Agrobacterium rhizogenes -- How to obtain?

[Vesp]

Agrobacterium rhizogenes is the bacteria that causes hairy root disease, this has some uses, as has been discussed else where on this forum. I am interested in a method in obtaining Agrobacterium rhizogenes.

The only source I've found is this one, and it costs $155.00 dollars, which for bacteria, is just a little bit over priced, I think.

http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=11325&Template=bacteria

How about some method of getting it from the soil? I know most Agrobacterium bacteria will swim towards phenolic compounds, since this generally implies roots of plants have been injured, and that allows them to enter the plant and cause problems such as hairy root disease or crown gull.

I think getting them this way is a slim chance to none -- I assume making a tea of some fairly phenolic root, or just a tea of some plant in general ought to bring them out if they exist, but then I would probably need to isolate them. I doubt they are even present in everyday soil.

 Does anyone have information on how it can be obtained from the wild, or perhaps know a better source for this bacteria? I am thinking that I might just have better luck contacting a professor or a college student and get it this way -- but I don't know if they would be willing to do that or not.

Maybe if life is great, you can just add the phenolic compounds, get the soil which presumably has them in it, and then infect a plant with them by harming its roots with the 'tainted' soil present. Anyways.. some articles on how they are obtained by nature or a cheaper source would be great to see! I can't find much on them at all.

PS. This may prove to be rather difficult since  the places that sell Agrobacterium tumefaciens claims it has a restriction on it, and that you need a USDA permit. I.e: http://www.carolina.com/product/living+organisms/prokaryotes/bacteria+cultures+and+sets/acetobacter+aceti%2C+living%2C+tube.do?keyword=Agrobacterium&sortby=bestMatches&page=1

However, they are only selling it for $9.50

Plus: http://www.carolina.com/product/plant+cancer+study+kit.do?keyword=Agrobacterium&sortby=bestMatches

Edit:
While this isn't the Agrobacterium rhizogenes, which is wanted, it contains some information about the isolation of A. Tumefaciens, which might prove to be useful later on.

A. tumefaciens can be effectively isolated for identification from gall tissue, soil or water. Optimal gall tissue for isolation is white or cream-colored from a young, actively growing gall. The gall should be washed or surface sterilized using 20% household bleach, and rinsed several times in sterile water. Cut a few samples from different parts of the white tissue of the gall, and further divide samples into small pieces. Place these pieces into a culture tube containing sterile distilled water or buffer, vortex and allow to stand for at least 30 minutes. Using an inoculating loop, streak this suspension on Medium 1A (Schaad et al., 2001), and incubate at 25-27° C. Different strains will grow at different rates. One may also use this selective medium to detect A. tumefaciens in soil dilutions or irrigation water.

It should be noted, however, that the presence of A. tumefaciens cells in a sample does not necessarily dictate the existence of the crown gall-inciting strain in the sample. Only cells containing a specific plasmid (the Ti plasmid) can cause disease. A. tumefaciens strains lacking the plasmid live as rhizosphere-inhabiting bacteria without causing disease.

--- http://www.cals.ncsu.edu/course/pp728/Agrobacterium/Alyssa_Collins_profile.htm

[Naf1]

Your last comment was the best I think, there would be several ways to go about it one would be to find a species of plant that is susceptible to the disease. Plant it in the soil and try and induce the disease by damaging roots, keeping the soil soaking wet 24/7 , putting a bag over it so the humidity raises up to near 100%, ect. If you can induce this disease in a plant you have a nearly limitless supply of the bacteria, you can make substrates that can be used to culture the bacteria; Agrobacterium Agar cultures. Then cutting open a tumor and taking a sterile tissue sample to grow on your substrate. Even following the above method as stated you can just use water for microscopy purposes. But to actually grow it you need the agar cultures above.

Protocol for isolation of Agrobacterium from herbaceous plant material
http://www.science.oregonstate.edu/bpp/Plant_Clinic/Protocol%20for%20isolation%20of%20Agrobacterium.pdf

Isolation of Agrobacterium tumefaciens Biovar 3 from Grapevine Galls and Sap, and from Vineyard Soil
http://www.apsnet.org/phyto/PDFS/1983/Phyto73n02_163.pdf

Shelf-stable, virulent preparation containing Agrobacterium cells, an acidulant and a phenolic compound
http://www.freepatentsonline.com/6540997.html

Infecting growing tissue with agrobacterium;

     
                                                                                                                       Plant (S. chacoense) transformed using Agrobacterium. Transformed cells start forming calluses on the side of the leaf pieces

"This may prove to be rather difficult since  the places that sell Agrobacterium tumefaciens claims it has a restriction on it, and that you need a USDA permit"

As you stated this is pretty full on business, genetic modification is a little taboo at the moment. Think about it, companies like Monsanto are not allowed to produce GM crops for food as no one knows if it will be bad for us down the road (they have started doing it anyway, but if they told everyone which products where GM at the moment they would stop buying them so is kept secret). But the second the theory was introduced (way before people started thinking about the moral and maybe physical ramifications that come from it)companies like Tasmania Alkaloids started using it to produce ALL of their product so Codeine, Morphine, Thebaine and the products derived from them (which is too many to list, but includes Oxycontin, Oxycodeine and all the popular opiates) which are put directly inside the body IV or Patches or tabs, the opiate medication on offer today is 100% genetically modified(maybe with the exception of that produced in Turkey, but it would be suprising to hear they were not keeping up with the Jones's), coffee another example, tobacco another. If people think they are not consuming GM products because they are not buying canola oil (they are kidding themselves). Also if your immune system is not up to scratch and you are in contact with a lot of it, it can actually start infecting humans. And you can imagine ;

"There is a conjectured connection with Morgellons syndrome. Dr. Stricker, along with Dr. Citovsky, MRF board member from the State University of New York at Stony Brook and an expert on plant pathogens, reported in January, 2007, that Morgellons skin fibers appear to contain cellulose. Five skin samples of Morgellons patients contained evidence of DNA from Agrobacterium.[4]"

And thats without the tumors! So care should be taken not to infect yourself or anything you are going to consume! Without proper analysis you should not consume any foodstuffs prepared with those methods, as you may have inadvertently made it more carcinogenic for example. A basic example would be the most simple food; Lettuce like all plants, contains a small amounts of caffiec acid (which is a known carcinogen) if you were to push the levels of caffiec acid through the roof with agrobacteria but did not know this was a consequence the subsequent lettuce produce would be significantly more carcinogenic than the normal stuff! There are examples like that which can be made for nearly any vegetable/crop, although probably not likely it is an example of why it should be thoroughly tested.

The easiest way is going to be to buy some though, you will get the right strain for sure, it will be sterile and ready to go.  You could infect on a petri dish(plate culture);

Isolation of novel strains of Agrobacterium radiobacter with altered capacities for lactose metabolism and succinoglucan production
Journal of General Microbiology (1990), 136, 2179-2188.
http://mic.sgmjournals.org/cgi/reprint/136/11/2179.pdf

Agrobacterium rhizogenes-mediated transformation of opium poppy, Papaver somniferum L., and California poppy, Eschscholzia californica Cham., root cultures
http://jxb.oxfordjournals.org/cgi/reprint/51/347/1005.pdf

Or using the floral dip method;

Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana
http://faculty.salisbury.edu/~flerickson/protocols/floral%20dip%20transformation.pdf

[Vesp]

Thank you, great reply and very interesting! I looked into that Oregon state university protocol, as well as the A1 and 2E culture mediums -- that doesn't seem to hard to do at all.

However, with the  Agrobacterium rhizogenes doesn't produce a tumor, like the Agrobacterium tumefaciens, it just produces a lot of roots -- it would be very hard to tell if the plant in the ground has an abnormal amount of roots or not... It seems like you'd need to hard the plant -- Inject the suspected material into the plant to see if it begins having hairy root disease. This would make it much more time consuming and problematic.

However, the more I think about it, the less I think you'd need a sterile supply of the A. rhizogenes if the goal is to simply get a few plants infected with the hairy root disease... simply because most other bacteria isn't really going to affect it, you'd most likely just have to watch out for A. tumefaciens being present at the same time, as that may cause some problems with the wanted disease. I bet they are probably pretty hard to separate from one another.

What you've posted here, is one of the most amazing things ever if I am understanding it correctly.

"There is a conjectured connection with Morgellons syndrome. Dr. Stricker, along with Dr. Citovsky, MRF board member from the State University of New York at Stony Brook and an expert on plant pathogens, reported in January, 2007, that Morgellons skin fibers appear to contain cellulose. Five skin samples of Morgellons patients contained evidence of DNA from Agrobacterium.[4]"

[Naf1]

"What you've posted here, is one of the most amazing things ever if I am understanding it correctly."

That they can genetically modify humans, with weakened immune systems also. The whole quote from wiki'

"Although generally seen as an infection in plants, Agrobacterium can be responsible for opportunistic infections in humans with weakened immune systems,[1][2] but has not been shown to be a primary pathogen in otherwise healthy individuals. A 2000 study published by the National Academy of Sciences suggested that Agrobacterium attaches to and genetically transforms several types of human cells by integrating its T-DNA into the human cell genome. The study was conducted under laboratory conditions and states that it does not draw any conclusions regarding related biological activity in nature.[3]
There is a conjectured connection with Morgellons syndrome. Dr. Stricker, along with Dr. Citovsky, MRF board member from the State University of New York at Stony Brook and an expert on plant pathogens, reported in January, 2007, that Morgellons skin fibers appear to contain cellulose. Five skin samples of Morgellons patients contained evidence of DNA from Agrobacterium.[4]"

Contribution of Agrobacterium to Morgellons Disease.

Skin biopsy samples from two Morgellons patients were subjected to high-stringency PCR testing for genes encoded by the Agrobacterium chromosome. Screening of the same samples for Agrobacterium virulence (vir) genes and T-DNA sequences in the patient's genome was also performed. Results: PCR screening indicated the presence of Agrobacterium genes derived both from the chromosome and from the Ti plasmid, including the T-DNA, in tissues from both Morgellons patients. Conclusions: Our preliminary results indicate that Agrobacterium may be involved in the etiology and/or progression of Morgellons disease. If these results are confirmed, it would be the first example of a plant-infecting bacterium playing a role in human disease.

http://www.morgellons.org/suny.htm

Agrobacterium infections in humans: experience at one hospital and review.

http://www.ncbi.nlm.nih.gov/pubmed/8448285

Agrobacterium & Morgellons Disease, A GM Connection?

Preliminary findings suggest a link between Morgellons Disease and
Agrobacterium, a soil bacterium extensively manipulated and used in making GM
crops; has genetic engineering created a new epidemic?

http://www.morgellons-research.org/morgellons/agrobacteriumAndMorgellonsFull.pdf

[Naf1]

"However, with the  Agrobacterium rhizogenes doesn't produce a tumor, like the Agrobacterium tumefaciens, it just produces a lot of roots -- it would be very hard to tell if the plant in the ground has an abnormal amount of roots or not... It seems like you'd need to hard the plant -- Inject the suspected material into the plant to see if it begins having hairy root disease. This would make it much more time consuming and problematic."

Ohh yeah, I was interested in A. tumefaciens so was talking about that (oops), You could always grow it in a pot, make a clear one, or simply hold the stem and push the pot off and inspect the roots manually. Then put it back in the pot, in one of those papers above they use the tip of a toothpick to absorb the colonized water then dry it and store them wrapped in sterile aluminium foil until ready to use. When ready they unwrapped the foil and pushed the tip of the toothpick into the plants stem and left it for a week and then took it out.

Clear Orchid Plant Pots

http://www.repotme.com/orchid-pots/Orchid-Pots-Clear.html

Just put that inside another black pot, so the roots dont get to much sunlight! When you need to check, just take the clear pot out of the black one and look.

[Vesp]

Good idea, but I was also kind of worried that it may not be that easy to identify normal root growth vs hairy root growth. I'll just try to keep a control if I ever really get around to doing this.

The A. tumefaciens is a great bacteria as well, and would have more use in genetically modifying a plant by horizontal gene transfer, but this I feel is much to far out of reach vs just giving a plant a disease and growing the roots in a neutrient solution. It would probably be a close to impossible task to take a gene from one plant, put it in the bacteria, and then successfully get that to take into another plant - then isolate.
Hopefully I am wrong about that, but isolating a wanted gene is not easy as far as I know.

[Naf1]

All Agrobacterium use horizontal gene transfer, their plasmids (either tumor inducing plasmids or rhizome inducing plasmids respectively) all work the same way on a molecular level (they can use Ti or Ri plasmids to insert small pieces of their T-DNA into the plants chromosomal DNA, which is incorporated at a semi-random location in the plant genome.) Thereby changing the plants DNA, chromosomal DNA is responsible for many functions including regulatory ones [Boveri proved that chromosomes were the vectors of heredity!] If that diseased plant is then used to make seeds the copy of its DNA it passes on have been changed by recombinant DNA by the bacteria thereby producing progeny with modified genes that the bacteria changed (a generation before). Changing the chromosomal DNA of the plant can have drastic effects in the progeny, such as modulated alkaloid biosynthesis.

Agrobacterium rhizogenes, also uses the same techniques but has specific Ri plasmids (Rhizome inducing) opposed to the Ti plasmids (tumor inducing) in A. Tumefaciens.

"Hopefully I am wrong about that, but isolating a wanted gene is not easy as far as I know."

No you are dead on about that, thats why the above can do the job more randomly and much easier. Then when for example you do it, you would infect many plants and make seeds of them and test for increased potency, if they are producing other alkaloids ect. Then after a long time if you strike gold, and say find a thebaine producing poppy amongst your test field. Then you Take a sample and test its DNA, thats those pages of random looking digits in the patents. Then when you have that data you can embark on producing those poppies using recombinant DNA techniques like cloning and chimeric plasmids.

Genetically modified bacteria, where for example," Herbert Boyer working at a University of California laboratory took a version of the human insulin gene and inserted into the bacterium Escherichia coli to produce synthetic "human" insulin."

Is another barrel of monkeys all together!

[Vesp]

I took a walk down to the park to see if any trees seem to have any tumors -- hopefully being caused from the A.T. Bacteria -- I found a bunch of them, so I decided I'd mess around with it a bit and take a few living pieces from  a large chunk of wood coming out of the ground.

After I cut some of the pieces out I decided I'd jab the blade into another tree or two and see if anything starts to grow into a tumor. It would be kind of neat if it did.


Edit: Is my understanding correct that the only reason the agrobacterium tumifaceins does this is to have the plant release opines, which it uses as a source of nitrogen, and fuel?

[Naf1]

From your post;

A. tumefaciens can be effectively isolated for identification from gall tissue, soil or water. Optimal gall tissue for isolation is white or cream-colored from a young, actively growing gall. The gall should be washed or surface sterilized using 20% household bleach, and rinsed several times in sterile water. Cut a few samples from different parts of the white tissue of the gall, and further divide samples into small pieces. Place these pieces into a culture tube containing sterile distilled water or buffer, vortex and allow to stand for at least 30 minutes. Using an inoculating loop, streak this suspension on Medium 1A (Schaad et al., 2001), and incubate at 25-27° C. Different strains will grow at different rates. One may also use this selective medium to detect A. tumefaciens in soil dilutions or irrigation water.

http://www.cals.ncsu.edu/course/pp728/Agrobacterium/Alyssa_Collins_profile.htm

Grow a culture Vesp! You need medium 1A, do you have a recipe?

[Vesp]

I was planning too grow a culture, but I thought it would be interesting to see if I could infect a tree that easily.   I mostly got the top white live area.. and if not, I can always get more... After all, I also got this guy, which will probably be a better source for the bug anyways... (see bottom picture)


Yeah I have a recipe for medium 1A -- http://www.science.oregonstate.edu/bpp/Plant_Clinic/Medium%201A.pdf as has been linked before in this thread -- but I do not have all of those chemicals.

Which are:

arabitol  -- hopefully this can be substituted, as I don't have this or know where to get it. I figure a normal sugar would work, but if not at least another sugar alcohol could -- any suggestions?
NH4NO3 -- I have this,
KH2PO4
K2HPO4 ... as for the phosphates, I currently only have Na3PO4 and some nasty soapy phosphoric acid.
Sodium taurocholate -- I have sodium cholate or perhaps it was deoxycholate? Either way, this is just to act as an emulsifier AFAIK, so I think it is substituted for that.

MgSO4 ·7 H2O  -- of course I have this...
Agar ... also have this...
Crystal violet, 0.1% (w/v) aqueous ... wtf is this?!
*0.1% = 0.025 g crystal violet in 25 ml deionized water.

2 % cycloheximide** 250 ?l 500 ?l 1.0 ml  
1% Na2SeO3 (anhydrous)+ 1.65 ml 3.3 ml 6.6 ml
I don't have either of the last two... perhaps they are not important, considering how little of it there is?

Any suggestions, or better more OTC recipes?

Looks like it is time to dig out some of my culturing setup... if I can find it all :S


PS. I should be fine with freezing the bacteria/plant matter till I get everything going, right?
 

EDIT:

Perhaps I don't need to worry about obtaining that hairy root disease after all. The only reason I had an interest in it is to grow roots that would provide me with useful secondary metabolites.
However, if I am reading this correctly, it seems like you can just culture the grown galls and obtain secondary metabolites as well. I don't know if this is as effective but I don't see why it wouldn't be?

However, this might also cause a lot of opines to be produced, they might turn out to be a problem, but I doubt it. The other down fall is that unlike the hairy root disease, functional plants cannot be grown from the culture, while a plant that is fully capable of living by itself can be made from the roots of the agrobacterium rhizogenes

Crown gall culture and production of tanshinone in Salvia miltiorrhiza.

Zhang Y, Song J, Zhao B, Liu H.

Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Beijing, China.
Abstract

Crown galls were induced by direct infection of sterile seedlings with Agrobacterium tumefaciens C58, and the transformation was proved by opine identification. The crown galls grew well in a hormone-free medium. B5 and MS basic media were good for the growth of crown galls (strain Ca), which increased up to 102 and 90 times in a month, respectively. However, 67-V and WP basic media are good for tanshinone production. It has been shown that crown galls can be utilized as a culture system to produce secondary metabolites.

--- http://www.ncbi.nlm.nih.gov/pubmed/8562850

[Vesp]

All Agrobacterium strains should be able to grow on complex media. A survey of the literature indicates that Agrobacterium is frequently grown in Luria-Bertani (LB), nutrient broth (8 g/l), with or without the addition of yeast extract or glucose, and many other complex media. Potato dextrose agar plates are a common medium for the growth of Agrobacterium.

LB medium (for Agrobacterium growth): 10 g bacto-tryptone, 5 g bacto-yeast extract, 5 g NaCl, pH to 7.5 with 15 g agar for solid media.

-- http://adl.serveftp.org/papers/pyroelectric-ion-acceleration/Superkuh.html

Might be easier to mix up a simple more OTC media.  I am thinking so...

Potato dextrose agar (FDA M127)(abbreviated "PDA") and potato dextrose broth (abbreviated "PDB") are common microbiological media made from potato infusion, and dextrose  (corn sugar). Potato dextrose agar is the most widely used medium for growing fungi and bacteria which attack living plants or decay dead plant matter.[1]

Potato infusion can be made by boiling 300g of sliced (washed but unpeeled) potatoes in water for 30 minutes and then decanting or straining the broth through cheesecloth. Distilled water is added such that the total volume of the suspension is one litre. 20g dextrose and 20g agar agar powder is then added and the medium is sterilized by autoclaving at 15psi for 15 minutes.[2]

A potato, corn syrup, and agar agar sound like the easiest culture -- I think I will use it, unless their is some reason to really focus on the 1A medium?

[Naf1]

re;The agar recipe above you have most of it, arabitol is a sugar alcohol and will be metabolized by the bacteria. You should be able to swap with other sugar alcohols but all are available OTC for use in foods, xylitol, mannitol, sorbitol  and are pretty common and should be able to be found at the supermarket and can be replacements. Dont forget to sterilize it!

Products like this

http://www.shopsafe.com.au/merchants/xylitol_com_au/xylitol.htm

Xylitol, Australian Xylitol Distributor, Large Range Of Sweetlife 100% Xylitol Products
sweetlife® 100% NATURAL XYLITOL products.
sweetlife, a wholly Australian owned company, offers a selection of Natural, Sugar Free, Gluten Free, Low GI products using Xylitol, a natural sugar free sweetener extracted from plants such as corn cobs and birch trees. The body also produces Xylitol during normal metabolism. Xylitol has a low glycemic index of 7, compared with sugar which has a glycemic index of 65. All Sweetlife products have unique packaging that ensures locked in freshness.

You can see why the bacteria would metabolize those specific sugar alcohols with the comment highlighted above. I am sure you could find similar arabitol products as its one of the sweetest sugar alcohols, they usually sell tabs like nutra sweet plus actual granular product to put on you cereal ect.
 

[Vesp]

Good point, but I would still need to find those other ingredients and some seem like it  just isn't worth the battle. Honestly, I don't think I even need to purify and culture the A.T.  -- just need to rub it onto some plants and watch a tumor grow. Once this happens, take the tumor, kill the bacteria, cut it up, and then culture it in a plant tissue culture -- which I can't imagine being anything more then hydroponic fertilizer + sugar.

Hopefully a few of my test plants will prove me correct about this -- then I'll just have to figure out if sassafras is decent to grow as crown gulls. Not that I need sassafrass crown galls for anything. It would just be interesting to do for the sake of doing it.
Plus, it might not prove to be nearly as impractical as I think it is going to be...

[Vesp]

http://www.genscript.com/custom_plasmid_preparation.html

One day this might be a useful link. I'll post it just in case.

Custom plasmids.. So, I assume you can have whatever gene you want inserted into them, and have them shipped to you. These then can be taken up by bacteria, and then inserted into something else -- such as a plant.

[Sedit]

Vesp Im posting this here real quick because I want you to look at it I think youll love. Later is can be deleted bt for download sake here it is....

BTW its pretty large and if theres any issuesextracting it PM me and ill tell you how to getaround the sop called invalid file names

[Vesp]

Thanks! I actually used to have this, but I either misplaced it, or deleted it. I sometimes talk to the guy who compiled it, or at least helped compile it. I talked to him yesterday even. Kanzure is very much into DIY Bio stuff, and he is also the guy who runs http://heybryan.org/



It would be nice to find someone at a university that had access to Agrobacterium rhizogenes -- it is still by far the better bug. IMO

[Vesp]

All right.. so this is a bit off topic, but I am pretty sure someone would enjoy seeing the simplicity in being able to make gentically modify bacteria to glow in the dark, here is a kit for it:

http://www.carolina.com/product/life+science/biotechnology+kits+&+materials/transformation+and+advanced+techniques/glow+in+the+dark+transformation+kit.do

It is likley if the right genes are had from other plant, etc materials that the E coli would also easily take those up as well...

Just need to find a way to isolate wanted genes, and get them to take it up and become fairly functional.

[Locked]

It would be nice to find someone at a university that had access to Agrobacterium rhizogenes -- it is still by far the better bug. IMO

If this is a known "hairy root" tree disease, why not make up a story about your dissertation and contact every arbolist/tree surgeon/tree removal/tree transplanter you can find in the phonebook? Seems like a $20 bird dog payment for each good sample would get you more then you ever wanted, probably delivered to your door. If it is out there, the professionals dealing with trees everyday see it and know it.

I would make the story about protecting the trees from it. I could imagine the normal person being troubled by DIY genetic modification experiments.

Does this just make the roots go crazy, or can one make other bugs hitch a ride in the dna with this tech?

Do you think you could get botulism inside things with this? Botulina toxin is awesome.

Or how about pulling a little magic mushroom hairy root infection on psychedelic cactus? or anything edible for that matter.

[Sedit]

Does this just make the roots go crazy, or can one make other bugs hitch a ride in the dna with this tech?

The bacteria contains DNA strands called plasmids which are naturaly seperate from the nucleus DNA and circular in shape. There are two main types of plasmid from what I gather, one produces Tumors and the other produces Root induction. The root inducing plasmid is what is wanted here. When the bacteria infects the normal root the Plasmid DNA has the ability to penetrate the cell wall of the normal root cell. If its the tumor inducing Plasmid you just get an over growth of plant material that lacks structure. If the Root inducing plasmid gets into the cell and randomly splices itself into the DNA what you get is in essence a tumor of root growth.

If one where to gain the ability to synthesis plasmids from desired isolated DNA strands and inject them into bacteria like the DYI glowing bacteria kit Vesp posted does then the effects produced could exceed many people heres imaginations. The addition of the plasmid to the bacteria does not seem hard at all but specific Plasmid synthsis is where the hard Genetic modification starts and where science is at the for front of just starting to trickle down to amature scientist likely to be placed under restriction sooner or later due to fear and stupidity.

Or how about pulling a little magic mushroom hairy root infection on psychedelic cactus? or anything edible for that matter

Not every plant is infected correctly with Agrobacterium rhizogenes which is depressing although Im possitive there are other means out there that would be what your looking for. Instead of hairy root for mushrooms or cacti I would think you would want the TI plasmid to produce wild growth of the cactus instead of controlled root growth.


I worked in an abandon building recently and the floor has since been covered in about 4-8 inches of "soil". Its dark, damp, and the perfect enviroment for bacteria and fungi. Matter fact I think I have caught a nice chest infection from working there. However on digging thru this I would come to roots whose growth was pretty straight forward then out of no where you would see a hitch in the growth and explosion of microroots from the area. This is a sign of the Agrobacterium rhizogenes bacteria infection and if I where to begin attempts to culture it I know exactly where to look I believe. Its as though the tiny micro roots you normaly see on the side of a root is all that grows and it just becomes a large mass of root growth.

I could imagine the normal person being troubled by DIY genetic modification experiments.

I agree however the trouble would arise if someone where going for something specific. If you want "controlled" random mutations like that of the hairy root infection then most of the hard work has been accomplished by mother nature and its a matter of finding the bacteria to do the job your looking for without any "Real"genetic modification done by youat all. The only skill that should be mastered is tissue culturing since that could pose issues to some. Since rouge DNA can "wiggle" its way into cells im very curious to attempt to lyse some firefly mush and see if any bacteria that grows on it incorperates this DNA into there cells. I doubt It will work but it would be exciting no the lest to see this pile of lysed mushed up firefly cells begin to glow from bacteria digesting and sometime randomly incorperating the DNA of the firefly.

[chaotic_mind]

I work with agrobacterium everyday. AGL1. it stinks! use it to transform plants. was thinking of transforming psilocybe mushroom with a highly active tryptophan decarboxylase gene from madagascar periwinkle but haven't gotten around to it. hopefully will one day.