Tillandsia usneoides tissue culture and transforamtion

[Nico B.]

Has anyone done any tissue cultures or transformations with Spanish moss? looking for a start on how to crow in culture and eventually transform to express some bioluminescent protein that has yet to really been looked into.

cheers!

[Brian Degger]

Like this? Artist transforms moss with luminous genes
http://juntakita-artworks.blogspot.co.uk/2012/03/light-only-light-light-only-light.html?m=1

On 11 Nov 2015 20:41, "Nico B." <perpetuatet...@gmail.com> wrote:

Has anyone done any tissue cultures or transformations with Spanish moss? looking for a start on how to crow in culture and eventually transform to express some bioluminescent protein that has yet to really been looked into.

cheers!

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[Nathan McCorkle]

On Wed, Nov 11, 2015 at 4:28 PM, Brian Degger <brian....@gmail.com> wrote:
> Like this? Artist transforms moss with luminous genes
> http://juntakita-artworks.blogspot.co.uk/2012/03/light-only-light-light-only-light.html?m=1

"Except for a few species like the Dinoflagellata, which belongs to
both the plant and animal kingdoms, bioluminescence is only found in a
few animal species."

I guess they forgot fungi.

[Brian Degger]

remember, this was written by an artist :)  Its not perfect...and you could always drop him a line. I feel sure he would just make the edit. Or keep it as an artistic statement ;)   

However 

I'm sure the average (non marine biologist) scientist  wouldn't be as informed on pigments used by artists in the middle ages.

Consider

"except for cochineal, which is a product of an insect, pigments only come from plant and mineral products"

he would immediately say what about ....Tyrian Purple  (as would any marine biologist :)

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Brian Degger

twitter: @drbrian

http://makerspace.org.uk

http://transitlab.org

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[Nathan McCorkle]

On Wed, Nov 11, 2015 at 5:14 PM, Brian Degger <brian....@gmail.com> wrote:
> remember, this was written by an artist :) Its not perfect...and you could
> always drop him a line. I feel sure he would just make the edit. Or keep it
> as an artistic statement ;)
>
> However
> I'm sure the average (non marine biologist) scientist wouldn't be as
> informed on pigments used by artists in the middle ages.
>
> Consider
>
> "except for cochineal, which is a product of an insect, pigments only come
> from plant and mineral products"
>
> he would immediately say what about ....Tyrian Purple (as would any marine
> biologist :)

So does that mean painting with blood, or getting blood-stained
clothes are not 'pigmented'? Does that also mean oil is a mineral
(mineral oil is named so because it is extracted from mineral
'containers')?

[Brian Degger]

I knew I shouldn't have hit send.

Keep on Trucking and DIYBio-ing 

Normal service will now resume.

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[Sebastian S Cocioba]

Nico, 

Im thrilled you want to delve into the dark-green arts! I have no experience with moss cept for physcomitrella patens but either 1-2mg/L of 2,4-D or a cocktail of 1mg/L BAP (6-benzylaminopurine) and 0.1mg/L NAA (napthaleneacetic acid) on MS media with gamborg b5 vitamins or with niacin at pH 5.8 is my go to for shot-in-the-dark tissue culture protocols. Replate every two weeks and until callus and then shift to just cytokinin and some giberellic acid to stretch out any stubborn shoots-to-be. Just go ease on the gibrellin since it can induce floral maturation in-vitro and force life cycle changes. Agrobacterium is my goto transformant but now with my shiny new gene gun ill be moving away from my little oncogenic friend and onto the tungsten pastures of biolistics. You can always harass Stan Gelvin at Purdue for his GV3101 agrobacterium tumefaciens strain (LHA4401 is decent but i prefer the more aggressive albeit harder to get rid of GV3101) or source it from your local university plant lab. Someone has to have it if they work with plants and genetics. Best of luck and dont give up!

Sebastian S. Cocioba

CEO & Founder

New York Botanics, LLC

On Nov 11, 2015, at 3:41 PM, Nico B. <perpetuatet...@gmail.com> wrote:

Has anyone done any tissue cultures or transformations with Spanish moss? looking for a start on how to crow in culture and eventually transform to express some bioluminescent protein that has yet to really been looked into.

cheers!

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[Sebastian S Cocioba]

Also be careful since Spanish moss is an invasive species and the EPA and USDA are specifically bothered by release of a plant pest or weedy being. Last thing we want is spanish moss gaining herbicide tolerance. Try positive selection using phosphomanose isomerase from E coli instead. It makes plants able to eat mannose while wild type cant so non-transformed cells survive. You can even design loxP sites and Cre-Lox the selection marker out entirely. Pmi is what golden rice uses too. Herbicides are sooo 90's. :P

Sebastian S. Cocioba

CEO & Founder

New York Botanics, LLC

On Nov 11, 2015, at 3:41 PM, Nico B. <perpetuatet...@gmail.com> wrote:

Has anyone done any tissue cultures or transformations with Spanish moss? looking for a start on how to crow in culture and eventually transform to express some bioluminescent protein that has yet to really been looked into.

cheers!

--

[Yuriy]

You people do know that the term Spanish "moss" is a misnomer. Right?

As for the original poster. Lookup "pineapple transformations". It's the closest commercially viable species thus most researched. Still works with agro.

Bromeliad transformations coughed up next to nothing in search.

[Sebastian S Cocioba]

Saying that culturing spanish moss, where tissue culture has more physiological rather than phylogenic factors to account for, like you would pineapple may not be the best advice. Thats like saying to care for a manatee one should look to literature on elephant husbandry since they are very closely related. Even different cultivars within the same species may have drastically different hormonal responses (annecdotal) so take any literature protocols you find with a grain of salt but do try them out. You never know...you might just get lucky! Although members of the "airplant" family (also bromeliads) may give you some decent leads on micropropagation, as Yuriy stated its a bit scarce post cursory google search especially now that sci-hub is down.

The thin long leaves will be tricky to surface sterilize and may damage those awesome dust-eating tissues on the surface. The high surface area due to said cells will require some dish soap along with bleach to break the tension to get all up in there. Bromeliad seeds suck at germinating so leaf tissue is your best bet...especially for the moss who lays vegetative for most its life.

Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC

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[Marc Dusseiller]

Making things glow... Still seems to be the favourite way to spend time :-)

hackteria.org/wiki/Explorations_in_BioLuminescence

At the moment II enjoy the light of 20kg of rotting wood in my attic. It contains the bioluminescent fungi, Amarillo, found around the house...

Good luck,
M

[Marc Dusseiller]

[Marc Dusseiller]

[Nico B.]

Thanks for all of the responses! Sebastian, I'm adding you're 'shot in the dark' media to my lab book. When i wrote it down on some scrap paper i got that eerie feeling like I'd probably be using that formula quite a bit. I tried with some old media I had at home (Maltose, NAA, BAP, MS and 2,4-D) and tried to surface sterilize with bleach/ no surface sterilization.... all in all I have dead tissues and contaminated tissues. gonna take a bit of trial and error and a lot more reading up but sticking with it! I'll let you know how things come along! Really appreciate the insights from such a practiced wizard!

Also Marc, Thanks so much for the link! I'd never heard of hackteria.

Best,
Nico