Summary Cellular oxidative stress is a risk factor for several human disease including diabetes, cardiovascular diseases (CVD) and cancer. Natural and food products have a great potential to protect cells under oxidative stress conditions. Tef (Eragrostis tef) is gluten-free grain that is rich in mineral nutrients, essential amino acids, and water-soluble vitamins and several bioactive phytochemicals including polyphenols, flavonoids and polyunsaturated fatty acids (PUFAs). Tef has been recognized by the U.S. National Research Council as a crop with excellent nutritional potential and suitability to boost food security. However, the bioactivity of tef phytochemicals have never been studied using physiologically relevant models. In this study, we propose to identify bioactive compounds in tef grains and test their ability to combat oxidative stress, thus preventing the pathogenic conditions associated with reactive oxygen species (ROS)-induced damages. This initiative is based on our recent finding indicating that: 1) tef seed extracts increase glutathione (GSH) levels in THP-1 monocytes, 2) an increase in GSH level is more prominent in brown than ivory tef seeds, suggesting the existence of genotypic variability in the content of bioactive compound, 3) Tef seed extracts increase the expression of the master regulator of antioxidant pathway nuclear factor erythroid 2-related factor 2 (Nrf2) and its targets including Heme Oxygenase 1 (HO-1), NAD(P)H quinone oxidoreductase1 (NQO1), glutathione reductase (GR) and the γ-glutamate cysteine ligase catalytic (GCLC) and regulatory (GCLM) subunit, which play a key role in GSH biosynthesis, 4) A bioactive fraction of tef extract contains a derivative of linoleic acid (LA) 9-Hydroxyoctadecadienoic acid (9-HODE), which is a marker for membrane lipid peroxidation. These findings prompted us to hypothesize that tef has bioactive compounds which have the potential to boost the antioxidant pathway under oxidative stress conditions likely via Nrf2 signaling. Our aims are: 1) Identification and functional characterization of bioactive phytochemicals from tef seeds, 2) Studying whether tef bioactive compounds induce oxidative stress-responsive genes, and protect cells from ROS, 3) Understanding the genetic mechanism regulating antioxidant properties of tef. This study will help in developing alternative medicines for oxidative stress-induced diseases. It will also generate new information that can be used to improve tef or transferred to major cereals to add value to these crops. This funding will boost the capacity of the Osena laboratory to explore underutilized crops such as millets for desirable traits including health benefits. The Osena laboratory and the environment at UNC Greensboro are ideal for this research, and training students from underrepresented minority group.

Projective narrative The rising super crop tef has a great potential as nutritious and healthy grains, but there has been limited knowledge available on this crop due to lack of investment in its research and limited genome information. This proposal aims to identify bioactive chemicals from tef grains and study their role in boosting the antioxidant pathway towards mitigating ROS-induced inflammatory diseases such as CVD, type II diabetes and cancer. We also aim to identify the molecular genetic mechanisms regulating variation in antioxidant activity in tef grains that can be used to improve tef or introduced into other major crops such as corn via molecular breeding.