|
Post by insectivore on Dec 16, 2006 16:03:23 GMT -5
Hey Lloyd, What do you plan on putting in-vitro? Thanks! Cya
|
|
|
Post by lloyd on Dec 16, 2006 19:51:38 GMT -5
I want to put some cephalotus non-carnivorous leaves in. Maybe some of the little Mexican Pings too.
|
|
|
Post by insectivore on Dec 16, 2006 21:09:37 GMT -5
Cool! Let me know how the pings turn out! Cya
|
|
|
Post by lloyd on Dec 25, 2006 13:38:14 GMT -5
Well the house was quiet but I was stirring this morning (literally). I set up 4 jars of cephalotus explants today. I hope the Xmas spirit will keep the micro-organisms away. I realize I need a longer forceps as I kept burning my fingers after I sterilized the forceps with the alcohol lamp. I also am getting a low-end pH meter for Xmas. I'll send some pictures if I have any success.
|
|
|
Post by vraev on Dec 25, 2006 18:31:38 GMT -5
good to know lloyd. Hope they turn out well. Would be nice to have someone make CP's in toronto.
|
|
|
Post by lloyd on Dec 29, 2006 20:42:25 GMT -5
Most of my explants seem to have serious phenolic bleeding. Any suggestions? I'm considering putting them on new medium. I also read that citrate (0.125 w/v%) pre-treatment seems to prevent phenolic bleeding.
|
|
|
Post by tom on Dec 30, 2006 9:14:16 GMT -5
if your flask is large enough, just 'replant' it elsewhere in your flask, where you dont see that blackening in the media. Using charcoal in your media help to attenuate the effects of phenols on explants , if i remember well.
I never used citrate, or read about it unfortunatly.
|
|
|
Post by lloyd on Dec 30, 2006 14:42:05 GMT -5
|
|
|
Post by lloyd on Jan 19, 2007 22:42:29 GMT -5
Well I think all my ceph explants have died probably from phenolic bleeding. Tom found a reference that suggested complete darkness for 2 months to avoid oxidization of the phenolics which evidently leads to toxic substances (free radicals?). I'm going to use a really gentle decontamination next time with a PPM rinse, maybe potassium citrate:citrate as an anti-oxidant and initial darkness. Tomorrow I'm going to start some Sarr and darlingtonia seeds in TC.
Jan. 21: I noticed one of the explants that had turned partly red (stress->anthocyanins?) has turned green. I hope this is a good sign.
|
|
|
Post by lloyd on Feb 3, 2007 20:36:54 GMT -5
I know this is old hat to you guys but it's exciting to me (I lead a quiet life.). I just noticed some roots coming out of my Darlingtonia and S. purp seeds after 2 weeks. One purp seed looked like it might have had a bit of mold (sticking above the medium) a week ago but I took it out and so far no sign of contamination. I'm keeping my fingers crossed.
|
|
|
Post by insectivore on Feb 3, 2007 22:32:33 GMT -5
Good to hear that its starting to work out for ya man! Cya
|
|
|
Post by lloyd on Feb 3, 2007 22:55:31 GMT -5
Thanks.
|
|
|
Post by tom on Feb 3, 2007 23:10:49 GMT -5
great lloyd! I found that some contaminants aren't harmful to the explants, competing for sugar into the medium without hurting the tissue. In the case of valuable flask, keep them anyway, especially in later stages, so you can perhaps rescue them anyway... i did it a few time with Phalaenopsis stemprop... keep us posted on your success
|
|
|
Post by lloyd on Feb 15, 2007 11:06:24 GMT -5
I'm going to make an order to Phytotech. Right now I use agar I bought from Bulkfoods.com which seems Ok to me. It's slightly yellow when dry but as a gel it looks like standard agar, clear and slightly milky. Does anybody think the TC grade agar from Phytotech is worth the extra money? It's a lot more expensive. Here's a useful Canadian chemical supplier: www.jdphotochem.com/They sell small amounts fairly cheaply and the S&H is reasonable, too.
|
|
|
Post by sdeering on Feb 15, 2007 21:56:56 GMT -5
I just looked in a TC book of mine. Through out it always talks about how medias are designed and how different factors affect how it all comes together, works. If other stuff is added it will affect how plants use the nutrients. Eg. regular water instead of distilled. Regular has all sorts of minerals and stuff in it that will affect how the plant uptakes the nutrients its suppose to. Agar can have a lot of undesirable minerals and salts in it if it is not purified. How to tell I don’t know. The book does talk about washing the agar but I think it would be a lot of trouble.
The book even states as to chemical clean jars first because of minerals leaching out of the glass. Also states to clean and rinse containers with distilled water to remove any stuff left behind. Needless to say I have reasonably good luck washing in regular water and drip drying (I think).
If you have a conductivity meter you may be able to mix a test batch of distilled water and some agar and test before and after. I’m not sure if pure agar affects conductivity or not. I looked at some agar on Ebay and it stated it had iodine and sodium in it. This would not be good for tissue culture. Stephen
|
|