Marijuana Tissue Culture Success!

A

angrygrower

Member
im going through the post now


  • tupperware container on its side... tools are alcohol flamed​




Alcohol is not going to kill pathogens. Even with a flame, you would have to keep it there for over 60 seconds. i dont have the article that demonstrated how ineffective it is. you would likely have high rates of contamination. there's a reason why ceramic/glass bead sterilizers are used in the industry. i cant imagine how the tupperware on it's side would do anything but block the airflow, and increase contamination


  • Correct, I grow callus' of a few dozen strains, ..... this does not replace cloning...​




if you're growing from callus culture, you're likely to get genetic variability. this is why callus culture isn't the industry standard.

page4

  • contamination issues happen because the explant(node,leaf,flower,seed,etc) has either been A(sterilized so much that it retains useless dna, does not flourish, and/or B) has not been sterilized enough to remove unwanted materials​





It's contaminated because you over sterilized it?? really?


  • I HAVE NEVER INSERTED A BIOLUMINESCENCE GENE INTO marijuana.
    ​




this whole talk about bioluminescence in plants... if this was so easy and possible, WHY doesnt the bedding plant industry use it? you're going to tell me multi national corporations wouldn't jump on glow in the dark pansies? glow in the dark sanseveria for in-door air purifiers?

page5


  • heres the magic, after those rootballs(which can be divided also with great success)are transplanted into a cytokin/auxin mix, causing wicked like lateral branching and upward bonzai growth in a couple days​




you're introducing a callus culture to a cytokinin/auxin mix? first off, this would never happen in a commercial lab because it wouldnt make sense to do that. there's a reason why stage 2 media contains high levels of cytokinins, to produce shoot growth/multiplication, and auxins are used in stage 3 if you can't make the plant root when hardening off. stage 3 media is skipped because it's EXPENSIVE.

the pdf provided on cannabis biotech...
http://www.scribd.com/doc/14571756/The-Biotechnology-of-Cannabis-Sativa

this doesn't provide the media needed to micropropagate cannabis. they say you can use the same media used for carrots. whatever. any real peer reviewed paper would actually PROVIDE the media recipe. they cant prove it works, and the experiment isn't replicatable. the paper is a waste of time

post 48 on page 5 shows a tub tilted on its side... he talked about using this INSIDE a laminar air flow hood. in a hood, you have air being flown at you at over 90 ft/s. this is to prevent pathogens. with the bottom of the tub facing TOWARDS the filter... it's going to block all the airflow. there's no airflow going over the cultures. if you work over the cultures, pathogens aren't going to get blown towards you and out of the hood, away from your samples. THIS IS POOR STERILE TECHNIQUE
 
A

angrygrower

Member
i dont think im allowed to post contact info. im unemployed if anyone wants to hire me for anything. i have a degree. and stuff. let me know if you have any questions, im pretty far removed learning about TC, but ive been to commercial labs, have hood experience, and will answer questions to the best of my degree.

sorry for multiple posts. this is the top result for marijuana tissue culture. this kind of crap shouldn't stand.
 
O

overmyhead

Well-Known Member
This stuff is overwhelming to me but I'm very thankful I stumbled upon it. Will be coming back! Thank you for sticking with it.

pharmacoping said:
pressure cooker works great, and ul light mutates genetics, usually negatively, and usually irresponsibly. an autoclave is unecessary until you have gallon batches and hundresds of prepared vessels to sterilize.

if easy?? what the hell is easy kiddo? this is not easy. I use it to to put 30 more plants into play every grow cycle, instead of housing clones/moms. lets see...30 x plant value = ahhh, cant be done. you stick with that answer, and move aside as a provider of healthy verifiable pathogen free plant medicine.

many things in life are easy for some and difficult for others. For instance, intelligent technological advancements are "easy" for some to imagine, and very difficult for others to grasp. I would still be living at home if I couldn't grasp knowledge, or wanted an easy way out.
Click to expand...
 
pharmacoping

pharmacoping

Active Member
Closetgardner said:
@pharmacoping,what strain do you prescribe for psoriasis? my cousin suffers from the worst case ive ever seen in real life(not on tele) id love to get something to help her out!! im not in a legal country as you can probly tell!!! and what affect does this strain have on the condition does it help clear it up or stop the itchy pain?? id be very grateful if you can help me with this.CG
Click to expand...
I dont have that disorder, but two past patients did. They used olive oil infused with a higher cbd and topically applied it, as well as dosed daily. its uncomfortable for some folks to eat at first, but the approach was internal and external healing. they claim now to only have these issues in places where they didnt apply, like the scalp.
I would saponate the oil like soap if was me, and use it as shampoo. good luck
 
pharmacoping

pharmacoping

Active Member
[h=1]What Kind of Plants Glow in the Dark?[/h]X
Beth Anderle
A former Army officer, Beth Anderle has been writing professionally for many years and is an experienced freelance reporter. Anderle graduated from the University of Maine with a Bachelor of Arts in international relations and completed a Master of Divinity from Northern Baptist Theological Seminary. Her areas of interest including gardening, genealogy, herbs, literature, travel and spirituality.


By Beth Anderle, eHow Contributor



Glow-in-the-dark plants, more correctly called bioluminescents, are found in different environments throughout the world. Bioluminescence is the emission of light from living things. Does this Spark an idea?
[h=3]Other People Are Reading[/h]





  • [h=2]Jack o'Lantern Mushroom[/h]
    • Jack o'lantern mushroom (Omphalotus olearius) is a yellowish mushroom that resembles some chanterelles. Because of a special enzyme, called luciferase, the gills of the mushroom glow in the dark. This mushroom, like most bioluminescent fungi, is poisonous and will cause cramps and vomiting. This is caused by the presence of the poison illudin in its flesh. A sub-variety, the western jack o'lantern mushroom (Omphalotus olivascens), is found in the southern part of California.

    [h=2]Tsukiyotake[/h]
    • Tsukiyotake (Omphalotus japonicus) is native to Asia and particularly common in Japan. The tsukiyotake is a relative of the jack o'lantern mushroom, and is also poisonous, but can easily be distinguished from a chanterelle


    • Sponsored Links


    [h=2]Ghost Fungus[/h]
    • Ghost Fungus (Omphalotus nidiformis) is a mushroom found in southern Australia, and is generally found growing on dead or dying trees. It is parasitic. They look similar to oyster mushrooms, and can be seen at night as a pale glow on the base of eucalyptus trees in a forest. The cap ranges in color and is often tinted with orange, brownish, grayish, purple or even bluish-black shades; the gills are white. The whole mushroom glows, although the green-glowing gills are the most luminescent. It occurs in the south part of Australia and in Tasmania

    [h=2]Honey Mushroom[/h]
    • Honey mushrooms (Armillaria mellea), and other closely related root rot and wood decay fungi found across North America, Europe and Asia, grow on old and rotting trees, loose wood pieces and on buried roots. The main body of the mushroom does not glow, only the rooting portion of it does. Armillaria is also known as "foxfire."

    [h=2]Panellus Stipticus[/h]
    • Panellus stipticus is a tiny fungus that is found in rotting hardwood all around North America; it is more common in eastern North America than in the West. It has a small tan-to-white cap and white luminescent gills. It is very hardy and, after drying out, is able to revive with enough rain. It can be used as a styptic to stop the loss of blood.




Read more: What Kind of Plants Glow in the Dark? | eHow.com http://www.ehow.com/facts_4924225_what-kind-plants-glow-dark.html#ixzz2LGlefS00

Glow-in-the-Dark Plants are Highlight of International Space Station Science Briefing

CAPE CANAVERAL, Fla. -- NASA will shed light on plant investigations aboard the International Space Station in a briefing at noon EST, Friday, Feb. 5. The briefing from NASA's Kennedy Space Center in Florida will be broadcast live on NASA Television.

The upcoming shuttle mission, planned to launch Feb. 7, will continue assembling the space station so it can be used for continuous scientific research as a national and multinational laboratory.

Microgravity plant growth experiments conducted aboard the station will help prepare for long-duration spaceflights of the future. The use of miniaturized green fluorescent proteins, that glow in the dark, and associated compact imaging systems, may be used to help monitor crop conditions on Earth.

The briefing participants are:
-- Julie Robinson, International Space Station Program scientist, NASA's Johnson Space Center, Houston
-- Robert Ferl, principal investigator of Transgenic Arabidopsis Gene Expression System at the University of Florida, Gainesville
-- Wagner Vendrame, International Space Station National Laboratory investigator for National Lab Pathfinder-Cells at the University of Florida, Homestead
-- Perry Johnson-Green, senior program scientist, Life and Physical Sciences, Canadian Space Agency

NASA has published a new Web feature that provides examples of space station research dividends such as those related to cancer treatment delivery, food poisoning vaccine development, air purification, remote ultrasound tests and more.


[h=2]Gold nanoparticles make plants glow in the dark[/h] // November 8th, 2010 // How Things Work, Recent Research, The Realm of Bizzare
Image by Yellowcloud

Imagine if instead of having sensor lights to illuminate a garden path, you could line it with light-emitting plants. You could stroll along bio-luminescent flower beds, dancing in dappled moonlight and delighting in eerily lit peace, free from the shackles of electricity.
It could be possible with sea urchin shaped gold nanoparticles. Seriously, every time I turn on my computer the world gets more random. Sea urchins, I ask you. In any event, they’re called nano-sea-urchins.
Taiwanese researchers made a solution of gold nano-sea-urchins and dipped into it an aquatic plant, Bacopa caroliniana or blue waterhyssop. The nanoparticles moved into the plant over a day or so, and stayed there for about a month.
When exposed to UV light, the nanoparticles produced blue-violet light which encouraged the chlorophyll inside the plant to make red light. The result? An awesome glowing plant, just add UV.
It’s exciting stuff, there are a lot of excellent uses for light emitting things that work inside plants or animals. If the particles could be attached to a drug we could track exactly where the drug goes over the course of a treatment. You could attach it to proteins and find out where they are located inside a plant. Or you could just have a sweet glow in the dark plant in your house or garden.
Of course, you still need to have that UV source. But what’s wrong with having black light in your house or garden? Just think of the possibilities… You could drink tonic water every day, that stuff glows blue in black light because of the quinine.
Also, if you’ve ever wondered if black light can cause sunburn (as I recently have) here’s the low down. Black light is made of UV light which is close in wavelength to visible light, so it’s quite low energy. This counts as UVA, not UVB which causes most sunburns. Large amounts of UVA (such as those found in tanning beds) can cause skin cancer or premature aging, but the small amount contained in black lights is unlikely to do much damage.
 
pharmacoping

pharmacoping

Active Member
Well I'll be goddamned !! plants are glowing all of a sudden using off the shelf components and a little creativity !!!
 
O

oxanaca

Well-Known Member
hey im glad to see your still around. i just read thru most of the thread.

i have all the equipment needed to do tissue culture, however i need to order some new hormones mine are over 2 years old. heres my setup.

DSCN0122.jpgDSCN0124.jpgDSCN0119.jpg
 
O

oxanaca

Well-Known Member
my tent lab was designed for mushroom cultivation. i will no longer be using the tent as i now have a 12x12 room dedicated to tissue culturing and other mad scientist exploits.

i only have one experiment undermy belt in plant tissue culture and that was with 20-20-20 fertillizer and vitamin pills and shit. about 5 years ago.
i couldnt get the seeds to germinate on the agar but after 1 month in 20 jars i saw no mold or bacteria, so i succeeded in that regard.

i would like to learn how to produce this callus superstart and then see if i cant come up with a procedure to acclimatize them in my air atomized aeroponic misting environment
 
O

oxanaca

Well-Known Member
just ordered

iaa
iba
naa
kinetin
gibberilic acid
all powder form

plus
brassinoid
triacontanol
fulvic acid
humic acid
to experiment with on ex vitro plants

thinking about looking for some TZD as ive heard thats a good one according to a chinese study on hemp
i already have enough MS salts to make 10 liters of media. and enough agar agar to make 300liters.
 
O

oxanaca

Well-Known Member
if you wouldnt mind telling me, what is your disinfestation procedure for cannabis.

i own a ultrasonic cleaner, tween 20, filter flask with hand pump vacuum(in plants from test tubes it mentions that using filter flask with a stopper and applying a 25mm HG vacuum really helps dislodge a lot of bacteria hiding in crevices of the explants.

i also have all the other disinfectants 70% iso, ethanol, hydrogen peroxide, calcium and sodium hypochlorite.
 
pharmacoping

pharmacoping

Active Member
Dude !!! your set up rocks ! I'm watching you closely now. Skip the TZ and the vacuum cleaning, least I do. your jars are clear, and that's a great sign, you've mastered the sterile technique, the most difficult part for most of the people who try. I've seen people work at it for a year , unable to avoid the fungus, and give up !. stick with it man, it is the hydroponics of the highest order . I lie your aeroponic idea. I use sterile water to roughly wash my samples one at a time. Then 70% iso, then water again, then a 5%bleach solution, then water again. Like you, I use an ultrasonic toothbrush cleaner. Since getting one of those I've seen NO molds for a year. I have control vessels more than two years old with no mold present. I recently transferred into a dozen 2 year old prepared test tubes and my developed calli look happy.I no longer use any of the hormones listed in culture. I make willow water and use it sterilized for pretty good success. Although my growth rate is slower I don't mind the couple week delay, trading for a hormone free existence. peace
 
O

oxanaca

Well-Known Member
your not using any hormones? sweet

im actually in the process of moving all my science stuff to my new house where my "lab" is located.

i would rate my aseptic technique as excellent at this point, based on less that 1% of my mushroom cultures contaminating.
it should be ive been doing fungal culture for over 500hours over the last 5 years.

i am in the processs of buying some Isaria fumosorosea. it is a parasitic fungus that colonizes thrips, whiteflys, and most importantly spidermites, and kills them. i will maintain and cultivate this fungus for my own use.

http://www.sepro.com/default.php?page=preferal


also there is a member on here that goes by canndo or something that had some positive micropropagation experiances. im going to start chatting with him if i can to see if he has any advice for me.
 
O

oxanaca

Well-Known Member
if i were to collect the growth tips off my favorite wreckage strain and successfully sterillize them.
what would you recommend i add to my media?

full strength M/S?
grams of sucrose per liter?
what do you do for agar. i use a generic non tissue culture grade agar. for my fungal cultures. and it works good. i know for the M39 MEA agar i usually make for mushrooms i use 20grams per liter. im assuming i want a much lower concentration as the gel strength is quite strong at 20g


im going to try your hormone profiles before i do much experimentation seeing how your method has shown you results.
 
O

oxanaca

Well-Known Member
have you ever used a culture rotator before. this sounds like the bees knees. not sure how this would work for the callus cultures you speak of though

i will not have to use agar for cultures. as a rotator allows for a liquid medium
the roots will never have dissolved oxygen issues, at least due to the presence of a solid agar medium.
and i will save time not having to worry about washing agar off roots.

i will not be able to detect contaminates however, so i would probably just use this for the final multiplication before acclimatization.
 
O

oxanaca

Well-Known Member
pharmacoping said:
after wondering why I haven't met anyone here with mj culture success, a possible answer dawned on me. This is the most combative,immature, misinformed marijuana community I have ever encountered. Some of these same mindsets are, fuckit, ..

I thank the couple that helped me find two forums full of like minded individuals successful in the art(evidently) of marijuana tissue culturing, botany, and horticulture. I am a member in these communities now and will probably never return here, well, unless I'm having a hard time figuring out if my female has been seeded(brown pistils two weeks flower, as suggested here by a member) or if its a male or female.
Click to expand...
hey dude can you point me in the direction of these forums. RIU is not where its at when i comes to plant tc
 
pharmacoping

pharmacoping

Active Member
I dream of rotators man, glad you mentioned them here. I do not own one though. I would suggest a readily available carrot culture recipe for the first time cannabis culture. I purchase commercial tc agar now, but actually used gelatin in the old days. I don't share my cannabis recipe, but after your calli are thriving in carrot culture, unless blind, you'll know exactly what to adjust to meet your expectations, divide, chute, root, or get off the pot right ! :) With a hundred or so practice runs you'll have your own recipe that succeeds, and I guarantee you it's different than mine. cannabis is not as forgiving as houseplants an flytraps. You'll be fast and fine with your my-silly-um experience. remember hormones, less is always better, always, considering your culture can even do without them..i use white cane sugar, by the way. peace
 
You must log in or register to reply here.
Top