Team:CAU China/Proof of Concept

In our whole experiment, we evaluated dsRNA and Metarhizium acridum effect to locust.

dsRNA experiment

We first attained our target genes specific designed dsRNA sequence and fused it with T7 promoter. In order to measure its RNAi effect to locust, we use RNAi expression kit producing dsRNA. Then we injected dsRNA solution with 0.8~2μg dsRNA per locust and injected DEPC water into control group. Here is our result:

The CHS injection group LT50 is 4.0 days, DEPC is 9.0 days and WT group is 10.1 days. By using t-test, its t-test value is 0.025656488, which has significant difference with DEPC control group which means CHS dsRNA worked well on RNAi interference.

We also made ATPase and TSP dsRNA injection experiment. However, from the plot above, even though the ATPase had the same effect with the CHS group, the DEPC (control) group also has a relatively high fatal rate. However, from the searched reference, ATPase dsRNA and TSP dsRNA also can result in high fatal rate (approximately 100%).

Metarhizium acridum effect

Besides, we also employed M. acridum in our project. To evaluate its virulence ability to locust, we made an experiment of measuring locust mortality. In our experiment, we divided locust into two groups, one is control group which were injected with cotton seed oil, the other is experimental group that was injected with M. acridum spores in cotton seed solution. We’ll observe locust survival rate everyday and count its death. Here is our result.

As the plot above shows that the control group LT50 is 17.2 days and M. acridum group is 7.3 days. The t-test result is 0.076, which is a little higher than 0.05. We can regard that M. acridum has infection on the locust but not so significantly.
To test that the death of the M. acridum group is resulted by the virulence of M. acridum, we did locust anatomy experiment. We cut locust wings and observed its pattern. Comparing with control group locust wings, we found that experiment group locust wings has filamentous unknown things covered.

On the left is the control group locust wing while on the right is experimental locust wing. We can assure that Metarhizium acridum has affect on locust living.

Optogenetic automated control system

In our Optogenetic automated control system, we made a relationship between OD620 to our surveillance sensor value. With this plot, we can observe the yeast growth condition in real time. When the sensor value reaches the threshold, our system will transform from yeast growth into dsRNA synthesis.