Team:Marburg/Human Practices/Integrated/KWS

KWS

Overview

KWS is an internationally known seed company whose Headquarters are located in Einbeck, Germany. They improve genetic potential through outstanding research, breeding programs and supply the farmers with top-quality seeds. The company's various projects include nitrogen-efficient corn, a new generation of herbicide-tolerant sugarbeet, hybrid potato seeds and maximizing photosynthetic efficiency in corn. In addition, KWS is involved with trait detection from the air and helps with the Crops of the Future initiative.
We conducted our interview with Klaus Schmidt. He is the Group Lead in Plant Cell & Transformation Technology, which means that he produces transgenic plants for research purposes to better understand them. This makes Klaus Schmidt a suitable contact person for our project to discuss various problems, such as the transformation of plants.
When we asked him what the big bottleneck in plant transformation is, he answered that from his point of view regeneration is the biggest problem, which means that a transformed cell is regenerated back into a complete plant. Depending on the plant, this is very difficult, or it also depends on the genome. Another bottleneck, especially in chloroplast biotechnology, are the technical hurdles to get homoplasticity plants.


[...] it's really difficult [..] to get really a homoplastomic plant. It requires selection, so you have to introduce an antibiotic resistant marker and normally this is done [..] with spectinomycin resistance. And if you want to bring it to the market, antibiotic resistance is a no go. So, if you could do it without an antibiotic resistance marker or using an alternative marker system, this will be maybe a way forward.

(Klaus Schmidt)

However, there is no plastid transformation method for corn. Besides these issues, the dream of chloroplast transformation projects has not died in KWS, some small projects are pursued in this direction. This again clearly shows how difficult it is to work with transplastomic plants, both in industry and in research.
Our project therefore could substantially assist with the necessary basic research, regardless of the current legal regulations. According to Klaus Schmidt, our cell-free system could, for example, be used to pretest different promoters and later screen for the same mutations in seedlings in order to increase a desired specific protein concentration. In regard to this, the measurements always depend on the type of output, so a protein concentration can be measured using GFP in the CFS, but the sugar content must be tested in the field. A herbicide resistance could also be pretested in CFS by modifying the responsible enzyme.
In addition, research with chloroplasts would offer further advantages. These include high protein expression, herbicide tolerance and insect tolerance. For example, bt-toxins for insect resistance could be synthesised in the plastids, which would prevent them from being toxic to the plant itself, since they are encapsulated in compartments of the plant.

During the interview, it also became clear to us again and again how strict the regulations in Europe are in contrast to other continents. Many projects are simply not possible here, because either the end product might not be saleable or even the type of production leads to problems, although the end result is often indistinguishable. This makes the work in synthetic plant biology much more difficult.
Besides the technical hurdles, we also received some interesting insights regarding the legal hurdles. Regarding the question of why GMOs are so strictly regulated and why public acceptance is so low, we got an interesting answer.

It’s simply about fear. GMO seems to be something different and people might say: “I will not eat something different. [...”] Coming back to potatoes, for example, in Germany potatoes have to be yellow, in other parts of the world, they need to be white. There are these cultural differences. Also for corn, the same in Africa, corn/maize has to be white, otherwise people won’t eat it. People simply don’t like it. It’s not understandable, but that’s how it is.

(Klaus Schmidt)

This makes the situation a little amusing, but explains quite well that people do not really trust a slightly different food and that is perhaps why the regulations are so strict.
During the interview, other very interesting points came up, but in summary Klaus Schmidt said that there is a lot of potential in synthetic plant biology. For example, one could compare it with covid vaccinations. As soon as a need arises, the potential can be really exploited and synthetic biology could become really important. To realise this, the appropriate funds would also have to be made available. Especially in the next few decades, the need for synthetic plant biology could increase greatly in different areas.
It was also described in the interview that in the next decade we will need much better models for interaction in plants, like a kind of AI that models the outcome before the field tests can be done. Our cell-free systems can also help in developing the models by making it much easier to characterise the plant genes. In conclusion, we are helping to open up the great genetic potential of plants in biology.

Insights

Besides all the interesting insights into the industry, we also took away some ideas for our project. On the one hand, we want to measure the length of the cell-free reaction in the long term, so that we can also test more complex metabolic pathways.

Measuring GFP, you need to produce the protein. I don't know how long it will take; a few hours in order to get sufficient protein maybe. So for more complex scenarios at the end [...] to use the cell-free lysate for measuring biochemical compounds. I don't know. So, can you use the cell-free lysate for a few days, few weeks?

(Klaus Schmidt)

Unfortunately, due to time constraints, we could no longer test this precisely, as we wanted to use optimized CFS for the measurements. A non-optimized system also has a relatively low output, which makes half-hourly measurements over a longer period of time difficult. Furthermore, the lack of certain chemicals can also lead to premature termination of the cell-free reaction. In addition, we could optimize the reaction mixture to increase the duration of protein expression and prototype even more complex metabolic pathways.
Another possibility is an increase via semi-continuous measures. Here, the cell-free system is connected to other chemicals via a semipermeable membrane. If the cell-free system needs, for example, more magnesium or creatine phosphate, then these are automatically added back to the cell-free system through a concentration gradient.
These ideas leave some room for experiments in the future. Another exciting idea, which also builds somewhat on the idea from the interview with Corteva, is to test other selection markers that are not antibiotic resistances.

The testing of the different antibiotics should also serve as a proof of concept to show that different selection markers can be tested in our CFS. However, this also offers exciting experiments after the iGEM project.