Team:TPHS San Diego
Overview
Fungal infections have detrimental impacts on agriculture and the environment by decreasing crop yield. In particular, Fusarium Wilt, a fungal disease caused by Fusarium oxysporum, affects a wide variety of crops that make up global food supply. Current treatments such as cultivation are costly, and fungal pathogens can acquire resistance to such treatments. To combat these challenges, our team will be targeting the breakdown of chitin, the primary component of all fungal cell walls. Chitin provides rigidity in fungal species and acts as a similar constituent found in exoskeletons of various organisms like arthropods and insects. Our proposed mechanism for the antifungal treatment is to degrade chitin through the use of chitinase, an enzyme designed for this purpose. Various chitinase isozymes act upon the varying chitin structures in different organisms, so chitinases LbCHI31 and LbCHI32 were selected as the anti-fungal agents against Fusarium oxysporum.
Procedural Outline
The goal of this project was to engineer a bacterial strain capable of secreting viable chitinase to attack the cellular stability and structures of Fusarium oxysporum, thereby inducing structural collapse of the fungal cell wall. A plasmid containing genes encoding for LbCHI31 production and secretion were used to transform bacteria using standard heatshock and recovery protocols. A Western blot mechanism was used for expression of LbCHI31 and LbCHI32. The respective plasmids were tested on chitin to prove effectiveness.
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