Micropropigation using Plant Tissue Culture

[ganjatarian]

Rockin Good Kansasgrown. I can't wait to see your pics and your encouragment exudes through your posts. I have tried to be EXTREEEEMLY steril. Completely bleached and then alcohol sprayed the bathroom im using as a lab.

Myst- you have to do a 10 to 1 bleach wash for 10 mintes....then follow with a 70% alcohol wash....then 3 rinses with steril water before cutting off the bleached/ dead ends and then placing them into the agar.

follow up:
I may have added too much water to my pressure canner, I have a thin layer of liquid slidding around the top of my hardened agar medium. Talked to a guy who said 6 cups of water was enough for a 20 minute canning session and wont drown or spill over into the baby jars or more importantly.....drying the canner out.

Has been almost a week and the small leaf (codeldon) look too bleached out to make it and have sunk into the agar. Some samples look like I may not have cut enough bleached tip off and they are browning. I do not see any growth of fungus or vegitation yet but still got my fingers crossed and will start new batch soon.

Thanks G

 

[MIZZ ELVIS]

Ganjatarian, You should try sampling plant material from different areas of the plant.....I suggest scraping down a stem to the cambium using a sterile razorblade(almost as you would to increase the rooting area for a cutting), and then give those tissues a run. Also, I'm surprised the cotyledon didn't work out, Although I've never personally heard of anyone using embryonic leaf tissue for culture, Im sure you can get it to work. Good luck and I'll be around to see how your progressing....Godspeed.

 

[guerilla family]

This is very interesting! I will be keeping an eye on this thread. Thanks for sharing, everyone!

GFam:hi:

 

[kansasgrown]

Mold and fungus are your 2 main enemies. If you do not have a true hepa filter flow hood you might want to get or make a clean box and work from there. If your funds are real low then take a shower, dress yourself in freshly cleaned clothes, really clean your bathroom and make sure that all ventilation is off before starting the process.

 

[kansasgrown]

Looks like if I will not be able to get the pictures of my room for an additional week. I will have to fly my brother out to tend to the garden. I fell and heard a very loud pop in my ankle...and it is now the size of a small melon......time for some more vicadin....and a trip to the local emergency room...think I may have broken it. :sweating

 

[ganjatarian]

Kansasgrown.... I realy hope that you stay off that ankle and get some rest. Don't worry about pics. take good care of yourself first.

Mizz Elviz. I love that idea and will add it to my experiment for next time. I tried to take different samples from different areas of different plants. I even popped 3 seeds in a jar to see if they'll sprout. I did a lemon tree and am seeing little white something on the tip.....could it be a sprout? Oh and thanks for the proper spelling of cotyledon. I was too lazy to look it up and Thanks for the suggestions.

To be continued....

 

[ganjatarian]

I ordered more baby food jars, fresh hormones and a membership to kitchencultureeducation.org and I got a free DVD produced by Carol M. Stiff.
her technique varries from Bill Grahams Super starts DVD.

Carol recommends never using Denatured alcohol, she only mix'smurashige& skoog basal medium w/ vitamins and BAP (benzylaminopurine), and PPM (plant preservative mixture)to her base agar/ sugar base... she does not add NAA (Naphthalene Acetic Acid) in the first phase and she also washs her plant material using Alcohol wash then bleach wash and then steril water. Also, she said with some plants you don't need agar and can use cotton balls. She also recommends using food coloring in your medium to diferentiate the hormones being used. I will try that in my second phase solution so I don't get cultures mixed up. she also stores her jars in a sealable bag so bacteria doesn't creep into the jars as they expand and contract.

This A.M I mixed another batch of 25 using Bill Grahams method and this time I will only take samples of MJ nodes and axillary buds. I also hope to get a new camera this week to start showing the results of my first attempt.

In my humble oppinion I felt much more confidant doing PTC after watching Bill Grahams DVD so I would recommend that for starting out but its nice to watch Carols DVD and see the variations and she also takes you to a huge lab/ nursery in FL that makes your jaw drop at the level this has been taken to. I would love to a nursery like that full of cannabis..... Maybe someday soon.

G

 

[ganjatarian]

its been a week + since my first try and most of my cultures have had bacteria. Im down to about 10 left out of 27 tissue cultures I took. Four of them are from a lemon tree and 2 are showing white sprouts:cool0010: but no roots or calus. Fingers crossed.

I also ordered different hormone, agar and murashige basal medium. If you know what your doing you can put together your own kit alot cheeper than what I bought mine for. Here is the link.
http://www.phytotechlab.com/

came across another article from Growing Edge Magazin.
http://www.growingedge.com/magazine/back_issues/view_article.php3?AID=190326

 

[makala]

what hormones are working for you? a simple answer for a simple mind would be great.

 

[canndo]

NAA, TDZ and GA3 are, in my opinion the best. I have tried quite a few others. This is for establishment and multiplication. You would do well to use a biocide (like PPM). In fact I don't need to use a hood at all when I use enough - in the 3 ml per liter range. I dont yet know how to post pictures here - I've been doing cannibis micropropagation for over a year now (well, much longer but I didn't have the correct hormones down).

 

[primeform]

Has anyone been doing this recently? I read something about bringing back the potency and vigor in strains by using this method. Anyone got some good info on this?

 

[ttystikk]

I keep hearing about how a strains' vigor diminishes the more you clone it, but even after years and dozens of repetitions I never saw it. Thus, if cloning is simpler, it stands to reason that for most of us it would be the logical choice. That said, not only is this just peggin' the geek meter for me, but it might be a killer app for sea of green methods, or for those wishing to spread their home-bred genetics far n wide. This could be the ultimate answer to a guerrilla gardener's prayers, right?

 

[primeform]

dunno i just want all the strains to be back at there PEEK vigor and potency. If this can do that its well worth looking into.

 

[Relaxed420]

Just started looking into this for a larger garden, maybe even doing a clone thing with this idea. How does cost compare between this and cloning traditionally for example sell these before they go into the final resting spot so growera buying can go directly into medium they like to use..

 

[swisscheese]

Any updates from anyone? I am building a hood with a hepa filter and clean room just to do this. I have heard about phytotechlab.com and super starts any reviews around the farm? I am looking for anyone's experience with tissue culture. Thanks.

 

[western5]

marijuana from test tubes is now available on kindle through amazon

 

[Sunstate]

Good morning everyone,
I’m planning of using maristem cloning for propagation. Does anyone know about this type of cloning? Appreciate any help.