Proof of Concept | PuiChing_Macau

Figure 1. The flow chart of our proof of concept.

To give proof to the efficacy of our concept, our team has done the following experiments.

We transformed the Green Fluorescent Protein (BBa_J364000) to ensure that E.coli can grow normally in the hydroponic system to guarantee the effectiveness of our plasmids. The growth of E.coli in hydroponic water is evidently better than in pure water.

Figure 2. The plate of GFP BL21 E.coli under UV light.

Figure 3. The GFP emission rate of the sfGFP.

To validate that our pH shooting system is able to react to the pH levels changes in the growth medium and neutralize the environment, we tested the system at different pH levels. In all of our tests, the pH converges to about pH 7.

Finally, we monitored the pH levels of the our hydroponic solutions with automated pH sensors over one week. The experiment setup is shown in Figure 4, the result is shown in Figure 5.

Figure 4. Monitoring the pH values of hydroponic solution.

Figure 5. Comparison of glsA, pH shooting, and control under an acidic environment of a hydroponic system with initial pH value is 5.2.

Figure 6. The pH changes in different initial pH environments of the genetic pH shooting system.

To test the inhibitory effect on ABA of our E.coli, we compared the speed of germination of seeds with or without our E.coli. To further enhance our differentiation, we added ABA to some groups. Our results suggest that the seeds with our E.coli germinate faster than those without.

Figure 7. Comparison of the germination of soybeans with and without added ABA, and added with either NSP4-T2R4 transformed E. coli, PYL8 transformed E. coli, empty pET11a vector transformed E. Coli, or LB medium.

Figure 9. The hormone inhibiting test, PYL8 and NSP4-T2R4 constructs.

With the experiments above, we manage to prove the effectiveness of our E.coli in a hydroponic system. However, the results are not perfect. We still need to stabilize our pH shooting system and test for the correct amount of E.coli needed for optimal germination speed.