orchid seeds in vitro germination

[skyhook]

 

 

 

 

 

 

 

 

 

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This is quite a big pod of an orchid. I've just flasked it, and I took a few pictures. Orchids seeds can be germinated in bottles, and it was first done probably a century ago by someone. This pod is still green. If it is too immature, none will germinate. Green pods just before it turns yellow and split seems to be the preferred stage to flask orchids. I do not know what stage this pod is at, anyway I go ahead and flask it.

I got some jam bottles, clean them. I cooked some agar nutrient, using a recipe I got off a website. fill a bit of the agar nutrient into the jam bottles. I'm not using the commercial plant nutrient, as I can't find it at nurseries yet and I didn't order online. The bottles are steamed 3 times at 12 hours interval for about 30 minutes. Supposed to be 24hrs interval, but I'm impatient. If I have a pressure steaming device , I can steam it just one time. This time ,I steamed those bottles in an old rice cooker, which is not used after we bought a new one. Not a good idea actually. but that's what I did.

After 2-3 days of preparations, I put the bottles in a new fish tank, and cover with a yellow trash bag. I prepare a weak bleach solution in a spray bottle in 1:20 concentration, sprayed everything in the interior of the fish tank with it, and also into the trash bag which is unused before. This is to kill the germs in the air inside the tank. The trash bag covered the fish tank perfectly. I tape up the edges. Left this for at least 20minutes. oh, and the pod is soak in a bottle of bleach and detergent for 20 minutes too. as well as the tweezer.

After 20 minutes or more, time for some action. I cut open the pod, and transferred the seeds into the several bottles. I could only do up 8 bottles, there are a lot more seeds left over. I disposed them. quite a waste. This is handled with the trash bag as an isolating layer. The bottles should be quickily opened, transfer seeds and closed.

After that, I can safely open up the trash bag, remove the bottles and I washed it under tap to remove the bleach smell.

I leave it for 2 days on the table. To my dismay, 6 of 8 bottles were contaminated as a milky liquid or some stuff start to appear. LOL... only left 2 bottles, which I presume is the first 2 bottles I did. The rest of the bottles are quickily discarded. Now is probably the third day as I'm typing this. I've done this for probably 10 times , done some dendrobiums, oncidiums, and spathoglotis. I get them to the 2 months stage, after that they deteriorate as they need to reflask.

The risk of this is the effect of bleach on human health, which I'm quite worried, but I just hope I don't get problems in future. and the next is unknown pathogens which slips into the flask. There are things which I could have done, I'll try to do next time.

 

anyone can comment on this if you want..

Edited August 9, 2010 by skyhook

[skyhook]

         

 

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It has been several months. The seeds grew into seedlings. I think the pros might be able to get their plantlets this size at a much shorter time. I'm satisfied that I get them to this stage.

The flasks are not vented. It is said that ethylene build up inside and the seedlings will be affected. I did several flasks. I used the charcoal tablets for most flask, and have two flasks without using charcoal tablet. The ones with charcoal grew better in comparison. Probably the charcoal absorbed some stuff, and there could be some substances in charcoal that promote better growth, phen* something. But the seeds to pinhead-size protocorms stage can probably use less or no charcoal.

 

sorry about the lousy pics.

 

I guess the next thing I can do is to vent the flasks, haven't figure that out yet. Cotton plugs and aluminum sheets. The last time I tried, the aluminum sheet covered bottles all got contaminated.

Edited December 17, 2010 by skyhook