neilshieh

I used a broken 10 gallon tank I had as a clean box and got a cheap mister and basically sprayed everything I could with 70% rubbing alcohol. One thing i wasn't too sure about was when I kept the scissors and tweezers in the rubbing alcohol I wasn't sure if I was supposed to rinse it off with sterile water in between uses. :/ hopefully the alcohol with the culture can only help haha. we will see. no signs of mold or fungus yet, but then again it may take some time to develop.

neilshieh

here's the lab handout provided by Dr. Carol Stiff over at http://www.hometissueculture.org/

her information has been immensely helpful and a great place to get supplies at!

https://www.dropbox.com/s/zt3q4xh3ye...utJune2012.pdf

she has given me many more documents regarding tissue culture and aquatic plants, i will upload those upon request. all credit goes to Dr. Carol Stiff

neilshieh

update: in hindsight i probably should have used some kind of hormone because the initial roots all died and turn brown. I see lots of steady new growth and roots though! but this is going at a ridiculously slow rate.... oh well it was my first time haha. i will post pictures when its more grown in. No fungus in any of the containers! but i had it easy since i was using in vitro plants haha

ferchu22

Great! I guess fungus is the main problem in TC. If you can control that, it's just a matter of time and patience to get the right protocol/procedure for your pant.
Best regards,



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AaronT

I haven't done it myself, but I do believe the hormones are key. You don't really need roots until you're ready to take them out of culture. I believe Ghazanfar used the hormone that triggers the plant to multiply and then when he was on the last transplant he would also add rooting hormone.

neilshieh

yeah I have BPA for the growth and IBA for the roots except since these were grown in vitro already using more hormones isn't really necessary and at that point we're doing subculture. But I think the plant didn't take the shipping from Italy too well so perhaps i should have put in cytokins to compensate for the plant die off.
I will be sure to use hormones when I decide to start round two. Right now they're just growing like plants in regular substrate.

Should i make a batch with BPA and transplant some of the plants? any thoughts guys?

AaronT

Don't fix it if it's not broken. Just wait until you transplant and use the hormones then.

neilshieh

yeah that was what I was thinking too. Slow and steady then :P

I wonder if there are any cons to using hormone stimulated fast growth. I mean usually with animals that produces negative side effects like weak bones etc, are there any for plants? I mean if hormones don't produce a negative effect why don't we just use them with all plants?

neilshieh

Here are some quick iphone pictures! You can see all the die off but you can also see the new growth + root mass.

ferchu22

Nice pics!
I remember, as Aaron said, that Ghazanfar uses an hormone first to trigger shoot growth, then you have to move the explant to a different media to multiply shoots (up to you have as many as you whant), and finnally move those to last media, with another hormone to trigger roots growth. Check Ghazanfar blog for detailed info.
Best regards,

Fernando

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