Post by meizzwang on May 21, 2014 12:32:17 GMT -5
Dorekofu,
while there are other equally as effective methods, I've actually had good success getting sarracenia seeds in vitro using an orchid method called "the green pod method." Let me be frank: it's very difficult, and takes a lot of practice! Other methods might actually be easier, but once you get this method down, it's as easy and satisfying as eating a bowl of ice cream!
All of the following steps are done under a flow hood. Essentially, you take the entire Sarracenia fruit when it's still green (but the seeds are brown and ripe inside) and you soak it in 100% bleach (in other words, you don't dilute the bleach, it typically comes as 5.75% sodium hypochlorite) for 20 minutes. You'll want to swirl it around a lot.
After the 20 minute soak with profuse swirling, you take the entire green pod and dip it in 90% ethanol (90% isopropyl alcohol will work too) and light it on fire! Be very careful and follow all laboratory safety procedures! This will dry up the pod and it dehydrates the nooks and crannies of the pod, thereby sucking out anything that might be hiding there.
The seeds inside that green pod are aseptic, so long as there weren't any holes, cracks, or insects that got into it during development or during the "cleaning" process. Here's the catch: your green pod has to be somewhat clean to begin with in order for this to work.
Now that you have a dry, aseptic pod, place the pod in a sterile petri dish, and cut out a square with a scalpel. Stab the "square" in the middle with the scalpel and twist to open the pod. Ta-dA! you have clean seeds ready to plate.
Main catch about this method: now, you have to keep these flasks in the fridge and let them stratify before placing them under lights to germinate. The lag time increases the chances of contamination, and the hot to cold, cold to hot temperature swing will causes water to condense, which can cause contamination issues. However, if you're careful and keep the surroundings semi-sterile, it shouldn't be an issue whatsoever.
while there are other equally as effective methods, I've actually had good success getting sarracenia seeds in vitro using an orchid method called "the green pod method." Let me be frank: it's very difficult, and takes a lot of practice! Other methods might actually be easier, but once you get this method down, it's as easy and satisfying as eating a bowl of ice cream!
All of the following steps are done under a flow hood. Essentially, you take the entire Sarracenia fruit when it's still green (but the seeds are brown and ripe inside) and you soak it in 100% bleach (in other words, you don't dilute the bleach, it typically comes as 5.75% sodium hypochlorite) for 20 minutes. You'll want to swirl it around a lot.
After the 20 minute soak with profuse swirling, you take the entire green pod and dip it in 90% ethanol (90% isopropyl alcohol will work too) and light it on fire! Be very careful and follow all laboratory safety procedures! This will dry up the pod and it dehydrates the nooks and crannies of the pod, thereby sucking out anything that might be hiding there.
The seeds inside that green pod are aseptic, so long as there weren't any holes, cracks, or insects that got into it during development or during the "cleaning" process. Here's the catch: your green pod has to be somewhat clean to begin with in order for this to work.
Now that you have a dry, aseptic pod, place the pod in a sterile petri dish, and cut out a square with a scalpel. Stab the "square" in the middle with the scalpel and twist to open the pod. Ta-dA! you have clean seeds ready to plate.
Main catch about this method: now, you have to keep these flasks in the fridge and let them stratify before placing them under lights to germinate. The lag time increases the chances of contamination, and the hot to cold, cold to hot temperature swing will causes water to condense, which can cause contamination issues. However, if you're careful and keep the surroundings semi-sterile, it shouldn't be an issue whatsoever.