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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Micropropagation at home.
#14013111 - 02/23/11 11:53 AM (13 years, 10 days ago) |
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Ok, I was reading here about micropropagation the other day, and wondered why it's so fancy and exotic. Why we don't have home teks. A few PMs later, I'm convinced it can be done.
Well, so today I go down to the local garden shop, I'm looking at the shelves and this employee asks if I need help. "Rooting hormone?" "What are you rooting," she wants to know. "I was reading on the 'net about growing plants in dishes, rather than from seed. Sounded so cool I had to try it." "Oh, you want this." turns around, grabs a bottle off the shelf, hands it to me.
"So this works?" "Oh, yeah, I did it college," she says "it was one of my favorite experiments." "Really? So, I need plant food too right" "Yep. Miracle Grow." Hands me the MG.
"Ok, so... do I want silica gel? Or agar?" "Agar." No hesitation, no uncertainty. "Agar." "Agar."
"So, this really works? For reals?" "Oh yeah, works great"
So I guess I'm going to try it.  Plan for the first run is to mix the MG and root hormone at whatever concentration the labels says is normal, use that to make agar jars. I'll be using PF jars with plastic lids.
Ideally, one would like a plant with a known sex, but I just have some seeds. It's 50/50 odds, right? If I sprout three seeds, I should get at least one female?
-------------------- You make it all possible. Doesn't it feel good?
Edited by Doc_T (02/23/11 12:05 PM)
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El Douche
El Douchius Maximus


Registered: 01/08/11
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Re: Micropropagation at home. [Re: Doc_T]
#14013232 - 02/23/11 12:22 PM (13 years, 10 days ago) |
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I use rooting hormone for house plants. I just cut off a small branch, pluck the bottom leaves, dip the stem in hormone powder - including at least one nub (branching point), and place in tap water. Roots usually grow out in a week or 2. You could also place the dipped cutting in potting soil or other media and put a humidity tent over it untill the plant grows roots (you can use an inverted cut 2-liter as a tent).
I don't know how this will work with your specific plant, as my described method does not work on all of my plants.
You could put some Miracal Grow in the water.
I would think that agar would be difficult because it is prone to contaminants. The hormone used to come with fungicide, but the last time I looked it could not find the combination.
The hormone is toxic, so don't breath the dust or get it on your skin. A very small amount will kill an animal, and it is used in small amounts to kill fish in lakes.
-------------------- I'm The Douche, Doucher, His Royal Doucheness, or El Doucherino if you are not into that whole brevity thing. Trade List : Wanted/have edible cultures, ethobotanicals, cool plants, cacti.
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Djcorbetto
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Registered: 01/12/11
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Re: Micro propagation at home. [Re: El Douche]
#14013387 - 02/23/11 12:54 PM (13 years, 10 days ago) |
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sterilisation is very important once you've got that sorted then you should be ok. Ive been waiting for this, will see if it works compared to the normal route pretty much everyone Ive read up on has used which is getting all the chemicals and making their own media up and using M&S media
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Micro propagation at home. [Re: Djcorbetto]
#14013413 - 02/23/11 12:59 PM (13 years, 10 days ago) |
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Sterile technique I have down, it's fundamental to mycology. 
Completely unrelated question, there was a cactus name Trichocereus transchekii, or similar. Is that one I want to back and buy?
-------------------- You make it all possible. Doesn't it feel good?
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Djcorbetto
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Re: Micro propagation at home. [Re: Doc_T]
#14013460 - 02/23/11 01:08 PM (13 years, 10 days ago) |
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Done a bit of googling at Trichocereus terscheckii seems to be the one. on erowid it says that anyway.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Micro propagation at home. [Re: Djcorbetto]
#14013478 - 02/23/11 01:11 PM (13 years, 10 days ago) |
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That's a good one?!?!
Edit- apparently nonmagical but a fun cactus anyway. I may get it if it's there next week, but not worth driving back for.
-------------------- You make it all possible. Doesn't it feel good?
Edited by Doc_T (02/23/11 01:20 PM)
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caricapapaya
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Re: Micro propagation at home. [Re: Doc_T]
#14014039 - 02/23/11 02:33 PM (13 years, 10 days ago) |
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If it doesnt work, try using the hormones at way lower conc than suggested on the label.
You'll need to look at the active ingredient (probably IAA, NAA, IBA or a combination) and then do some calcs to bring the level to µM levels...
try it out. you have sterile technique down, you shouldnt have a problem.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Label says IBA.  I used just a knifetip of it, my scale won't measure this small. Anybody know whether the stuff is even PCable?
-------------------- You make it all possible. Doesn't it feel good?
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caricapapaya
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Re: Micro propagation at home. [Re: Doc_T]
#14014333 - 02/23/11 03:08 PM (13 years, 10 days ago) |
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yes, it can be autoclaved.
also, if you can dissolve it something, you can make a concentrated stock by weighing out more than you need, and then dilute that by volume instead of weight
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brshroomer
Moss bear hunter



Registered: 12/10/06
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Re: Micro propagation at home. [Re: Doc_T]
#14014348 - 02/23/11 03:10 PM (13 years, 10 days ago) |
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research into forming callous cultures, these can be split over and over to make hundreds of cultures, they grow under a specific hormone(i can't remember which now)
then you chance the most proeminent hormone on the cultures and they will form plantlets, which can be extracted from the agar just as normal seedlings.
i had some papers on it, i'll see if i can find them.
also, look into murashige skoog medium, it's a very basic 'good for all' medium.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Micro propagation at home. [Re: brshroomer]
#14014371 - 02/23/11 03:14 PM (13 years, 10 days ago) |
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Quote:
brshroomer said: murashige skoog medium, it's a very basic 'good for all' medium.
I'm hoping this will be a local-ingredients grow. I don't want to send out for anything. Several reasons, biggest being money. I'm just goofing around here, I don't plan to really grow anything much. So sending out for supplies makes no sense.
But also I just think it's cooler if you can do it with regular OTC stuff.
-------------------- You make it all possible. Doesn't it feel good?
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brshroomer
Moss bear hunter



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Re: Micro propagation at home. [Re: Doc_T]
#14014430 - 02/23/11 03:21 PM (13 years, 10 days ago) |
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i agree that OTC ready mixed nutrients are WAY cheaper and practical, but i have to test them yet, i don't know if the proportions in the solution are right.
if it does work that may be a breakthrough for home tissue culture.
also, consider adding dextrose to the medium, it is added in MS medium, and it's gotta be there for some reason.
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Micro propagation at home. [Re: brshroomer]
#14014455 - 02/23/11 03:24 PM (13 years, 10 days ago) |
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Oh, I wondered about dextrose. Next time maybe. I'm sprouting two seeds, that gives me 75% chance of at least one female.
-------------------- You make it all possible. Doesn't it feel good?
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Burbles
Stranger


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Re: Micro propagation at home. [Re: Doc_T]
#14014543 - 02/23/11 03:36 PM (13 years, 10 days ago) |
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Sucrose is better from what I have read...
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Micro propagation at home. [Re: Burbles]
#14014548 - 02/23/11 03:37 PM (13 years, 10 days ago) |
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Table sugar? Really? Weird.
-------------------- You make it all possible. Doesn't it feel good?
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Burbles
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Re: Micro propagation at home. [Re: Doc_T]
#14014665 - 02/23/11 03:57 PM (13 years, 10 days ago) |
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Yes - this actually makes sense, that is the main sugar used within plants, and that also explains why sucrose is so available, we get it from plants. Other than that, I don't know why, but it is what I have read.
I was thinking today in class that propel - that gaterade stuff, might actually be a good thing to add to these experiments, as it has EDTA - which is a common chelating agent and would help the metals stay in solution better, in addition it also has small amounts of various B vitamins (some are needed by plants, IIRC - like thiamine?) and has other things like citric acid (its used in MicroP for various reasons, as a nutrient, pH regulation, etc) and ascorbic acid (another chelating agent, if I remember correctly - though probably would decompose on PCing)
Might be worth a shot, I don't know.. I think EDTA is important though, and is hard to find OTC unless it is in one of those gunky EDTA supplements...
Also: Link & description about Banana micropropagation and other exotic fruit/plants http://www.thelaboratory.org/talk/index.php?topic=134.0 Amamature Micropropagation thread in general.. has some useful info... http://www.thelaboratory.org/talk/index.php?topic=18.0 First attempt at micropropagation... using potato cuttings, guano tea coconut and sea water http://www.thelaboratory.org/talk/index.php?topic=202.0 And also there is a bit more about growing out cacti callus cultures, etc.. but i don't want to spam my site - but there is a pretty good amount of micro/plant tissue culture info there Also check out these micropropagation videos, great for beginners using the Home plant tissue culture materials.. http://www.youtube.com/user/fbt2007
glad you are trying some stuff out!
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cc2
Mush

Registered: 05/15/10
Posts: 2,611
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Re: Micro propagation at home. [Re: Burbles]
#14014710 - 02/23/11 04:06 PM (13 years, 10 days ago) |
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good luck! 
IBA stands for indole butyric acid, not bad rooting hormone at all if you've got powder form it's better 'cos solutions don't store for very long even if refrigerated
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Burbles
Stranger


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Re: Micro propagation at home. [Re: cc2]
#14014768 - 02/23/11 04:16 PM (13 years, 10 days ago) |
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Quote:
GIBBERELLIC ACID (GA3)
Stock solution of gibberellic acid: I ,000 ppm 1. Weigh 0. 2 p of gibberellic acid and dissolve well with some alcohol drops. Add 200 ml of distilled water 2. Keep in a conveniently labeled vial at 0°C. The gibberellic acid may be sterilized together with the culture medium: however, the loss of some activity is also possible. One ml of concentrate solution (1,000 ppm) contains 1 my of gibberellic acid.
NAPHTHALENE ACID (NAA)
1 Stock solution of NAA 1,000 ppm Weigh 0.2 g of NAA and dissolve well with some NaOH 1N drops. 2. Add 200 ml of distilled water. Keep it in a conveniently labeled vial at 0~O. One ml of stock solution (1,000 ppm) contains 1 mg of NAA.
N-6-BENZYLAMINOPURINE(BAP)
Stock solution of BAP: 1,000 ppm 1. Weigh 0. 2 g BAP and dissolve well with some drops of NaOH 1N. Add 200 ml distilled water. 2. Keep in a conveniently labeled vial at 0°C. BAP may be sterilized together with the culture medium; however, the loss of some activity is also possible. One ml of stock solution (1,000 ppm) contains 1 mg of BAP
INDOLEACETIC ACID (IAA)
Stock solution of IAA: 1,000 ppm 1. Weigh 0.2 mg of IAA and dissolve well with some alcohol drops. Add 200 ml of distilled water. 2. Keep it in a conveniently labeled vial at 0°C. Sterilization by filtration is recommended. One ml stock solution (1,000 ppm) contains 1 mg of IAA.
KINETINE (KIN)
Stock solution of KIN: 1,000 ppm 1. Weigh 0.2 g KIN and dissolve well with some drops of NaOH 1N. Add 200 ml of distilled water. 2. Keep in a conveniently labeled vial at 0°C. KIN may be sterilized together with the culture medium; however, the loss of its activity is also possible. One ml of the stock solution (1,000 ppm) contains 1 mg of KIN.
2,4-DICHLOROPHENOXYACETIC ACID (2,4-D
Stock solution of 2,4-D: 1,000 ppm 1. Weigh 0.2 g of 2,4-D and dissolve well with some alcohol drops. Add 200 ml of distilled water. 2. Keep in a vial conveniently labeled at 0°C. 2,4-0 may be sterilized together with the culture medium; however, a loss of its activity is also possible. One ml of the stock solution (1 000 ppm) contains 1 mg of 2,4-0.
The actual water solubility of many of these compounds is very low - and sometimes will not fully dissolve in water if you don't use the above methods.
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Djcorbetto
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Re: Micro propagation at home. [Re: Burbles]
#14015155 - 02/23/11 05:29 PM (13 years, 9 days ago) |
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I had a recipie for shoots for cannabis, and it just said: M&S media with 15g/litre of sucrose. when i spoke to carol at home tissue culture she reccomended that i got the M&S media with vitamins as its same price but better, ive read that if you add a multivitamin tablet to the solution that helps too
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Burbles
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Re: Micro propagation at home. [Re: Djcorbetto]
#14015699 - 02/23/11 06:57 PM (13 years, 9 days ago) |
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It didn't have any hormones at all?? And it was for shoot formation, and not root formation?
The root formation would make a ton more sense to me, they root naturally - but it wouldn't make sense to have it grow more shoots and no roots if it is just a small random piece of flesh...
But I guess if it were a stem, it could easily grow both, esp. if it had a tip that would allow for leaves to grow out already..
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