Abstract Format Guidelines

1) Use ARIAL10 and single line spacing.

2) Type the abstract title in bold font.

3) Start a new line for authors’ names, using superscript numbers to denote affiliations, and underline name of presenting author.

4) Provide addresses for the various institutions of the authors, preceding each address with the appropriate superscripted number.

5) Provide e-mail addresses of the presenter and corresponding authors.

6) Leave one line blank. 7) Type the abstract text (maximum 300 words). Do not include any graphics or tables. Please follow the example below.

Example:

Production of bioactive isoflavones in elicited hairy root cultures of pigeon pea

Gaurav Gajurel1,2, Amit R. Sharma2, Salma Abdel-Karim2, Carlos Balmaceda2, Mohammad A. Alam3, and Fabricio Medina-Bolivar1,4. 1Molecular Biosciences Graduate Program, 2Arkansas Bioscience Institute, 3Department of Chemistry and Physics, 4Department of Biological Sciences, Arkansas State University, Jonesboro, AR 72401. E-mail: gaurav.gajurel@smail.astate.edu; fmedinabolivar@astate.edu

Isoflavones, a class of flavonoids, have anticancer, antioxidant, anti-inflammatory, antibacterial, and pro-apoptotic properties. Extracting and purifying these bioactive phenolics from natural sources is challenging and time-consuming. This project aimed to develop a hairy root culture system for pigeon pea using Agrobacterium rhizogenes-mediated transformation as a sustainable production platform for isoflavones. Twelve-day-old hairy root cultures were co-treated with methyl jasmonate, methyl-β-cyclodextrin, hydrogen peroxide, and magnesium chloride for 192 hours to examine their effects on isoflavone accumulation in the culture medium and root tissues. Two isoflavones, genistein and its prenylated analog isowighteone, were identified and purified from the elicited culture medium extract using column chromatography, semi-preparative high-performance liquid chromatography, tandem mass spectrometry, and 1D & 2D nuclear magnetic resonance spectroscopy. Morphological differences between normal and elicited hairy roots were studied using scanning electron microscopy. Elicited hairy root tips showed non-uniform cell shapes and surface ruptures. The antioxidant activity of these isoflavones and isoflavone-rich extracts was measured using the DPPH assay, with the extract showing higher antioxidant activity than individual compounds at equimolar concentrations. The anticancer activities of genistein and isowighteone at 25 micromolar were tested in MDA-MB-231 triple-negative and MCF-7 estrogen receptor-positive breast cancer cells, whereas the anti-inflammatory activity was assessed in RAW 264.7 macrophages. Isowighteone demonstrated higher cytotoxicity against MDA-MB-231 and MCF-7 cells up to 72 hours and greater anti-inflammatory activity at 24 hours compared to genistein. The antibacterial activity of genistein and isowighteone was tested against various gram-positive and gram-negative bacteria, with isowighteone showing higher activity against gram-positive bacteria. In summary, pigeon pea hairy roots provide a sustainable platform for producing bioactive isoflavones, and the prenylated isoflavone isowighteone showed higher biological activities than its non-prenylated analog genistein.