Sensory Evaluation and Value-added Applications of Edible Daylilies (hemerocallis, Spp)

RESEARCH OBJECTIVESTo grow select cultivars of edible daylilies (Hemerocallis, spp) for the exploration of sensory evaluation, value-added agricultural purposes, food engineering applications and toxicological studies on safety and cytotoxicity. Subtitle: Selection and Growing of Daylily (Hemerocallis, spp.) Cultivars Experimental plots will be established at the Alabama A&M University, Winfred Thomas Agricultural research Station (WTARS), with emphasis on genotypic diversity for continuous and abundant flowering. Selection of cultivars will be assessed on degree of the secondary metabolites anthocyanins that originate from the flavonoid biosynthesis pathway (Hall et al., 2015). The establishment of daylily plots at the WTARS will begin with a cursory analysis using a minimum of 50 cultivars with broad genetic variations. Post-harvest analysis of flowers will be carried out to determine desirable functional chemical profile. Desired types will be selected for further analysis and
parentage for hybridization to create unique cultivars with enhanced desirable characteristics.To develop a sensory lexicon for daylily cultivars, whereby developing complete sets of descriptive attributes for the following categories: appearance, smell, flavor, basic tastes, feeling factors, texture (hand-held), texture (mouth) for select daylily cultivars.Subtitle: Sensory Ranking Procedure of Select Daylily (Hemerocallis, spp): The flowers of at least 15 daylily cultivars will be ranked according to overall preference. The top five ranked cultivars will be chosen for formal sensory studies and value-added applications to foods. The daylily flowers will be hand harvested based on the harvesting method of Pollard (2006).Subtitle: Quantitative Descriptive Analysis of Daylily (Hemerocallis spp.) Cultivars At least five panelists will be trained on QDA procedures and will be consistently calibrated and refined using reference standards. Panelists will be trained for no less than 36
hours. Daylily cultivars will be evaluated descriptively for appearance, aroma, flavor, basic tastes, texture (hand-held), texture (fork), texture (oral), feeling factors, and chew rate. Training will be performed using a 15-point intensity scale for references where, 0 = attribute not detected; and 15 = attribute extremely detected from an intensity level. Panelists will develop the terminology that will be used to generate the sensory lexiconal language for the daylily cultivars.Determination of value-added applications for Hemerocallis by incorporating the daylily components into foods as functional agents, colorants, texturizers, flavorings, or other comprised value-added additions. Subtitle: Sensory acceptance and product development of beverages containing Hemerocallis powder. Incorporation of Hemerocallis powder into beverages will be evaluated for sensory acceptance. Physical analyses will be performed on the beverages, i.e. rheological studies, color studies (Hunter color).
Beverages will be evaluated for sensory acceptance of preferred cultivar.Subtitle: Instrumental Textural profile analysis of the Hemerocallis cultivars to evaluate and establish a correlation between subjective sensory observations to Instrumental Textural Profile Analysis (ITPA) during the storage period of Hemerocallis cultivars Instron Universal Testing Apparatus, (ATS Series model 1601, Warren, MI) will be used to obtain textural profile of the Hemerocallis cultivars samples.Subtitle: To maximize the fresh-life of Hemerocallis cultivars via environmental controlled storage studies to optimize the shelf life and the development of a sorption isotherm. Isotherm Development: A desorption sorption isotherm will be established for each cultivar. Gravimetric methods of establishing a desorption isotherm are based on exposing the samples in a controlled environment using salt solutionsShelf Life Studies: The samples (15 cultivars) will be placed in the Environmental chamber (EC) (ESPEC
Model BTL 433-4 CU. ft, Hudsonville, Michigan). Hemerocallis cultivars will be stored in the EC at 15°C and 90 and 95% Relative Humidity (HR). The samples will be stored in EC for 3, 6, & 9 days. Each experimental unit will be replicated three timesTo determine the optimum month for consumption and storage quality of Hemerocallis cultivars. Subtitle: Determination of the optimum month for consumption of Hemerocallis grown in northern Alabama: Observance will be performed using an observance quality scale. The observance scale will have quality score selections from one to five, with one being "unmarketable" and five being "fresh market acceptable". The daylilies will be harvested at one-hour intervals and the observation scale of quality will be performed on each immediately prior to the formal sensory descriptive analysis will be performed. The flowers will be observed each day following harvest to determine length of acceptable marketability. 5: To determine the
bioactivities (safety and potential cytotoxicity) of daylily cultivars Hemerocallis, spp Subtitle: Determination of the bioactivities (toxicity and cytotoxicity) of daylily cultivars (Hemerocallis spp) grown in northern Alabama: In this objective the biological activities (safety and toxicity) of 15 daylily cultivars will be determined. We will identify varietal differences on bioactivity in two cancer cell lines (Caco-2 and Hep 2G) and non-cancer cell line (Thle-3). We will investigate anticancer properties i.e., proliferation inhibition, apoptosis inducing and anti-inflammatory activities, modulations to antioxidants enzymes that play crucial roles in several chronic diseases such as cancer, diabetes and obesity.Cytotoxicity properties of Daylily cultivarsCytoxicity determination by LDH measurements: LDH release will be assayed using a colorimetric cytotoxicity detection kit (LDH) (Roche Diagnostics, Indianapolis, IN) according to the manufacturer's instructions. Absorbance
will be recorded at 490nm and experiment will be performed in triplicates.Daylily extracts: The antiproliferative activity of lipids will be determined by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay Caco-2, Hep 2G and Thle-3 cells will be seeded at a density of 0.9 × 103 cells/well into a 96-well plate in final volume of 100ul of media. Anti- proliferative properties of the daylily extracts will be determined by reading the absorbance of the reduced intracellular formazan product at 550/630 nm on a synergy HT micro plate reader. Assay will be performed in triplicates.Effect of Daylily extracts on endogenous antioxidants Enzymatic and non-enzymatic assays (in vitro and in vivo experiment): CAT activity will be measured by the reaction of formaldehyde produced from methanol with Purpald to produce a chromophore according to the method of Johansson and Hakan Borg (1988).Anti-inflammatory properties of Daylily extracts.Caco-2, Hep 2G and Thle-3 cells
culture supernatants from the basolateral side will be collected and pooled for analysis of inflammatory markers. Supernatants will be applied to membranes containing cytokine antibody array (Ray Biotech) for 1 h at room temperature, and detection will be performed with horseradish peroxidase-coupled secondary antibodies and chemiluminescence substrate (Ray Biotech). Array spot densities will be evaluated and compared using Quantity One software (Bio-Rad, Hercules, CA). Cell inflammatory markers TNF-α, IL-1β, IL-4, IL-6, IL-10, IL-11, IL-12 and IL-13 will be measured.