Identifying the Biosynthetic Pathways for Pharmaceutical alkaloids in Mitragyna species

Swathi Nadakuduti, Principal Investigator on the USDA-NIFA grant, in her greenhouse facility along with her PhD candidate, Larissa Laforest (in orange) from Plant Molecular and Cell Biology Program.

A multidisciplinary team of researchers from UF/IFAS, College of Pharmacy and UBC, Canada will work on pathway discovery of pharmaceutical alkaloids in kratom. The leaves of Mitragyna speciosa (kratom), produce more than 50 monoterpene indole alkaloids (MIAs) and spirooxindole alkaloids associated with varied pharmaceutical uses. The MIA mitragynine has shown promise as a potential treatment for pain, opioid use disorder, and opioid withdrawal without any demonstrated addiction potential; and the spirooxindole mitraphylline shows anti-tumor activity. The existing knowledge gap in the biosynthesis of bioactive M. speciosa alkaloids prevents systematic advances in engineered production of these high value compounds, creating a bottleneck for pharmaceutical research and production. A multi-faceted approach incorporating genomics, transcriptomics, and metabolomics will be jointly applied to enable pathway discovery of diverse bioactive alkaloids with the goal of supporting synthetic biology efforts to produce these alkaloids in appreciable quantities for clinical study.

Exploring Florida Native plants for metabolites of pharmaceutical value

We are interested in exploring medicinally relevant species of Florida natives for plant-specialized metabolites to identify bioactive compounds, their spatio-temporal patterns of accumulation and decipher metabolic pathways.

Scutellaria, a Florida native ornamental with potential anti-cancer properties

Scutellaria spp has tremendous potential for the ornamental and medicinal plant industry in Florida. The root extract of S. baicalensis has been widely used for medicine in Asia for over 2,000 years, and it has a variety of bioactivities, e.g., anti-inflammatory, anti-cancer and neuro-psychologic properties due to synthesis of unique flavonoids and indolic compounds. Commercially, it has a market for ornamental and medicinal applications including supplement capsules, dried plants for tea, powdered or liquid extracts. Florida native species, S. integrifolia and S. arenicola also synthesize these active medicinal compounds and in order to develop Scutellaria crop as a new enterprise crop, we need to identify germplasm and best production practices, while evaluating the metabolite profiles under controlled environments to maximize the synthesis of active pharmaceuticals. This project is a part of a new UF team “MICE (Medicinals In Controlled Environments)” across UF IFAS and College of Pharmacy

As a part of “Support for Emerging Enterprise Development Integration Teams” (SEEDIT), we will evaluate Florida native Scutellaria species in FL landscape locations for ornamental & medicinal purposes, manipulate controlled environment in order to maximize pharmaceutically active metabolites, quantify price and quality of existing Scutellaria products and develop enterprise budget and communicate project outcomes with producers, processors and entrepreneurs.

Saw palmetto, a Florida native palm used as a largest herbal supplement globally

Saw palmetto, Serenoa repens is a Florida native palm and the berries are wild harvested throughout the state of FL. Saw palmetto is the third most utilized dietary supplement in the US for benign prostatic hyperplasia and lower urinary tract symptoms. S. repens grows in a variety of habitats, including sandy coastal lands, pine flatwoods, and under story areas in hardwood hammocks in FL. We are interested in exploring the genetic diversity and correlate with chemical diversity (lipido-sterolic composition) to understand the pharmaceutical potential of berries across the FL geography.

Dissecting molecular mechanisms to enhance fruit photosynthesis and nutritional quality of tomato.

Swathi Nadakuduti, Principal Investigator on the USDA-NIFA grant is working on improving the fruit nutritional quality using tomato as a model system. Although leaves are the primary photosynthetic organs, up to 20% of the carbon in tomato fruit is derived from fruit photosynthesis itself, impacting yield, nutrient content, flavor, and overall fruit quality. PI's research identified TKN4 in tomato, a Class I KNOTTED-like homeobox (KNOX) gene that also influences chloroplast development and chlorophyll accumulation in fruit. Tomato uniform ripening (u) and uniform grey green (ug) mutants showing differential chlorophyll accumulation will be used in identifying the KNOX regulatory network that has the long-run potential to favorably engineer photosynthesis contributing to yield and quality of food crops.

Genome-wide evaluation of off-targets from gene editing reagents in seed vs. vegetatively propagated crop species

Gene-editing by CRISPR systems provide an unprecedented advancement in genome engineering due to precise DNA manipulation. Genome-editing is being widely applied in plants and has revolutionized crop improvement by offering the ability to generate transgene-free plants. Tremendous progress has been made recently to increase the specificity of CRISPR reagents by structure-guided protein engineering to develop variants of Cas9 such as hyper-accurate Cas9 (Hypa Cas9). As alternative CRISPR-Cas12a is also emerging as an effective reagent for gene-editing in plants. Base-editing technology based on CRISPR/Cas9 system generates base substitutions without requiring dsDNA cleavage. There are no studies to date that characterize genome-editing reagents’ specificity in plants. The recently streamlined regulatory processes by the USDA opened up new opportunities for scientists to develop improved crops and useful agricultural products. Therefore, there is a need to be able to effectively assess this emerging technology in terms of genome-wide off-target effects before it is applied to new crop varieties. As a part of USDA – Biotechnology risk assessment Grant project, we will use CRISPR/Cas9, Cas12a, HypaCas9, base editors and TALENs to target economically important candidate genes in vegetatively propagated potato and seed propagated Camelina sativa, an alternative oilseed crop in order to quantify the frequency of off-target variation caused by gene-editing reagents by whole genome sequencing. This study would provide comprehensive risk assessment of gene-editing in crop species.