# Plant Tissue Culture



## bombbudpuffa (Jul 19, 2011)

Thought i'd share a few pics of one of my hobbies. Its not really that hard to do but theres a lot of info to remember. I took a couple of months to read and learn about it before attempting it. Cleanliness really is next to godliness when it comes to growing plants in a tube though.

I've tried it with cannabis with marginal success. I could get growth but my sterilization technique for canna isnt what it should be. Right now i'm practicing with carnivorous plants. They aren't only some of the easiest species to culture but also some of the funnest. I mean, they eat things.

1. Venus Flytrap started from seed in vitro
2. Venus flytrap seedling in vitro
3. Flytrap flower stalk with lots of growth
4. Flytrap leaf with several divisions
5. Drosera or sundews germinated in vitro
6. Sundews germed in vitro

I'll probably give canna another shot once I perfect my technique a lil.


----------



## Locked (Jul 19, 2011)

Very cool and very interesting.....keep us posted puff.


----------



## 4u2sm0ke (Jul 19, 2011)

pulling up my milk crate and have pen and papper at the read...

:ciao:  give me a learning *Bombbudpuffa*!!!

:48:


----------



## bombbudpuffa (Jul 19, 2011)

HL-Will do

4U-How about I just point you in the right direction?

This guys videos are very detailed and he tells you everything from where to get supplies down to how to do it. I will warn you that every plant needs different hormones to be tissue cultured successfully. There are usually "how tos" called protocols for all of the different plants you can do it with but I can find zero for canna so you'll have to find the right combination and strengths for canna through experimentation.

hxxp://www.youtube.com/user/fbt2007


----------



## ozzydiodude (Jul 19, 2011)




----------



## bombbudpuffa (Jul 19, 2011)

If you want to learn I can throw up some links and recommend some books for you guys.


----------



## ozzydiodude (Jul 19, 2011)

I don't watch vids but throw up them up for others, I'll read everything I can and put in alot of dumb looks(natural for me)


----------



## bombbudpuffa (Jul 19, 2011)

Seriously Ozzy...if you want to learn watching videos would be quicker because theyre going to use terms in the books that you wont understand. The vids will let you actually see what theyre talking about and kind of be like a refresher course. Not to mention, the vids are super boring so you can imagine what the books are like.


----------



## ozzydiodude (Jul 19, 2011)

But I got to buy a new comp to run videos. Right now looking thing I don't understand up is the only thing other the PT that is keeping me sanely insane


----------



## pawpaw (Jul 19, 2011)

Very cool bombbudpuffa, are you using a laminar flow hood?


----------



## bombbudpuffa (Jul 19, 2011)

Supplies-

hxxp://hometissueculture.org/cataloghtcgsupplies.htm#Media%20Zero

Random links for basic understanding-

hxxp://www.bio.davidson.edu/people/kabernd/seminar/2002/method/amy/aj.htm

hxxp://www.scribd.com/doc/3544465/Plant-Tissue-Culture

hxxp://tech.groups.yahoo.com/group/hometissueculture/

hxxp://www.omnisterra.com/botany/cp/slides/tc/tc.htm

hxxp://www.ccebook.org/preview/0881923613/Plants-from-Test-Tubes-An-Introduction-to-Micropropagation

hxxp://csm.jmu.edu/biology/renfromh/pop/pctc/tcstart.htm

hxxp://www.flytrapcare.com/tissue-culture-basics.html

hxxp://www.archive.org/stream/handbookofplantt00whit/handbookofplantt00whit_djvu.txt

That should be enough to keep you busy for a while. Tried to just post the easy to understand links.


----------



## bombbudpuffa (Jul 19, 2011)

pp-nope.


----------



## pawpaw (Jul 19, 2011)

bombbudpuffa said:
			
		

> pp-nope.


 
Wow:holysheep: 

I've got two books:
*Plants from Test Tubes: An Introduction to Micropropagation *[Lydiane Kyte, John Kleyn] 
and
*Introduction to Plant Tissue Culture *[M.K.Razdan]


I have only skimmed these but the first looks quite good as a howto. The second looks even more technical than I want to get.  I wouldvery much like to know which books you recommend.


Thanks.


----------



## bombbudpuffa (Jul 20, 2011)

The first one you listed is the best one imo. Easy to understand with lots of info and even a section on protocols.


----------



## Locked (Jul 20, 2011)

Looks like school is in session....putting on my thinking cap....

Thanks for the links puff. Good to see ya posting again.


----------



## pawpaw (Jul 20, 2011)

I have two pdfs that may be of interest to you.  one is titled,

INFLUENCE OF CULTIVAR, EXPLANT SOURCE AND PLANT GROWTH
REGULATOR ON CALLUS INDUCTION AND PLANT REGENERATION
OF CANNABIS SATIVA L.


> hemp. Hemphil et al. (1978), Fisse et al. (1981), Mac-
> Kinnon et al. (2000) and Feeney and Punja (2003)
> reported that callus readily produced roots but was
> unreceptive to shoot formation. Mandolino and Ranalli
> ...


The other is

Assessment of the Genetic Stability of
Micropropagated Plants of Cannabis
sativa by ISSR Markers


> Inter-simple sequence repeat (ISSR) markers were used to evaluate
> the genetic stability of the micropropagated plants of Cannabis
> sativa over 30 passages in culture and hardening in soil for 8
> months. A total of 15 ISSR primers resulted in 115 distinct and
> ...



I've tried to upload a zip of the first.  It's 8.3 MB zipped. But after clicking on upload the Manage  Attachments dialog displays the please wait msg then evetually closes with no joy.  Can I have a clue? 

View attachment TissueCulture Geneticstability.pdf


View attachment TissueCulture_1.zip


----------



## Mutt (Jul 20, 2011)

Howdy BBP nice to see ya about.


> sterilization technique


What is your lab like that your are doing cultures in?
Sterilization is limited to availability of contaminates. 
Can you describe your environment in your process? 
Got a lot of experience with "contamination" in "another" hobby. might be able to help ya out. 
Are you PCn all of your equipment to at least 15psi for at least 10-15 minutes?


----------



## Locked (Jul 20, 2011)

Mutt said:
			
		

> Got a lot of experience with "contamination" in "another" hobby. might be able to help ya out.



Sounds quite _*Magical*_ Mutt.........


----------



## Mutt (Jul 20, 2011)

Hamster Lewis said:
			
		

> Sounds quite _*Magical*_ Mutt.........


it's not much difference then doing any other kind of culture.


----------



## ozzydiodude (Jul 20, 2011)

That's some of the best "go to sleep " reading I had done in awhile. Very interesting was to keep strains alive in a small space. This might be the way for a person to keep a living sample of the strains that truely need to be saved for future generations


----------



## pawpaw (Jul 20, 2011)

I hope to use it to keep mothers of multiple strains going.  I don't have room to do it otherwise.


----------



## pawpaw (Jul 20, 2011)

and here's a link to the complete book* The Biotechnology of Cannabis sativa*

hXXp://www.scribd.com/doc/14571756/The-Biotechnology-of-Cannabis-Sativa


----------



## Mutt (Jul 20, 2011)

ozzydiodude said:
			
		

> That's some of the best "go to sleep " reading I had done in awhile. Very interesting was to keep strains alive in a small space. This might be the way for a person to keep a living sample of the strains that truely need to be saved for future generations


pre98BBK the most isolated strain ever perfect candidate... Cindy was whored out to the point of "madame" status...she's pimpin others out....might need to keep her before she contracts a MJ venereal disease. :rofl:
I know I have several turns with her  with other strange dudes, a random female, and a brother with a step sister about 
Some sick $#!+ goes on in my garden from time to time when cindy shows up. :48:


----------



## bombbudpuffa (Jul 21, 2011)

pp-Thank you very much. Seems I need to play around with another hormone. I've been using BAP and kinetin. I can get callus with them both but no offshoots. Thanks again!

Mutt-My "lab" is my living room. I make sure everything is cleaned prior to culturing and theres no moving air. I used to use the bathroom but it gets cramped in there. I rarely get contams but seem to always get them with canna. My PCer is only 12psi so I usually leave things in it for 30-40 minutes to compensate.


----------



## pawpaw (Jul 21, 2011)

I got the missing pdf to upload -- all my ...D'oh!

http://www.marijuanapassion.com/forum/showpost.php?p=705885&postcount=16


----------

