# fool proof green pod sowing protocol



## TyroneGenade (Jun 30, 2011)

Hi all,

I have a pod of Paph thaianum which is ready for sowing. I want to sow it now while I have chance using the green pod method. I have tried sowing green pod before with dismal success rates. I need a fool proof sterilization protocol for the pod and any advice on seed distribution into the flasks would be much appreciated!

Lastly, has anyone had success sowing on to 1/4 strength MS media (i.e. 1/4 strength N, everything else at normal values). Otherwise, anyone sown on full strength Knudson C with success? These are the flasks I have available and don't really have time to make fresh medium.

Thanks


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## Brian Monk (Jul 3, 2011)

Sterilizing green pods is relatively easy. yo can soak them in pure bleach or CaHypo at saturation. Be aggressive when scrubbing them, and let them soak both before and after scrubbing. Add some kind of soap to your CaHypo solution. I have read the old AOS articles and several studies that show that seeds germinate better with CaHypo than NaHypo, probably because of the sodium. So I would reconsider rinsing the pod after its second soak in CaHypo, and just cut it open. The fewer steps you have, the less likely contamination becomes.

I am also interested in the best media to sow Paph seeds in, but few people are willing to discuss their findings or experiences. I do know that fructose is a better CHO sorce for Paphs than any other saccharide (mon or di).


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## John Boy (Jul 3, 2011)

Hiya Tyrone,
when it comes to sowing Paphiopedilums there’s always that special vibe in the air about finding a good medium for sowing. I have given up on a cheap and working solution, and therefore buy the fancy stuff from Quick Tissue Plant Lab. in the U.K. It’s 100% the best I’ve come across, it covers all Sections as far as my knowledge goes, and instead of risking a good, potentially rare pod: I rather fork out the money.
Sterilisation of green pods:
There’s more than one way to do it. Larger pods I just soak in alcohol for a while and then burn off what’s left after taking the pod out of the alcohol flask. With Restrepia pods you can’t quite do that, because of the obvious “frying of the entire pod issues”. I’d recommend to just stick to alcohol and scrubbing (no fire), which does the trick as well. With a Brachypetalum size green pod I’d do a mixture of both. Given the fact that Brachy pods are hairy and therefore provide a huge surface I initially soak the pod in bleach and tween for an hour, scrubbing it once or twice within the process. After that I hand over to alcohol and flames, paying attention to keep the actual burning time down as much as I can. I burn the pod, trough it back into the alcohol (to cool it off) and burn it again, to be dipped into sterile water as a last step. I’ve been using this method for a while now, and it does work, as long as you keep an eye on the timing when burning small pods. I have a few photos of a green pod sowing, though it’s a larger pod. Enjoy!


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## John Boy (Jul 3, 2011)

Just for you to know what I call a happy Brachypetalum flask, produced with the best Medium I have seen so far....... Anything else I have tried results in plants a fraction of the (leaf) size, with near enough no roots.


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## Bob in Albany N.Y. (Jul 3, 2011)

John Boy, Those are the largest bad ass roots that I've ever seen in a brachy flask. Those seedlings look to have a pretty good leaf span as well. What brachy is in there and how long has it been in the flask? Those are the kind of flasks that I'd like to get.


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## John Boy (Jul 3, 2011)

Hey Bob, that Brachy is what’s been coming to be called or known “a half-album” leucochilum. I’ve had a very fine albino, and it’s been quite a few years ago that I found a good-growing leucochilum in England that I used as a father for it. That plant has near enough no pigmentation in its’ leafs, and its’ flowers show the same trait, going along with a larger than normal flower. So you could say I was playing with “half-album-breeding” before there was a word for it. Actually you can see in that flask that the plant on the right looks like an albino, compared to the other 2 plants. 
Age: I admit that I do allow for more time to produce seedlings like that. The idea behind it is as simple, as it is efficient: I have no plants dying on me (after deflasking), give or take the odd one for the fungus-gods, which at that size rarely happens. The price I’m happily paying is another 3-4 extra months on the flask shelf. No point buying expensive media, rushing magic and loosing plants at the end of the story I thought. I’d say these flasks must have been 14-16 months after sowing. At that stage, and provided you’re able to deflask in the spring: these seedling normally flower about 12-14 months later, which does line them up with the normal leucochilum flowering season in the northern hemisphere. So, looking at things from the other angle: I’m winning a year or two, by allowing more time on the flasking shelf. Let me see if I can dig up another sanderianum picture, though these buggers start the monster rooting only after I take them out of the flasks…


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## John Boy (Jul 3, 2011)

*found it!!!*

My first sanderianum sowing, long....long ago (when I was young), and so proud of these flasks....*that it had to be a white velvet cloth!...(*I wouldn't do that sort of thing these days, or maybe I would, _for an albino sanderinaum sowing_).





Obviously not as monster-like rooted as the Brachy you've seen, but (I'm told) sanderianums never root well in vitro, nothesless: I kept these anchient sanderianums in vitro for about 2 years... (longest ever!).


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## Brian Monk (Jul 3, 2011)

JB - 

Do you sow your seeds directly onto the full-strength TQPL media? I have re-plated several older flasks that were on the TQPL media, that seemed to do well on another media after the reflask.


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## NYEric (Jul 3, 2011)

Great and informative thread. I need to check and find one on pollinating plants.


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## Pete (Jul 3, 2011)

id do something along the lines of what John is suggesting. I would not say you need a pure bleach solution. that seems a bit excessive. i usually scrub with a (clean) toothbrush and alcohol, soak in 10% bleach solution with a drop or two of tween-20, then alcohol and quick flaming before opening.
i have no experience with the tissue quick plant labs media but it seems to look pretty good! i have had excellent experience using sigma P6668 orchid maintenance media or phytotech P668 orchid maintenance media for a variety of genera. for germination (and paph replates) i cut the solution in half. also i supplement with banana.. on the rate of like 10 grams or so per liter.


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## TyroneGenade (Jul 4, 2011)

Hi all,

Thanks for the replies but they came a little late... I had to flask the seed on Saturday.

I just got some generic store bought bleach, add a few drops of tween and soaked the pod for a few minutes and then scrubbed it with a tooth brush and then let is soak for another 10 minute before going to the scrubbed out tissue culture hood and proceeding to next step.

I use 6% H2O2 as a surface sterilant so I gave the pod a squirt with it directly after taking it out the bleach in the hood. It fizzed beautifully as the chlor oxidized the H2O2. Then it got a autoclaved H2O rinse and that was that... Next I simply sliced the pot in a sterile glass petri dish. The pod almost exploded on being cut. The seed was still bone dry (hooray!) and tapped as much as I could into the flasks.

For flasks I'm using plastic tubs which have been sterilized. These are working well. They are tough plastic with snap fast lids. I've had some Chinese Cymbidium seedlings in these now for several months. No problems.

This was a rush job so I had to use my left-over Disa germination mediums which is 1/4 Nitrogen 1/2 strength MS medium with some sugar and banana. I hope it works. So far so good: no contamination. Previous attempts at green pod sowing were already a disaster 48 hours later so I'm feel very optimistic.

I found Phytamax 6668 with banana powder to be excellent for the Paphs as replate medium. I wish I still had some. My Paph insigne x (Knock Knock x Red Glory) seedlings are developing very slowly in the Knudson C.

P.S. I use plastic tubs similar to those seen at http://www.tissuequickplantlabs.com/. I had previously had them steam sterilized at the adjacent hospital but as of late I have simply sprayed the tubs (inside and out) with 6% H2O2, snapped them closed and left them in the sun for a few days before pouring media. Works well so far. The polypropylene allows O2 and CO2 to diffuse across it so venting isn't needed.


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## John Boy (Jul 4, 2011)

Brian Monk said:


> JB -
> 
> Do you sow your seeds directly onto the full-strength TQPL media? I have re-plated several older flasks that were on the TQPL media, that seemed to do well on another media after the reflask.



Morning Brian!
Yes, I do.
I have tried for years to find ways around the rather costly starter-media that Allen provides. Over the years I’ve been in contact with him a lot, not so much as to get his recipe from him, but to find out why his media work as well as they do, or put it this way: *to find out why noting else works. *Besides: his knowledge has been invaluable, and it comes free.

I’ve come to appreciate this contact very much, and these days I don’t mind it the least to pay him for his media. They make all the difference in the world, and because of their complexity it would be madness to even try fiddling with them… even if I had a (biochemical) idea about what I’d be doing, which in any case I don’t…
So, the answer is yes, *I do use them at full strength as sowing and replate medium*. I’ve come to understand the “pointlessness” of messing with Allens’ media, since he knows what he is doing. Over the years, having done my share of trying to cut corners it’s much more important to me to produce the best possible seedlings than to save a few quid on rushing things or using inferior techniques. I guess its okay to say that I’m some sort of a perfectionist when it comes to sowing, and that might also be the reason why I’m allowing for more shelf-time, knowing the ultimate outcome is a better, faster seedling.


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## Brian Monk (Jul 4, 2011)

Using what works makes good sense. I just wish I didn't have to pay for shipping!!!


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## TyroneGenade (Jul 11, 2011)

Well, 9 days later and no contamination. So far so good. Now the wait for germination and then hopefully scores of little Paph thaianums.


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