We are making Glowing Plants using Synthetic Biology and hope to one day replace streetlights with glowing trees. Ask us Anything!

Do you have any idea how much astronomers are going to hate you? At least you can turn a streetlight off.

Actually there are several ideas we have for how to reduce light pollution. The most promising is to only express light in the bottom of the plant's leaf, so it's only reflected down.

There are also ways to turn the light on and off using different promoters. The most fun is one which responds to touch just like in avatar!

Do you think it would be possible for you do another crowd funding project for grass in the future where it will light up on contact?

Maybe :)

That sounds really interesting! Could you maybe point me to some research on that promoter if you can? I'd love to learn some about that and what has been done with it if its out there. Thanks.

Here's one (there are many): http://jxb.oxfordjournals.org/content/60/1/43.long

How bright can we really expect these plants to be compared to a regular street light or even a 60 watt bulb?

Initially, not very bright. We are aiming for a plant which glows like glow in the dark paint. Longer term however when we really get into the metabolic engineering I think a plant could be as bright as a bulb... the question is how big a plant we need. I'd hope for a Christmas tree sized plant to be able to replace a 100 watt bulb.

Could you give us details on how bright it is?

We haven't made it yet, the kickstarter campaign is to raise funds to do so. Researchers at SUNY were able to make a plant which visibly glowed after you were a few minutes in a dark room and we are building on that work. We go into detail on the techniques we are using to do that here: http://www.kickstarter.com/projects/antonyevans/glowing-plants-natural-lighting-with-no-electricit/posts/468578

For inspiration on how bright bioluminescence can go we look to the 2010 Cambridge iGem team who were able to read books from their glowy bacteria, more information here: http://2010.igem.org/Team:Cambridge

How will glowing plants effect nocturnal animals?

That's a good question. Chances are our glowing plants will be much less bright than a full moon. And the brighter we can make them, the less likely they will be able to compete in the wild. So the concept of brightly glowing plants spreading rampant and causing light pollution and affecting insect populations is highly unrealistic.

Insects would have as much trouble with our plants as with the actual street lights the glowing plants are replacing. Probably far less, because the light producing surface would be much more spread out than you'd get with a light bulb.

Is gene escape a possibility using your methodology? If you have glowing Arabidopsis, would you expect there to be some escape into the wild and crossing with non-glowing populations?

The glowing effect uses energy - it takes a burden on the plant metabolism and sustainability. Even if this would "escape" into the wild (like countless genetic modified bacteria and yeast did during the 40 years we're been doing genetic engineering) - it will not prosper in nature.

1) Where does it get it's energy? Does it absorb it from the sun during the day? That doesn't seem to be a hindrance to it's spreading.

2) Would it attract moths?

It uses ATP which is what all living things use for energy - the plant makes it from Photosynthesis. The point is that the glowing effect imposes a metabolic cost on the plant, which means plants without the glow effect will out compete it - evolution in action! we are also exploring some additional methods of keeping the plant even more dependent on human care

Hey, I am a biochemist, can you reveal any of the specific biochemistry you're doing here? As in how you are producing the glowing. I've only ever done this with plasmids, but I suppose you could do it a better way.

Hi! as a fellow (past) biochemist - thanks for your question!

For Arabidopsis / agrobacterium transformation we're going to insert our synthetic DNA design into T-DNA plasmid. Once we got the final design we will use the gene gun method. The design will include all the genes from the bacterial LUX operon (codon optimized for plants expression). We will try different plant promoters and genes combinations to improve the glow effect.

I guess you have to disarm the T-DNA plasmid so that all your plants don't die as well then. How are you causing the uptake of T-DNA plasmids? As I remember agrobacterium require the release of phenolic compounds from an injured plant.

Fortunately, plant molecular biologists have been working with agrobacterium since the late-70's early 80's (we are humbly standing on the shoulders of giants here). A lot of disarmed T-DNA plasmids exist (just as there are plasmids for E. coli protein expression). While there are a couple of ways to get agro to take up T-DNA plasmids, we've been working with the cold-snap method - which gives us an excuse to play with dry ice. Yes, acetosyringone does help transformation but it doesn't seem you have to always add it externally when doing experiments.

That sounds awesome! If you guys weren't so far away I'd love to get involved.

One of the reasons for being open and transparent is to give people further away a chance to get involved and contribute in whatever way they can. Don't let distance from us stop you!

I'd be happy to help, as a student I have a lot of free time during the summer but not a lot of money. Aside from just backing on kickstarter, how can I get involved?

Look out for our $500k stretch goal - it's going to enable anyone to participate in the project.

We are using the disarmed version of Agrobacterium... specifically GV3101 strain

Will this be brought to third world countries? Ones with a weak electronical infrastructure.

I think this could be one of the most trans-formative applications of the technology. Lighting has been shown to have many benefits to reducing crime and improving quality of life and there are large parts of the world which lack night lighting (just look at a photo of Africa at night). Tropical countries, with ample sunlight and equal day night lengths, are ideally suited for this technology so I think it could be something which finds acceptance there before more developed nations.

How much maintenance do these plants need? What will happens over time if they are in rural, deserted areas?

The plants we are initially using, for the kickstarter campaign, need a lot of maintenance. They need quite particular growth conditions, eg not too much direct light, not too hot, cold winters (to germinate seeds). In rural areas they are likely to not survive very well - this was one of the reasons we decided to use this plant.

What sort of regulatory challenges do you expect to face? I know big companies spend a lot of time going through the deregulation process of transgenic plants.

Also, since this is more than simply expressing GFP and making the plant fluoresce under a black light, what kind of effects does the glowing have on the growth of the plants? As I understand usually plants that actually glow express the enzyme and are watered with the substrate instead of producing both. And as for glowing trees, is there any proof of concept that the trees could emit that much light on their own? That seems very...interesting.

Also, I'm sure you are aware of the controversy surrounding genetically engineered plants. Have you guys encountered any opposition? I hope that plants like this would enable to see and hold GE plants in their hands and realize they aren't the Frankenstein monsters some people say they are.

Thanks, and best of luck.

Controversy: A Canadian NGO has asked us to cancel the project. Here's their request: https://dl.dropboxusercontent.com/u/17237718/NGOs/letter%20to%20Evans%20et%20al%202%20May%202013.docx

And you can read our official response here: https://dl.dropboxusercontent.com/u/17237718/NGOs/Response%20to%20ETC%20Group%206%20May%202013.pdf

Gene expression: We are actually putting the full luciferin pathway into the plant as well, so it won't need the addition of a substrate to glow. Longer term, getting really bright plants, is going to take some more significant engineering so all we have for now are some theoretical calculations but it does look feasible. The big unknowns are how efficient the light production will be (fireflies are almost 100% efficient) and how much of the plants metabolism we can devote to light production without stinting the plants growth.

Regulatory challenges: There are three federal agencies which regulate Genetically Modified organisms in the US, each with a different remit for public safety:

USDA regulates plant and agriculture impact through APHIS and are the most relevant for our project. We've been in touch with them to understand and address their main concerns which are mainly related to the introduction of potential plant pests. After more than 15 years working with genetically engineered crops they have established a set of guidelines for what needs additional testing, and what doesn't. So long as we meet all their requirements we can safely release the plant. One of their inputs was that we should use the gene-gun technique to transform our plants, instead of Agrobacterium. EPA regulates new uses of pesticides - many GMO's introduce pesticide or herbicide resistance to their plants (either as a selection agent or as an intended outcome). We have elected not to do this, as we can use the glowing effect as a marker, so will not need to go through their testing procedures. FDA regulates food and feedstock implications and requires extensive testing to make sure the product is safe if this is the case. Because our plant is strictly ornamental and not for consumption by animals or humans we do not have to go through this testing.

Great work you guys are doing there! I'd have 3 questions for you in relation to the new interface in between synbio and the general public kickstarter is offering: 1) Could you share some data about who your backers are? Scientists? Hobbyists? Techies? What age range? 2) You did a great job at science popularization - what principles or tricks did you follow? Did you work with journalists or marketing experts for exemple? 3) How do you fix the price to a thing that does not exist yet and where there's no market to start with? Thanks!!

Thanks for your support and interest in the project 1) We don't have a lot of data about who the backers are, but based on the traffic and blogs I would say it's a mix of scientists, DIY Bio people, techies and futurists. A mix of genders and very broad range of ages. We'll ask some of these questions in the follow up survey and will share for others interested in the campaign. 2) The most important thing we did was user research. We showed the landing page to over 100 people before launch and significantly changed it. A lot of scientists have critizised us for not going into the science enough on the page, but this was something which specifically came out of this process. It's a hard job keeping things simple enough to not scare people off, and yet more complex for people who want to do their due diligence to actually do it. If I did it again I'd create a separate site which went deeply into the science and then have a link to that page from the site. 3) Pricing was really hard. Initially we thought to charge $100 for the seeds, but the user feedback told us it was too high. People told us they would pay $25-30 for the seeds, so we went a big higher as we figured they would pay a touch more than they said they would. It's an art not a science, and we had to trade-off the cost of delivery with having enough spare cash to fund the science.

Thanks for the reply! About research before launch, it makes me think about the lean startup concept ;) I have two follow-up questions: 1) For this pre-launch research phase, how did you probe people (for the design, content and pricing)? Any specific software / platform or tricks there? 2) Being endorsed was probably very important for you. How did you get all of these big names? Is there money incentive for them or its just scientific interest? PS: I'll be writing a small blog post here if that's ok about kickstarter as a new platform Synthetic Biology. It's a new Synbio Consulting startup I'm launching ;)

1) I talked about it constantly to everyone I met for six months. Thanks to all my friends for being so understanding. We used every platform you can imagine to get feedback: if you walked into my house you had to sit down in front of hte video, we emailed it to mailing lists (eg biocurious) and I emailed it to friends, often three times! 2) There is no money incentive to any of our advisers. Omri has been building relationships with many of them for a long time, and that helped, but it also helped that we are trying to do this the right way for the long term interests of the syn bio community and to raise awareness about this tech. I can't say we got that right 100% of the time, but we are very aware of the precedent we are setting in doing this and have really engaged and focused on the complex legal/social/ethical issues raised. 3) Sure write a blog post, I'd be happy to discuss that with you offline... my contact details are on the kickstarter page.

Do you think they will make glowing insects if the bugs eat them?

No, the glowing effect comes from the living cells of the plant. Once a bug eats the plant it will kill those cells and they won't produce the proteins anymore. Actually you have probably eaten these genes in very small quantities, if you have eaten squid, as many sea creatures eat the bacteria from which the genes come from. The plants are certainly not food for humans or animals though!

Hi, i'd like to know if the plant would use the ATP from its photosynthesis to light up, or if it uses a different process. And does the reaction of the luciferase can interfere with other reactions in the plant? Finally, what makes it harder to create a bioluminescent tree instead of a small plant? Thanks!!!

Yes, it uses ATP. This is the metabolic pathway: https://dl.dropboxusercontent.com/u/17237718/Reward%20images/bacterial%20lux.jpg

The biggest challenge with a tree is the length of time it takes to grow, which is essentially the limiting factor in each experiment. I think to make this work in trees we will need to develop better simulation technology - lots of teams are working on this so hopefully it will be developed.

Along the same lines. With small young plants like arabidopsis almost all of the tissue is primary (green). Do you think it would be possible in future plants (like Christmas Trees) to get older tissues, non-green tissue, to glow? Bark, what about Bark... are you focusing on this at all?

Bark is dead cells, so probably we can't get that to glow (we need ATP and recylcing enzymes)

Thanks for doing this AMA, I think what you guys are doing is amazing and I'm excited to be getting seeds for a glowing plant next year.

So for my question, about how long do you think it will be until you can walk into a home improvement store such as Lowe's or Home Depot and purchase glowing flowers, trees, bushes, etc. alongside non-glowing plants?

We've actually been approached by several distributors interested in helping us get future plants into stores. The main thing we will need to work on are bigger plants and getting a brighter effect.

I've already asked you guys a bajillion questions on kickstarter, Antony has done an exemplary job answered all questions on the kickstarter page.

Are you guys going to release the exact growing methods and conditions (lighting, brands of nutrients, etc) that you use in the lab to grow these?

Also, I think the rose stretch goal would probably be best suited to be white, as it would probably give off a nicer glow than if it were red or some other color.

Hope this launches your campaign on kickstarter on another level. Any chance you could reveal your next stretch goal on this AMA?

Maybe we can reveal that... let me discuss with the team!

Awesome. Can't wait!

Here's a hint... it's at $500k and it's not a new plant...

A DVD documentary on the glowing plant, start to finish?

It's more a way for backers to get involved with the process :)

We are kind of going to do that anyway. It won't be a DVD but we will be blogging all our work and sharing videos online, plus of course we will have the book.

What exactly happens if you accidentally kill a glowing plant? Will it be easily compostable?

Can the plant survive if you put it under a shade? Because most places that need to be illuminated are already very dark.

And finally, can you turn the plant off or is it glowing during their entire lifetime?

If you kill a plant it will behave just like any other plant. the plant we are using in this project actually prefers to grow in the shade, but obviously other plants will be more efficient at converting sunlight into energy so for really bright plants they will probably need direct sunlight.

For now our plants will be on at night all the time, but we are exploring ideas for the future on how to turn them on and off.

Is this idea going to be patented and restricted for use by others?

We are going to patent our work, but only for defensive purposes. All of the output from this project will be open source and in the public domain. In fact if you download the Genome Compiler software you can see some of our designs: http://www.genomecompiler.com Someone already order the DNA for one of these!

First off, I want to say that this idea sounds amazing. I feel like apocalyptic fiction written in the near future may have to account for glowing trees still existing in a world without electricity. :)

My questions (apologizes if they have already been answered): will these plants glow for their entire lives?

Will they be able to spread on their own, resulting in glowing plants all over the place, or fields of glowing plants?

Yes they will glow for their whole lives (but only when alive!), and they will be able to reproduce. Unlike monsanto we think if you own seeds you should own the offspring. That said there is low risk that they will spread uncontrollably - this is not an invasive species and the glowing effect will impose a significant metabolic cost on the plant reducing it's ability to survive in the wild without human care.

Where do you think Synthetic Biology will be in five years time?

I want to see a world where small teams of entrepreneurs can create applications solving major world problems, eg the low cost malaria drug. To help stimulate this we are going to write a policy document sharing how we have approached the legal, social and ethical issues related to this kind of work and hold a conference in the fall.

What other projects are you looking into? I saw that you made glowing bacteria before this plant, so what is the next project if any?

Our main focus for now is the improvement of the brightness from the plant and moving to bigger plants which can produce more lumens.

We hope to inspire people with this technology and get more people working on synthetic biology projects. There are so many applications in energy, agriculture and medicine.

I can just see this thing going horribly wrong and all the sudden we have entire forests glowing and insane light pollution. The only safe-guard you have to that is that other plants should out-compete these ones? What about places where there aren't that many other plants to compete with? And how do you make sure that other plants can out-compete these?

Also, what about these plants evolving into not being so susceptible to being taken over and dieing?

The biological world is brutally competitive. Plant's don't really decide what genes to keep and what not to keep, evolution works to optimize the plants survival. If there are no other plants in a location it's probably because it's a tough place for plants to live (eg salty, no water) - our plants would have no advantage over other plants there. The most likely way our plant will evolve is to lose the glow effect, not get stronger in another way, as it confers no selection advantage. We will have to look after the plants to make sure they keep the effect. This is actually one of the biggest problems with scaling up bacterial production of protein production as the bacteria keep evolving to not produce the protein (plants are a bit easier as they live longer and don't evolve so rapidly)

Any plans to ship the seeds out of the US?

Is there much opposition from luddites?

We'd love to ship them abroad, but regulations (especially in Europe) for this kind of product are complex and vary widely. It's probably not cost effective to ship outside the US for now as the cost of regulatory compliance will be bigger than the market.

There's been a little opposition from environmental groups, we did expect this. You can read more here: https://dl.dropboxusercontent.com/u/17237718/NGOs/letter%20to%20Evans%20et%20al%202%20May%202013.docx

And you can read our official response here: https://dl.dropboxusercontent.com/u/17237718/NGOs/Response%20to%20ETC%20Group%206%20May%202013.pdf

If you create a GMO glowing tree, will it naturally attract unobtainium underneath it?

Yes - and we use blue avatars to extract it!

No question, I just want to tell you that you guys are lumping awesome!

Thank you!

Why don't you have any pictures of the actual plants available on your Kickstarter page? Have you actually made working glowing plants yet? It didn't seem like it when I saw your campaign page.

We don't have working glowing plants yet which is why we can't show any pictures of them. We are using kickstarter to raise the funds to synthesize the DNA and make the plants. We have designed the sequences, and we have tested the transformation protocols in the lab.

So if the plants are storing the light energy to be released in bioluminescence then that energy Is not being used for photosynthesis and therefore growth, what would this have on effect the rate of growth? And how long would you propose it would take to grow a tree feasible of replacing a street lamp?

Plant's clearly have an abundance of energy - imagine how much a potato stores as starch - so they should have enough for light. One of the big unknowns for how bright we can get this is how much of that energy we can divert without affecting the plant, nobody knows the answer to this yet.

The big problem with trees are that they take a long time to grow, so we can't easily do experiments on them. We probably need better simulation tools to make this happen but people are working on this (see collaboration between Genome Compiler and Autodesk's Project Cyborg team).

Fair enough, which plant species are you using to test on? And which tree species would you ideally be using?

WE are using Arabidopsis for the first plant. For trees we will probably use something like the fast growing trees JBEI are working on.

Is it possible to genetically alter plants to take in carbon monoxide instead of dioxide? I had an idea to do that and plant a shit ton of them in the empty parts of the four-leaf-clover parts pf highways. Eventually unfuck the atmosphere. Bonus: alter corn to do that, lead to more ethanol.

In theory that could be possible... why don't you try to do it?

I would like to make the request that if and when you get a full compound lab for more development research, could you devote one wing to biome where the public could tour and see how your developments are progressing and to give people a grasp of the day in the future where this may be more common place in the normal day suburbs.

That's something we are going to do. We will be in touch in the summer with backers inviting them to come and visit the lab.

Which goal are you trying to achieve more: using less energy or putting more oxygen into the atmosphere?

Reducing dependency on electricity for lighting needs.

Do you have any idea where these will be located first if they ever gain popularity/work?

Most likely trees which replace streetlamps will start in developing countries.

As a recent college graduate (molecular biology), how can I get involved in synthetic biology? What skills and experience should I develop to ultimately work on a project such as yours?

I think the best experience is doing this stuff in the lab. We will be taking on some interns to work on the project with us, if this is interesting email me (contact details are on the kickstarter page).

This is honestly the coolest thing ever, and I hope that streets are lit up with glowing plants in my life time. Unfortunately, I don't have much money, or else I'd absolutely donate.

Do you think that the first glowing plant "street lamp" will happen within the next decade?

That's probably the right order of magnitude of time... I would say 5-10 years at least.

I looked at your kickstarter before and I was very interested, except the only thing stopping me from providing a donation was that you had no actual pics/videos of the actual plant itself apart from that one iconic picture. Do you have any actual pictures of the plants? Or maybe some video showing what the plants look like when illuminated?

We don't have the plants yet, we are still raising money to make them.

I think that the LUX flowers are pretty cool but I was wondering if you've done any estimation of the cost/carbon emissions of maintaining these plants over street lights? I can imagine that at least they will require semi-regular weeding

Big trees don't require any fertilizer/weeding in existing streets so we should be OK.

At what point do you think we will be able to make people glow?

Bioluminescent tattoos would be awesome.

I would like one of those, but I think the FDA is going to cause some challenges for doing that!!

Do you think this is the future? Glow in the dark forest houses?

I can only imagine the lights going out "Damn! Forgot to water my lights!"

Oh! that sparks another question, if the plants die do the lights die?

Ah what the heck one more, does this work with moss?

Growing houses is an interesting idea...

If the plant dies the lighting effect will die as well as the light effect needs ATP to glow.

Yes, in theory it would work with moss. More research on moss would be needed to understand the metabolic pathways first.

This is so exciting! Imagine a glowing forest?! The plants still function properly in terms of converting Co2 to o2 right?

Yes!

Will they grow after they die? What exactly will glow? (as in, will the whole plant glow, or the leafs?)

When they die, the glowing will stop. For now the whole plant will glow, but in future we can test different promoters for different parts of the plant. One effect which would be cool in the future is different colors in different parts of the plants... lots more research needed to make that happen though!