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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Cactus tissue culture. * 1
    #1932583 - 09/19/03 01:32 PM (20 years, 4 months ago)

Here's my first success.

San pedro developing new buds in vitro :laugh:



 


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Offlineneuro
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Re: Cactus tissue culture. [Re: Una]
    #1932590 - 09/19/03 01:33 PM (20 years, 4 months ago)

what's your medium and hormone content and ratio?


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InvisibleUna
controlleddemolition

Registered: 03/01/01
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Re: Cactus tissue culture. [Re: neuro]
    #1932650 - 09/19/03 01:44 PM (20 years, 4 months ago)

I can not reveal the recipe but if you are interested in cactus tissue culture this document will give some very usefull tips.

http://plant-tc.coafes.umn.edu/listserv/2000/log0010/msg00077.html





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Offlineneuro
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Re: Cactus tissue culture. [Re: Una]
    #1932680 - 09/19/03 01:50 PM (20 years, 4 months ago)

heh i already know how to do cultures and such... back in school i had this shitty lab every tuesday afternoon and all we did was make cultures.. i was just curious as to your recipe...but if mum's the word, then mum's the word  :lipsrsealed:


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InvisibleDazedSol
old hand

Registered: 08/01/01
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Re: Cactus tissue culture. [Re: Una]
    #1933189 - 09/19/03 04:47 PM (20 years, 4 months ago)

Very fucken cool Una!!! :laugh: 

Please keep us updated on your progress.......

This is on my (really long)list of things to do(different, harder to acquire cacti though)......

anywho.
Hehe, I was gonna ask the same thing as neuro but figured the request would be shot down :smile: With good reason though,..there is a lot of money to made on successful culture formula's ....plus all the time done experimenting and all.........thats alot to just give away 


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Invisibletheshiftingwalls
Divine state

Registered: 06/18/03
Posts: 4,128
Loc: Residing in thee Universa...
Re: Cactus tissue culture. [Re: DazedSol]
    #1933403 - 09/19/03 06:12 PM (20 years, 4 months ago)

Give us the secrect... Or thats just fucked...


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Invisibletheshiftingwalls
Divine state

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Loc: Residing in thee Universa...
Re: Cactus tissue culture. [Re: theshiftingwalls]
    #1933408 - 09/19/03 06:14 PM (20 years, 4 months ago)

Malt extract or agar?


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Offlineneuro
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Re: Cactus tissue culture. [Re: theshiftingwalls]
    #1933482 - 09/19/03 06:54 PM (20 years, 4 months ago)

LOL! Not quite!


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Offlinefelixhigh
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Re: Cactus tissue culture. [Re: neuro]
    #1933797 - 09/19/03 09:41 PM (20 years, 4 months ago)

hehehehe
yeah, MEA, then you insert the Cube spores into the mix and there you have a shroomocactus psychodelicus. =)

very very cool UNA!
could you guys please give us more good links on cactus tissue culture? should i treat them like any other plant then (regarding substrate)?


Edited by felixhigh (09/20/03 01:28 AM)


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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: Cactus tissue culture. [Re: felixhigh]
    #1935653 - 09/20/03 02:21 PM (20 years, 4 months ago)

Of course the medium is not MEA.... That's for fungal growth not for plant growth (although malt extraxt is sometimes added to tissue culture media as an 'undefined'  growth factor).

The cactus medium is based on Murashige and Skoog (MS) salts, sugar (sucrose), agar and growth regulators.

I've searched for many hours on the internet for info on TC of cactus but there's not much info out there. The link i posted earlier sums up (practically) all the info that is out there.

Whenever i have the time i will go visit the library of the Agricultural University in Wageningen because i'm sure that many scientific articles have been published about the subject that i can get my hands on there.

As for me not wanting to share the exaxt recipe; This is something we might want to take to a commerical scale (sometimes). In fact i studied horticulture for 4 years and i did my final thesis on the development of plant tissue culture protocols :laugh: 


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Offlineneuro
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Re: Cactus tissue culture. [Re: Una]
    #1935877 - 09/20/03 04:03 PM (20 years, 4 months ago)

>>As for me not wanting to share the exaxt recipe; This is something we might want to take to a commerical scale (sometimes). In fact i studied horticulture for 4 years and i did my final thesis on the development of plant tissue culture protocols

Ahh so this is a carefully investigated recipe.. i understand now..


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Invisibletheshiftingwalls
Divine state

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Re: Cactus tissue culture. [Re: neuro]
    #1936033 - 09/20/03 05:04 PM (20 years, 4 months ago)

Told you he used agar and malt extract.


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InvisibleUna
controlleddemolition

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Re: Cactus tissue culture. [Re: theshiftingwalls]
    #1937196 - 09/21/03 03:22 AM (20 years, 4 months ago)

Quote:

Told you he used agar and malt extract.
Quote:



I use malt extract for the growth of fungi, haven't used it for TC plants yet. Malt extract is regarded as complex organic supplement of which the effects are not completely understood.



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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: Cactus tissue culture. [Re: Una]
    #1938225 - 09/21/03 03:29 PM (20 years, 4 months ago)

Here's some more pics.....

more pachanoi



Lophophora developing a new pup



Pereskiopsis rooting stage



oh here's some Salvia divinorum (not a cactus i know)




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Offlineneuro
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Re: Cactus tissue culture. [Re: Una]
    #1938533 - 09/21/03 06:17 PM (20 years, 4 months ago)

looks good una..

have you tried producing a callus then growing shoots or roots off the callus in a separate culture?


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OfflineISH
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Re: Cactus tissue culture. [Re: neuro]
    #1938571 - 09/21/03 06:33 PM (20 years, 4 months ago)

Bad Ass, I'm very interested in seeing your progress.

ISH


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OfflineAgarico
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Re: Cactus tissue culture. [Re: Una]
    #1938577 - 09/21/03 06:36 PM (20 years, 4 months ago)

Is it possible to do this without  aureoles? Just tissue?  :cool:

Cloning Salvia that way looks like everything else than a productive method  would :lol:     


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InvisibleEffedS
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Re: Cactus tissue culture. [Re: Una]
    #1939019 - 09/21/03 09:34 PM (20 years, 4 months ago)

Una, you are a true innovator.  :thumbup:


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Invisiblepsilocybe cubensis
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Registered: 08/09/00
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Re: Cactus tissue culture. *DELETED* [Re: Effed]
    #1939068 - 09/21/03 09:58 PM (20 years, 4 months ago)

Post deleted by psilocybe cubensis


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OfflineOpenminded
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Re: Cactus tissue culture. [Re: psilocybe cubensis]
    #1940698 - 09/22/03 12:51 PM (20 years, 4 months ago)

Very, very nice. I've always been fascinated by this sort of tissue culture. I love the way you can simply slice off a bit of something and it will grow into a whole new plant. Well, not quite that simple in practice...
I love growing things, but feel more at home in the lab than in the garden, so this would be a perfect hobby for me!

Please, do keep us informed about your progress. How long did it take for the new shoots to form?


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InvisibleAnand
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Re: Cactus tissue culture. [Re: Openminded]
    #1943811 - 09/23/03 09:30 AM (20 years, 4 months ago)

Una: will MushMush sell plant Tissue Culture supply's in the future? would be nice.... don't know where to get the MS and other ingredients


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Invisibletheshiftingwalls
Divine state

Registered: 06/18/03
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Re: Cactus tissue culture. [Re: Anand]
    #1946529 - 09/23/03 11:44 PM (20 years, 4 months ago)

He would sell the ingredients to make it possible to culture, get it?
Its all marketing :wink:.



Una you better ship to Canada :wink:! You better!


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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: Cactus tissue culture. [Re: theshiftingwalls]
    #1950405 - 09/25/03 04:39 AM (20 years, 4 months ago)

Embryogenesis from callus is possible with many species so i don't see why it's not possible with cacti. Of course you would need a good protocol (which i don't have yet).

The shoots of the SP took 3 weeks to develop.

We will not be selling plant TC supplies since most of the growth regulators that are needed are toxic (carry the skull icon) and these would require special (expensive) transport.


Anand if you are really serious about this is might help you out with some stuff. Or you might contact :info@labassociates.com at http://www.labassociates.com/



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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: Cactus tissue culture. [Re: Una]
    #1957498 - 09/27/03 03:05 PM (20 years, 4 months ago)

I set up some pieces yesterday of this cristate/monstrosus pachanoi.

Here's my flow cabinet. Jars wiith medium and sterilised water on the right. Glass bead heater and more sterilised water on the right:



Surface sterilisation with bleach (toilet cleaner :smile:)



Rinsing with sterile water.



Notice the bleach damage (white tissue)



The damaged tissue is cut off. And the remaining piece is divided.



These pieces are put on the multiplication/initiation medium.



No i cross my fingers and hope some of the areoles develop new buds.


 


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Anonymous

Re: Cactus tissue culture. [Re: Una]
    #1958488 - 09/27/03 10:31 PM (20 years, 4 months ago)

Very interesting stuff. I was talking to my grandmother the other day about growing mushrooms on agar. Then she starts talking about how my greatgrandfather when he was youger got into orchids. He had a full lab and spent all this money on growing thousands of rare orchids. No hybrids. Just pure strains. Then when Hawai came into the picture the market got flooded with orchids. So since he considered himself an artist he took all his plants and equipment and threw them in a pile and burnet them. I wish I had that stuff. Crazy


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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: Cactus tissue culture. [Re: Una]
    #1990544 - 10/08/03 03:50 PM (20 years, 3 months ago)

Here's an update on the San Pedro experiment.

I transferred the pieces to fresh medium and cut away some of the callus. The new shoots are developing steadily:



The Trichocereus bridgesii is now also showing some activity:



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Anonymous

Re: Cactus tissue culture. [Re: neuro]
    #2027654 - 10/21/03 01:33 AM (20 years, 3 months ago)

Callus yields genetic variation.  Bad idea for maintianing cultivars. Good idea for discovering new ones though. 

Stick with the axillary shoot multiplication for maintaining cultivars, while multiplying them. 

Those look nice Una.

Research is very limited in cacti microprop. What is out there is usually not reproducible, important info isconvieniently left out of the methodology. :wink:

I have some journal studies on the subject of cacti microprop. if you are interested.  Explant selection, callus induction, axillary shoot multiplication, somatic embryogenesis, nutritional studies, some specific species studies.  There really wasn't much out there as of 1996.
 


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Offlineneuro
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Re: Cactus tissue culture. [Re: ]
    #2028100 - 10/21/03 08:11 AM (20 years, 3 months ago)

>>I have some journal studies on the subject of cacti microprop. if you are interested. Explant selection, callus induction, axillary shoot multiplication, somatic embryogenesis, nutritional studies, some specific species studies. There really wasn't much out there as of 1996.

I'd be interested...


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Anonymous

Re: Cactus tissue culture. [Re: neuro]
    #2029409 - 10/21/03 04:25 PM (20 years, 3 months ago)

PM me an addy. Which studies are you interested in?


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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: Cactus tissue culture. [Re: Una]
    #2044680 - 10/26/03 12:37 PM (20 years, 3 months ago)

Lophophora williamsii v. jourdaniana



Trichocereus pachanoi



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InvisibleDazedSol
old hand

Registered: 08/01/01
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Re: Cactus tissue culture. [Re: Una]
    #2044693 - 10/26/03 12:42 PM (20 years, 3 months ago)

Hey Una, What kind of lighting do you have those jars under while they grow?


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Adam


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InvisibleUna
controlleddemolition

Registered: 03/01/01
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Re: Cactus tissue culture. [Re: DazedSol]
    #2046503 - 10/27/03 02:17 AM (20 years, 3 months ago)

They grow under simple fluorescent tubes (33/84 cool white)


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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: Cactus tissue culture. [Re: Una]
    #2056126 - 10/30/03 02:28 AM (20 years, 3 months ago)

Yesterday i cut up the newly formed shoots. They had been growing for 2 months. Each shoot yielded between 2-4 new sections that were set up on fresh medium. This gave a mean multiplication factor of 7.

If further multiplication goes with that same number in one year 7^5 plants could be obtained. Not 7^6 because the last cycle the plants can not be cut up but should be rooted in a different medium. Still this would mean 16807 plants in one year starting with one explant. Now THAT is rapid multiplication :thumbup:

Cytokinin concentration seems to influence number of shoots and number of ribs per shoot (higher -> more)

Here's two pics:







 


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OfflinestonErollEr1
The Psilocybinsolution
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Re: Cactus tissue culture. [Re: Una]
    #2056426 - 10/30/03 06:28 AM (20 years, 3 months ago)

And growth would probably be faster if after first initial growth, change lightning into something more powerfull light HPS/MH?[right?]

Peace..


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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: Cactus tissue culture. [Re: stonErollEr1]
    #2056552 - 10/30/03 08:09 AM (20 years, 3 months ago)

Probably not, the atmosphere within the jars does not allow for much assimilation. The medium contains sucrose to supply the plantlets with energy.

The shoots will also develop in the dark but they will remain white/yellow.


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Offlineneuro
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Re: Cactus tissue culture. [Re: Una]
    #2056583 - 10/30/03 08:27 AM (20 years, 3 months ago)

>>If further multiplication goes with that same number in one year 7^5 plants could be obtained. Not 7^6 because the last cycle the plants can not be cut up but should be rooted in a different medium. Still this would mean 16807 plants in one year starting with one explant. Now THAT is rapid multiplication

WOW!  :oogle:


>>Cytokinin concentration seems to influence number of shoots and number of ribs per shoot (higher -> more)

Rather interesting that it affected number of ribs.



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Offlinefelixhigh
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Re: Cactus tissue culture. [Re: neuro]
    #2056798 - 10/30/03 10:30 AM (20 years, 3 months ago)

una, thats it, you've crossed the barrier between the healthy and the insane, please do society a favour and go to intern yourself on a mental hospital.

you leave me without words. it's a little boring to repeat but:
AMAZING!!!


FH


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OfflinestonErollEr1
The Psilocybinsolution
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Re: Cactus tissue culture. [Re: felixhigh]
    #2059435 - 10/31/03 12:17 AM (20 years, 3 months ago)

Yep..this work is just AMAZING..and very very interesting to follow..
great work UNA..

peace..


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InvisibleUna
controlleddemolition

Registered: 03/01/01
Posts: 970
Re: Cactus tissue culture. [Re: Una]
    #2102072 - 11/13/03 02:14 PM (20 years, 2 months ago)

I just received a beautiful variegated Tr. peruvianus and i couldn't resist setting up some new explants. We'll see what comes out, variegated, yellow or green shoots...

Here's the tip of the plant:



Surface sterilisation of the tip in household bleach on a magnetic stirrer plate. This is followed by three rinses with sterile water.



The tip is cut into sections.



And these are placed on the medium.



Once finished the jars are taped with breathable bandage tape. And yes, those are urine sample jars :laugh:



Here's some Lophophora jourdaniana



And some Lophphora williamsii seedlings



My pictures of the san pedro failed but they are developing new shoots on almost all areoles. Multiplication can be even faster than those 16,000 per year!


 


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OfflineEkstaza
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Re: Cactus tissue culture. [Re: Una]
    #2105249 - 11/14/03 07:23 AM (20 years, 2 months ago)

Stupendous. I love to see people with great intellect do great things. Una, you are a credit to the human race.
Wonderful
Amazing
Terrific
Outstanding


Yea, I guess that's enough


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InvisibleUna
controlleddemolition

Registered: 03/01/01
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Re: Cactus tissue culture. [Re: Una]
    #2107117 - 11/14/03 04:47 PM (20 years, 2 months ago)

Ok here's a good pic of the san pedro. Most all of the areoles are swelling and sprouting new shoots. It's more than a week faster than the initial explants reacted. The plants are adapting to their new environment :smile:



And some lophophora williamsii.




 


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InvisibleUna
controlleddemolition

Registered: 03/01/01
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Re: Cactus tissue culture. [Re: Una]
    #2135231 - 11/25/03 02:40 PM (20 years, 2 months ago)

just a little update:



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OfflinestonErollEr1
The Psilocybinsolution
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Re: Cactus tissue culture. [Re: Una]
    #2135247 - 11/25/03 02:50 PM (20 years, 2 months ago)

Keep em comming Una..this is sooo cool.
Looking forward to more updates..

peace..


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Invisiblemedicinebag
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Re: Cactus tissue culture. [Re: stonErollEr1]
    #2136493 - 11/25/03 11:23 PM (20 years, 2 months ago)

What kind of Base is the culture setting in?


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Offlineneuro
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Re: Cactus tissue culture. [Re: medicinebag]
    #2136592 - 11/26/03 12:09 AM (20 years, 2 months ago)

the explants are on agar with nutrition and hormones


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InvisibleUna
controlleddemolition

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Re: Cactus tissue culture. [Re: Una]
    #2227778 - 01/08/04 08:53 AM (20 years, 25 days ago)

These pic were taken a few weeks ago. The pachanoi is growing very nice in tissue culture.

Group photo



Close-up



Loph. williamsii callus cutlure with spontaneous button formation.



Their habitat :wink:



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InvisibleDazedSol
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Re: Cactus tissue culture. [Re: Una]
    #2229031 - 01/08/04 05:14 PM (20 years, 24 days ago)

Very very nice!!!

Wish i had time to experiment more.........ever try tissue culture of cannabis??


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Adam


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OfflineOpenminded
Dicotyledon

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Re: Cactus tissue culture. [Re: DazedSol]
    #2230450 - 01/09/04 05:46 AM (20 years, 24 days ago)

For anyone that's interested in tissue culture, this page:

http://www.oup.co.uk/pdf/bt/slater/ch02.pdf

Might be a good starting point. It gives the composition of a typical culture medium, briefly describes the roles of the different growth regulators, gives examples of them, etc.

Edit: the ratios etc that they give surely won't be optimised for cacti, but I still think it's a page that is worth reading...


Edited by Openminded (01/09/04 05:50 AM)


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Offlineangryjslice
now with 20%more anger
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Re: Cactus tissue culture. [Re: Una]
    #2267795 - 01/24/04 07:56 AM (20 years, 9 days ago)

bump/question
this thread is amazing

una, do you forsee any problems in transfering these back out of the liquid medium there in into soil?

~JSlice~


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Offlineneuro
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Re: Cactus tissue culture. [Re: angryjslice]
    #2268288 - 01/24/04 01:09 PM (20 years, 9 days ago)

There should be no problems what so ever. In a class i took some time ago, we got these african violets with chimera's on them. We isolated the tissue that was exhibiting the chimera and grew out african violets on the culture then transfered back to soil. No problems. Unfortunately the chimera's didn't last and the violets turned green again after some time.


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OfflineYSr
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Re: Cactus tissue culture. [Re: neuro]
    #2935660 - 07/28/04 08:10 AM (19 years, 6 months ago)

Is it possible to grow Salvia the same way?
What kind of media should I use then?

YSr


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Offlineneuro
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Re: Cactus tissue culture. [Re: YSr]
    #2936577 - 07/28/04 02:15 PM (19 years, 6 months ago)

>>Is it possible to grow Salvia the same way?

Yes.

>>What kind of media should I use then?

Agar as the mediu, and proper phytonutrients and most important hormones. Plant Tissue culture is quite a complex thing compared average horticulture so much so that most colleges devote whole classes usually 101/102 and upper level tissue culture 300.

I suggest getting a book and startting from there and also a good supplier of IAA, GA, and the like. Because it all depends on the ratio of plant hormones as to whether you get roots, shoots, or both, or a callus .


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InvisibleStonehenge
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Re: Cactus tissue culture. [Re: neuro]
    #2936801 - 07/28/04 03:02 PM (19 years, 6 months ago)

I think I'd like to take one of those classes. Another one I'd like to take is photoshop. The photoshop tutorials are just about worthless. I'll check to see if any tissue culture classes are offered in my area but I doubt it.


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InvisibleUna
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Re: Cactus tissue culture. [Re: YSr]
    #2936954 - 07/28/04 03:39 PM (19 years, 6 months ago)

Salvia can be grown in tissue culture quite easily. But.....my medium recipe is a secret, that's the way things are in tissue culture land...No one shares their recipes and it's often a long trial and error process to get some decent growth.

Here are two pics...

This plantlet has just started to root...


These guys are ready to be transplanted to soil.


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OfflineYSr
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Re: Cactus tissue culture. [Re: Una]
    #2939467 - 07/29/04 05:30 AM (19 years, 6 months ago)

But could you please give a light introduction of some ingredients
to start experimenting with?

And please give a short description of what IAA and GA is....

YSr


Edited by YSr (07/29/04 05:32 AM)


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InvisibleUna
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Re: Cactus tissue culture. [Re: YSr]
    #2939491 - 07/29/04 05:56 AM (19 years, 6 months ago)

IAA and GA3 are plant growth regulators. They signal the plant to development in a certain way; shoots, roots, callus etc.

A small introduction to plant tissue culture can be found here.


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OfflineYSr
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Re: Cactus tissue culture. [Re: Una]
    #2939507 - 07/29/04 06:18 AM (19 years, 6 months ago)

Do you have some more links to pages where I can read about diffrent
mediums for plant tissue cultures?

I'm very interested and don't know where to start. Love working with
shroom-cultures om agar and liquid karo.

YSr


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InvisibleUna
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Re: Cactus tissue culture. [Re: YSr]
    #2939550 - 07/29/04 06:56 AM (19 years, 6 months ago)



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Offlinefaslimy
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Re: Cactus tissue culture. [Re: Una]
    #2946673 - 07/30/04 10:28 PM (19 years, 5 months ago)

There is another group of people who decided to keep their findings to themselves, they enslaved humanity for world domination. Why keep everyone in the dark on the best methods for things?


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Re: Cactus tissue culture. [Re: faslimy]
    #2946717 - 07/30/04 10:40 PM (19 years, 5 months ago)

i don't see how una's secret formula compares to enslaving humanity in any way. una has a lot of money, time, research, and hard work invested in his business. he is not obligated to give away what he has worked so hard to find. there is nothing wrong with reaping the benefits of your hard work.

una has pointed you in the right direction. if you are really interested, do some research and follow the path that you have been shown.


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OfflineGr0wer
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Re: Cactus tissue culture. [Re: eric_the_red]
    #2946820 - 07/30/04 11:25 PM (19 years, 5 months ago)

So una this is grown on an agar based gel? Do you make it from scratch or buy and custom mix it?

Do you think the liquid cloning gel would work just as well?


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OfflineHooty
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Re: Cactus tissue culture. [Re: Gr0wer]
    #2946839 - 07/30/04 11:32 PM (19 years, 5 months ago)

I think it's very interesting your work with salvia tissue cultures, but exactly what are the benifits (concidering salvias well document ease of rooting cuttings in nothing but water) of tissue culturing other than just interest?


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Re: Cactus tissue culture. [Re: faslimy]
    #2946884 - 07/30/04 11:48 PM (19 years, 5 months ago)

>>There is another group of people who decided to keep their findings to themselves, they enslaved humanity for world domination. Why keep everyone in the dark on the best methods for things?

That's how it is in the culture industry.


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Offlinefaslimy
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Re: Cactus tissue culture. [Re: neuro]
    #2947233 - 07/31/04 01:25 AM (19 years, 5 months ago)

Yes, that's also how it is in any industry. It sickens me to think that cultivated hyrids and techniques can never be available to hobbyist growers. Who simply cannot afford to have the resources to experiment so thoroughly, because of this thing called profit which they have put on it.

However, thankyou for sharing those pictures.


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OfflineYSr
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Re: Cactus tissue culture. [Re: Una]
    #2955090 - 08/02/04 04:42 AM (19 years, 5 months ago)

Una, do you recomend any good books?

YSr


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InvisibleUna
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Re: Cactus tissue culture. [Re: faslimy]
    #2955136 - 08/02/04 05:08 AM (19 years, 5 months ago)

I've had many requests of people that want the recipe for the tissue culture of Salvia. Still, this is one thing i want to keep for myself.

Anyone that has basic knowledge of plant tissue culture will probably figure the recipe out in a few months trying. It's not a fancy thing with exotic ingredients.

Although i see your point of view (all information should be free), we live in a world where this is not the case.

The whole tissue culture thing was expensive to start with. We spend at least 2000 euro on equipment, chemicals and apparatus before we had our first plantlet in agar. Even then it took almost a year before we had a devent recipe/protocol.
Can you imagine that i don't want someone to just runaway with the recipe just like that?

On top of that i think, but i may be wrong here, that most of the people who ask me about the recipe, no little or nothing about tissue culture and would be better of asking for a good book than asking for the recipe.

YSr here you go:

Experiments in Plant Tissue Culture by John H. Dodds, Lorin W. Roberts


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OfflineMystiq_Shaman
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Re: Cactus tissue culture. [Re: Una]
    #2955145 - 08/02/04 05:14 AM (19 years, 5 months ago)

from what you are saying the information to make such a recipe is availble elsewhere so it is not like you are "hiding" new information but are only protecting your own investment.:)

I found this post very intresting. keep on the good work una


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Re: Cactus tissue culture. [Re: Una]
    #2955389 - 08/02/04 08:20 AM (19 years, 5 months ago)

Una, have you thought about selling pre-mixed bags of agar and hormone? I'd suspect you wouldn't have to label the ratios of hormones. I could debate myself either way on this whether it'd be a beneficial thing/novelty for ameture hobbyists (referring to people knowing little or nothing about phytoculture) but at the same time it could be a decent money maker, if only for a while supposing people still can't culture with pre-made mixes.


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OfflineOpenminded
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Re: Cactus tissue culture. [Re: neuro]
    #2955410 - 08/02/04 08:29 AM (19 years, 5 months ago)

"We will not be selling plant TC supplies since most of the growth regulators that are needed are toxic (carry the skull icon) and these would require special (expensive) transport."

:sad:


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Re: Cactus tissue culture. [Re: Una]
    #2955461 - 08/02/04 08:57 AM (19 years, 5 months ago)

Una, what about this book? Is it a good investment?

http://www.amazon.com/exec/obidos/ASIN/0881923613/minmed-20/002-7680887-6166417

YSr


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Offlineneuro
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Re: Cactus tissue culture. [Re: Openminded]
    #2958048 - 08/02/04 09:04 PM (19 years, 5 months ago)

ahh yeah, i had forgotten about that.


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Re: Cactus tissue culture. [Re: neuro]
    #2959100 - 08/03/04 04:58 AM (19 years, 5 months ago)

Where do I buy the hormones? Somebody has a link, please?
I just need about 1 gram of each.

YSr


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Offlineneuro
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Re: Cactus tissue culture. [Re: YSr]
    #2960813 - 08/03/04 03:30 PM (19 years, 5 months ago)

Chemical supply companies should have them. Fischer, Sigma and the like, i suspect Fischer and Sigma requires some sort of docs but i doubt all of them do depending on what their stock of chems entails.

Phytotech labs should be good

http://www.phytotechlab.com


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OfflineYSr
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Re: Cactus tissue culture. [Re: neuro]
    #2963172 - 08/04/04 04:28 AM (19 years, 5 months ago)

There are a lot of auxins. Which one should I use?
The same about cytokines...

http://www.phytotechlab.com/TABSTYLE/ite...H%2DAUX&Bc=

http://www.phytotechlab.com/TABSTYLE/ite...H%2DCYT&Bc=

YSr


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InvisibleUna
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Re: Cactus tissue culture. [Re: YSr]
    #2963820 - 08/04/04 10:57 AM (19 years, 5 months ago)

Often used cytokinins are BAP and kinetin

Often used auxins are NAA, IAA and IBA. 2,4-D is not often used because it can cause mutations.

The reason why these are used is because all are autoclavable (although some activity might still be lost) and not too costly. It should be noted that IAA loses it's activity in light in about two weeks.

If you don't have a milligram scale i suggest you buy stock solutions (1mg/ml) and use a pipette or syringe for dosage.

As a basal medium i would suggest Murashige&Skoog or Gamborg B5. I use a mixture that contain macro/micro elements and vitamins.

I assume you have a pH meter because the pH of the medium is very important. You can adjust it with 0,1 M NaOH or HCl.

If i have time i will do a more complete write up about the subject, there's just so much worth mentioning.....

Good luck!


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Re: Cactus tissue culture. [Re: Una]
    #2966390 - 08/04/04 11:28 PM (19 years, 5 months ago)

>>2,4-D is not often used because it can cause mutations.

2,4-D is used as an insecticide because those plants that it kills cannot metabolize that auxin like it can other and natural auxins.

>>there's just so much worth mentioning.....

which is why everyone should get themselves a text book and start reading.


edit: uhh i just noticed this after too day... insecticide = herbicide i'm just stupid and wasn't paying attention to what i was writing.


Edited by neuro (08/06/04 05:28 PM)


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OfflineYSr
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Re: Cactus tissue culture. [Re: neuro]
    #2966724 - 08/05/04 02:01 AM (19 years, 5 months ago)

I've ordered two of them from Amazon...

YSr


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Offlinefelixhigh
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Re: Cactus tissue culture. [Re: YSr]
    #2968659 - 08/05/04 03:35 PM (19 years, 5 months ago)

knowledge is never too much, i believe! :wink:


FH


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Re: Cactus tissue culture. [Re: felixhigh]
    #6912794 - 05/14/07 05:09 AM (16 years, 8 months ago)

very interesting, thought it could use a :bump:


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Re: Cactus tissue culture. [Re: Simisu]
    #6913708 - 05/14/07 11:14 AM (16 years, 8 months ago)

nice nice!

i hope to start an equal project in the near future!

where do you get these plant tubes?
http://www.shroomery.org/forums/files/080503-46/72157-pereskiopsis.jpg
...and what are they called?

greetz


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Offlinebuddhahoodlum
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Re: Cactus tissue culture. [Re: Entersandman]
    #6990962 - 05/31/07 11:10 AM (16 years, 7 months ago)

where do you get these plant tubes?
http://www.shroomery.org/forums/files/080503-46/72157-pereskiopsis.jpg
...and what are they called?


Doe's anyone know the answer to this question?


--------------------
“I believe that water will one day be employed as fuel, that hydrogen and oxygen which constitute it, used singly or together, will furnish an inexhaustible source of heat and light, of an intensity of which coal is not capable.” – Jules Verne, T

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Invisiblethallus
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Re: Cactus tissue culture. [Re: buddhahoodlum]
    #6991686 - 05/31/07 02:02 PM (16 years, 7 months ago)

Good read:thumbup:


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OfflineFreeSporePrints
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Re: Cactus tissue culture. [Re: Una]
    #8391525 - 05/12/08 02:27 AM (15 years, 8 months ago)

Hi mates,
i hope you can be interested to give a hand with our micropropagation project:

http://www.shroomery.org/forums/showflat.php?Cat=0&Number=8391485&page=0&vc=1&PHPSESSID=#Post8391485

Bye,
Fabio


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Re: Cactus tissue culture. [Re: FreeSporePrints]
    #8395641 - 05/13/08 07:27 AM (15 years, 8 months ago)

please do not dig up old threads to plug a thread you have already started!!


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Re: Cactus tissue culture. [Re: Una]
    #8395642 - 05/13/08 07:27 AM (15 years, 8 months ago)

i am going to leave this thread open due to good info but if you are going to use it to plug your thread then i will re-lock it.


Edited by royer9864 (05/13/08 07:29 AM)


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OfflineFreeSporePrints
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Re: Cactus tissue culture. [Re: royer]
    #8395783 - 05/13/08 08:41 AM (15 years, 8 months ago)

Hi royer9864,
i've added the link to link this thread with the other project. I believe a lot of people here are interested in the micropropagation project, and i don't earn anything doing this..i simply do this for the community..so gently next time think if a thing made can be useful for the users or not and in this case i believe yes :smile:

Nobody against you,
Fabio


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Re: Cactus tissue culture. [Re: royer]
    #8395849 - 05/13/08 09:08 AM (15 years, 8 months ago)

Quote:

royer9864 said:
i am going to leave this thread open due to good info but if you are going to use it to plug your thread then i will re-lock it.




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Re: Cactus tissue culture. [Re: royer]
    #8437142 - 05/23/08 02:30 PM (15 years, 8 months ago)

Thanks for stopping micropropagation to micropropagate users!

Fabio


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Re: Cactus tissue culture. [Re: FreeSporePrints]
    #8439423 - 05/24/08 04:58 AM (15 years, 8 months ago)

i did not stop anything, have a nice day


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OfflineFreeSporePrints
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Re: Cactus tissue culture. [Re: Una]
    #10433281 - 06/01/09 12:11 AM (14 years, 7 months ago)

Quote:

Una said:
I can not reveal the recipe but if you are interested in cactus tissue culture this document will give some very usefull tips.

http://plant-tc.coafes.umn.edu/listserv/2000/log0010/msg00077.html





Clicking on the link of Una doesn't work actually, but this works!

http://plant-tc.cfans.umn.edu/listserv/2000/log0010/msg00077.html

I quote also the text:

Quote:

    * Subject: cacti recepy
    * From: "R. Wellens" <rwellens@ZEELANDNET.NL>
    * Date: Tue, 10 Oct 2000 19:43:42 +0200
    * Reply-To: Plant Tissue Culture <PLANT-TC@TC.UMN.EDU>
    * Sender: Plant Tissue Culture <PLANT-TC@TC.UMN.EDU>

Dear Diana,

There is no such thing as a cactus recepy; there is a huge diversity within
the cactaceae and almost each different one asks for a different approach.
In general (very general!) most species will grow on a 1/2 MS or 1/2 B5
medium supplemented with BA in concentrations varying from 0.1 to 10 mg/l.
Some cacti are extremely hard to propagate, for instance the cold-hardy ones
from North-America (Sclerocactus, Pediocactus etc.). So are cacti like
Ariocarpus, Aztekium etc, the slow growers from Mexico.
Relatively easy are most (not all!!) of the Rebutia's, Mammillaria's,
Gymnocalycium's etc.
Normally they are multiplied using areole activation. This can be done on
the medium described above and is sometimes enhanced by culture in the dark
for 2-3 weeks.
You can take pieces of cactus with at least 3-4 areoles and cultivate them
in the dark for 3 weeks on the described medium.
When you cannot start from a sterile (seedling) culture and must start with
an adult plant you will find it hard to desinfest. Normally if that happens
we decide to cut off the biggest spines and sterilize only the top meristem.
We use a 2% bleach for a relatively long time (1 hour or more) and shake the
culture very frequently. Sometimes we add an dropp of dish-washingsoap to
this or you can use a drop of Tween. You still might find fungal infections,
even after a long sterilization. But once you have a single clean cactus you
can easily start the multiplicationprocess with areole activation, although
you the hard-to-do ones mentioned above will ask a lot of time and effort to do so.
Rooting is mostly not a big problem, but again, it is a problem for some species.
Rooting for easy species happens on plain 1/2MS without any auxins, but in
individual cases an auxin like IBA might be helpful.
A main problem in producing cati is hyperhydricity or vitrification.
This can be reduced by using vessels or jars that have air-filtering
capacity, not to much because then they dry out.
So once you know what type of cactus you are going to multiply you will have
to develop its own perfect protocol, which can be based on this information.
However, it is not complete, there are a few articles in which specific
cacti were produced with specific techniques.
Callus culture or somatic embryogenesis is so far not studied by us and is
scarcely found in literature with little success.
I hope this will help you, you can also see some literature on different species.
Success with your productions.

Robert Wellens
Succulent Tissue Culture





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OfflineFreeSporePrints
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Re: Cactus tissue culture. [Re: FreeSporePrints]
    #12547466 - 05/12/10 08:22 AM (13 years, 8 months ago)

Nobody tried with tissue culture?

Fabio


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OfflineOzzy_shroomer
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Re: Cactus tissue culture. [Re: FreeSporePrints]
    #12558609 - 05/13/10 09:46 PM (13 years, 8 months ago)

Sorry i hate doing this to old threads but found this a great resouce also have somthing to add and have the PDF if anyone wants it

Inoculation
Explants were cultured in culture bottles containing MS (Murashige and Skoog, 1962) basal media supplemented
with benzyladenine (BA) (0.5, 1.0, 1.5, 3.0 and 5.0 mg L-1) or Kinetin (Kin) (0.5, 1.0, 1.5, 3.0 and 5.0 mg L-1) alone
or in combination with naphthaleneacetic acid (NAA) (0.5 mg L-1). All media were prepared by standard procedures
and the culture bottles were transferred to growth chamber with a photoperiod of 16 h light and 8 h of dark at 25ºC ±
2 ºC and incubated for 3 months


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InvisibleBig L
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Re: Cactus tissue culture. [Re: Ozzy_shroomer]
    #12558903 - 05/13/10 10:30 PM (13 years, 8 months ago)

Nice bump.

I fully support bumping any of Una's threads.
Una is the bees knees.


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OfflinetheMallacht
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Re: Cactus tissue culture. [Re: Big L]
    #12558938 - 05/13/10 10:34 PM (13 years, 8 months ago)

The bees knees INDEED!

This thread is great... I clicked and just started reading the entirety of the OP, was SUPER impressed and immediately wanted to post and tell Una how awesome he is and ask about 10 questions, then I realized that it was a 2 yr old thread.


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InvisibleBig L
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Re: Cactus tissue culture. [Re: theMallacht]
    #12558965 - 05/13/10 10:38 PM (13 years, 8 months ago)

I highly highly recommend viewing Una's "Main threads" list.


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OfflinePsychdelicSpore
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Re: Cactus tissue culture. [Re: Una] * 1
    #28084253 - 12/05/22 05:36 AM (1 year, 1 month ago)

Why post BS and not tell people how to make it? Why bring it up? This is a Forum! That means don't BS people a bring up something that doesn't matter! How about delete this whole post because it doesn't teach anyone 💩! Stop posting crap that has no info on a DIY. You help no one and post crap to make yourself feel better. 😆😂🤣 GTFOH!


Edited by PsychdelicSpore (12/05/22 05:38 AM)


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