Creating Mutants Using CRISPR Technology: Investigating Terpene Gene Function in Medicago truncatula

Presenter

Emma Marilyn Meade

Campus

UMass Amherst

Sponsor

Sibongile Mafu, Department of Biochemistry and Molecular Biology, UMass Amherst

Schedule

Session 1, 10:30 AM - 11:15 AM [Schedule by Time][Poster Grid for Time/Location]

Location

Poster Board A64, Campus Center Auditorium, Row 4 (A61-A80) [Poster Location Map]

Abstract

Terpenes are a diverse group of natural products synthesized by plants. Over time, plants have evolved to make these compounds in order to meet specific needs for better adaptation to their environments. Terpenes have a wide range of functions spanning from the plant's defense to attraction and communication with other plants and animals, providing a crucial role in ecological interactions. Our study focuses on terpenes in the model legume Medicago truncatula (Barrel clover), which are used for the plant’s defense. Terpene synthase (TPS) cyclize carbon precursors to make chemically diverse terpene scaffolds. In our study, the genes being targeted produce terpenes with roles in plant defense against herbivores. In order to study the biological role of MtTPS1, MtTPS2, MtTPS3, MtTPS4, and MtTPS15, CRISPR/Cas9 technology was used to “knock out” the TPS genes. The designed CRISPR construct was delivered first by electroporation into Agrobacterium and then into the plants via careful tissue culture transformations of the root and leaf. The plants were then grown and analyzed using PCR and verified by sequencing for successful gene knockout. Plants demonstrating effective disruption of TPS genes are a step towards understanding the environmental and ecological functions of terpenes in legume plants. The next step would be to subject the mutant plants to herbivore stress and study the response in the terpene-deficient mutants. By understanding the function of terpenes, a door is opened to new opportunities in agricultural applications of this knowledge, such as resistant crops. 

Keywords

CRISPR Mutants , Agricultural applications , Gene editing, Organic compounds , Biochemistry

Research Area

Biological Organisms

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