the pods partially opened when harvested along with questionable harvest dates, it’s no wonder that someone starting out with those seeds has hard time figuring out what works.

There’s a very good chance that 380mg/liter of NaDCC for 12 hours will not do so well with seeds off of eBay and you have up the time or adjust the amount of NaDCC higher or, lower the amount of NaDCC with more time. Plus, adding 10 to 15 minutes in 3% H202 is usually required for seeds off of eBay. With my last batch of seeds off of eBay, a 4 hour soak in distilled water with 10 grams/liter of sugar, then a 12 hour soak in 380mg/liter of NaDCC, plus 15 minutes in 3% H2O2, worked well with good germinate and low contamination.

If I get a lot of contamination from seeds off of eBay I know that the seeds were not handled or harvested in the best way.

With side by side tests the germination rate with 8% PPM + 1/4th MS for 2 hours was considerably lower than NaDCC. Maybe this is not an optimal amount of PPM and the amount of time was not optimal. So, take that with a grain salt. Also I was testing very clean, lowland seeds. The table may turn toward PPM with other types of seeds and less clean seeds. I’m not sure. Bigbella has very good success with it and I’m sure has worked with more types seeds than I have.

Disinfecting Nep seeds.
Image

Frank
Frankco, Frankco, Frankco and 1 others liked this
By cvlover
Posts:  447
Joined:  Thu Apr 23, 2009 4:59 pm
#162086
Your lab looks good Frank. We want to see more pictures. Thank for information.
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By Matt
Location: 
Posts:  21111
Joined:  Mon Apr 21, 2008 11:28 pm
#162093
rellenburg wrote:I will continue to refine my process and equipment, and hopefully I'll be able to help others get started as you have.
You will. Just keep trying new things and taking good notes of the results. Learn what works and what doesn't. Eventually you'll figure out a few small things to add into the process that will increase your success rate.
rellenburg wrote:You could at least post some pics or something regularly on here to help keep me motivated though.
Hah, I should post photos more often. This time of year, there is nothing but jars full of tissue as all of the initializations from the spring are now very grown out.
rellenburg wrote:Fortunately I have had some success.
You're well on your way then. After the first success, just keep repeating what you did to get that success and try slight variations on it to see if the percentage of successes increases. You'll eventually develop your own process that will give you a good success rate. And great job being so successful so quickly. It took me a very long time to get my first strike with a leaf.
Frankco wrote:I had to go check it out. Eeeek! 45 minutes in 10% bleach for Nep seeds. Nice little white sticks after that.
Great to see you on the forums Frank! And yeah, most of the sterilization protocols on world-of-carnivores.com are far too harsh in my experience, resulting in dead seeds or tissue.
rellenburg wrote:Good to see you visiting Frank. You are the other teacher that I owe many thanks to!
I second that! Those videos you published on YouTube a few years back went a long way to getting me started in TC. I can't thank you enough!
By Frankco
Posts:  14
Joined:  Thu Jan 14, 2010 12:42 am
#162097
Thanks Matt. I really appreciate the warm welcome.

I enjoy reading the posts. There's a lot to learn here. This something where one never stops learning new things :)

Thanks again,
Frank
By cvlover
Posts:  447
Joined:  Thu Apr 23, 2009 4:59 pm
#162107
Matt,
Long time don't see pictures of your CPs in TC. Are you still play with them?
By Frankco
Posts:  14
Joined:  Thu Jan 14, 2010 12:42 am
#162119
Hi Matt,

Yep, it's stuck. I'll let it go and see if they finally upload.
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By Matt
Location: 
Posts:  21111
Joined:  Mon Apr 21, 2008 11:28 pm
#162155
cvlover wrote:Long time don't see pictures of your CPs in TC. Are you still play with them?
I haven't had much time for playing lately. I have many dozens of Venus fly trap cultivars in TC right now and that's about it. They keep me more than sufficiently busy ;)

I do have a couple species of Heliamphora and some Cephalotus...but they may be contaminated. I have to replate and find out :)
By BigBella
Location: 
Posts:  280
Joined:  Fri Jun 17, 2011 9:18 pm
#162163
Frankco wrote: With side by side tests the germination rate with 8% PPM + 1/4th MS for 2 hours was considerably lower than NaDCC. Maybe this is not an optimal amount of PPM and the amount of time was not optimal. So, take that with a grain salt. Also I was testing very clean, lowland seeds. The table may turn toward PPM with other types of seeds and less clean seeds. I’m not sure. Bigbella has very good success with it and I’m sure has worked with more types seeds than I have.
Frank
Frank,
I now generally recommend an 8% PPM solution with 1:2 MS, for four to six hours -- upwards of twelve to twenty-four for lowland Nepenthes and some Heliamphora, whose seed -- native to marshier environments -- naturally harbor far more fungal spores than do any of their highland counterparts . . .
BigBella liked this
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