Team:ZJU-China/Demonstrate
Demonstrate
Overview
The general functions of our project:
Detecting the phytopathogens or the unhealthy situation of the plant by Trichoderma atroviride or our device.
Signal transduction and amplification
Expression of downstream genes
When our engineered Trichoderma atroviride meets phytopathogens, (take Phytophthora nicotianae as an example), some of report genes will be activated and give warning to our device.
How we prove it?
☑︎Cloned the ech42 promoter (the promoter can be elicited when Trichoderma atroviride meets phytopathogens) from Trichoderma atroviride and performed confrontational coculture to test its phytopathogen sensitivity.
三张图
Our device detects the change of VOC(Volatile Organic Compounds) released by plants and estimate whether our plants are infected.
How to prove it?
☑︎ We have constructed a classification model which can tell the health situation from the VOC they released.More details(方哥分类模型的超链接）
Once the device can transmit the order to our engineered Trichoderma atroviride with chemical or electromagnetic signals.
Chemical signals：
☑︎ Cloned the phlABCD cluster and carried out the bio-synthesis of DAPG in E.coli.
☑︎ Constructed the plasmids for DAPG bio-synthesis in Saccharomyces cerevisiae and Trichoderma atroviride.
☑︎ Constructed pho promoter and expressed phlF repressor.
Future work
☐︎ Test pho-phlF system with DAPG
☐︎ Tested the function of phlABCD in Saccharomyces cerevisiae and Trichoderma atroviride and detect the bio-synthesis of DAPG.
Electromagnetic signals：
☑︎ Expressed TRPV-Ferritin in Saccharomyces cerevisiae and tested the its function with heat shock and capsaicin.
☑︎ Constructed Pcdre-mRFP in Saccharomyces cerevisiae and measured the relative intracellular calcium content needed to activate CDRE promoter.
☑︎ Proved that the calcium influx induced by TRPV1 is strong enough to activate CDRE promoter.
Future work
☐︎ Test TRPV1-Ferritin system with medium radio frequencies in Saccharomyces cerevisiae.
☐︎ Construct TRPV1-Ferritin-CDRE system in Saccharomyces cerevisiae and Trichoderma atroviride.
Once our Trichoderma atroviride has received the signal, the downstream gene will be expressed.
How to prove it?
☑︎ We managed to express a special Serine protases in Saccharomyces cerevisiae and tested its activity.
Future work
☐︎ Express this serine protase in T.atroviride.
☐︎ Search and express more downstream genes to equip our T.atroviride with more functions.
