For the reason of choosing manual weeding，because Qumai are herbs, spraying the herbicide may cause the persecution of the seedlings of Qumai, so we only choose artificial weeding, and it's not reasonable to use the machine to cut grass, because weeds that just come up is too small, and similar to seedlings. We can only manually use sickle to pull grass and cut grass, causing us a lot of trouble.

He indicated that there were some disadvantages in common herbicides. At present, glyphosate is commonly used, but the effect may be slow after spraying, and it will be inhaled, so it is necessary to plow before planting to prevent glyphosate from destroying the growth of the Qumai.

Accept and expect our results. Weeds grow too fast, there may be weeds after the next rain. Recently, Shangluo has a lot of rain, and weeds grow one after another, which will lead to the waste of a lot of manpower and material resources for weeding, and the growth of the Qumai is also slow. It would be great if there were such a herbicide!

In view of the reason why glyphosate has been gradually replaced，he pointed out that the safety of the use of glyphosate pesticides. A large number of authoritative studies have proved that normal use of glyphosate is very safe, and it is the most important pesticide at present. Most herbicide resistant transgenic crops use glyphosate resistant genes. Therefore, the safety of glyphosate may not be the main safety concern in the pesticide field. We should pay more attention to the problem of herbicide resistance. For example, resistance rather than safety is the main reason that restricts the application of glyphosate;

It is necessary to produce high yield and low cost as much as possible. He said that I think it is meaningful to research and develop the engineering strain of glufosinate-ammonium analogues. However, whether the engineering strain can produce high yield glufosinate-ammonium analogues to achieve the purpose of weeding is the main concern; Second, whether the production cost of the bacterial agent has application potential also needs to be carefully evaluated.

Avoid sub lethal state. ，If the release amount of glufosinate-ammonium analogues analog is not reached, it will not kill weeds, but will make it exist in the environment. Instead, it will be in a sub lethal dose. Increasing environmental selectivity will often lead to the generation of drug resistance. Therefore, it is necessary to note the release amount and conduct actual tests before release.

Improve the synthesis rate of L-form as much as possible. The herbicides we synthesized belong to amino acid derivatives. Research shows that only L-form has this herbicidal effect, and D-form has almost no activity. Therefore, it is significant to improve the synthesis rate of L-form as much as possible in production. The cost can be significantly reduced.

Experimental suggestions. You wanted to be scalable. And I know that the iGEM team from Singapore, two years ago, they made a similar system, also, maybe a blue light induction system for bioproduction, because then you can have a bio reactor full of bacteria, and then whatever you want to induce, you can very easily induce the whole tank, rather than adding a chemical. So this will allow you to grow a lot of cells with a very low amount of glufosinate-ammoniumand and then induce the expression when you needed. And otherwise you're talking about lysis system. And that seems to be a good way to open the cell and get it to leak out. So the only thing I could consider to get to kick things started would be a phage infection, because many phage, they shut down the whole cell, right? So if you want whole cell to start focusing on your glufosinate-ammonium production, then I don't know if there's something that could be done there where you use elements from a phage that is turning off everything else, but I don't know how much you want that either, because you need, you probably need in multiple metabolites and enzymes from the cell to support the production. It's something you could consider, right? Some phages, like T seven, it has 11 minutes in the cell, right? So has 11 minutes completely changed the time metabolism, destroy the cell and then get out. So it's, it's very efficient at that. So it could be something you could look at a little bit like your lysis.

Discuss uncertainty about bacterial release， It depends on where in the world you go. So in certain parts of the world, you will not be allowed to spread GMO like that, not yet , but other areas you could. I don't know how how nature reacts to huge amounts of Ecoli. You could say we're using animal and and also human fertilizer. Have used that in agriculture for many years, and that is a lot of that is Ecoli, right? So I don't think it has to be a problem. After words, it's a bacterium, but it's a pretty clean bacteria from your spreading here. If you talk resistance, so you should probably get all antibiotic resistance markers out of the bacterium before you spread it so it can outspread antibiotic resistance, right? I think that will be necessary to do. And you may also have to do something so it can't do conjugation, can't do horizontal gene transfer. It should be possible to do something there to protect it a bit, but you can never be sure if your bacteria miss alive on the field, even briefly. Then you could also have bacterial phages that can go in, grab DNA and move on, right? So try to avoid problematic elements. And then you're talking resistance to the growth of weeds. We don't really know the mechanism, but I assume that there will be resistant at some point. But that was case, so I don't think you can avoid that. It would be a test and see how long can it last, and how much those is there. So we know from other treatments, antibiotics, e.g., that if you give a very low dose for very long time, then you give the the pathogenic bacterium quite a good window to become resistant, right? Where if you give a higher dose, and you just give it once, then you can kill them. So the same could be true for your field here, that if you give too little of your compound, then the weeds have a better chance of becoming resistance. If you want to use it, you should really use it. But it's difficult to convince the farmer to use a lot, I guess.

a cell-free system . I think that's a really good consideration to take in. You can see the spirit of iGEM is very much to pack everything into machine. But cell free is definitely not a bad idea to think about. Or simply just say, you're producing this in a bio reactor, and then you harvest your herbicide, and then you, and then you spread the herbicide specifically, right? And you don't have to purify it very finely. You just have to treat it in a way where you think, now there's no live bacteria and almost no genetic material, so you're not really exposing nature to that. And I think that is a very viable, may be a little academically, a little less exciting, but a very viable solution. So it's actually a matter of bio production rather than a superbuck that can kill the weeds and hang out on the field. So I think both are really good to have in mind.

The aspects we need to pay more attention. I think it can have very great significance if it's acceptable to use, if it's safe to use, and it actually has a beneficial effect. And we are spending a lot of energy on making artificial fertilizers. if your bacteria can be used as fertilizers, then you can really help, especially small farming, but maybe big farm as well, if it is safe. The safety is, of course, the big issue right here, and that's probably where you, in your previous question, had the answer with the cell free. And I think you may be able to get around some of it, but surely some of the bacteria will get around it and start growing, right? But that's not necessary a problem either.

Your expectations for the future development of synthetic biology Synthetic biology of bio economy is predicted to be one of the big pillars in the 4th Industrial Revolution. I think the more exciting bit is probably the the self therapy that is increasing. I would characterize that as synthetic biology as well, where you can reprogram cells to do something practical. And we're not just talking fixing genetic diseases. Car-T therapy is probably the most famous, or most useful examples so far, but I think we're gonna see a lot of that. When we take cells, we reprogram them to do something functional with a logical circuit, and then we sent them into the body. We have to be careful. They are dangerous. There's a lot of considerations. But it's a very appealing and promising area.