Performing Department
Food & Animal Sciences
Non Technical Summary
PROJECT SUMMARYInterest in the sensory aspects and acceptance of edible plants, in particular the daylily cultivar (Hemerocallis, spp) continues to grow due to consumer interest in the food, culinary, health, functional and medicinal properties of these plants.There are over 52,000 known daylily cultivars, all are edible.Daylilies are an underutilized food source. Consumers do not know how to use them for food.The sensorial and functional properties have not been thoroughly researched.The acceptance of edible daylily (Hemerocallis, spp), is dependent on consumer awareness.Additional value-added benefits such as the functional and medicinal properties of such plants will allow opportunity for increased consumer usage resulting in increased revenue for growers and businesses.During the period of this project, the objectives to be carried out will be to grow edible angiosperm (Angiospermae) daylily cultivars (Hemerocallis, spp.) in segregated plots, determine the optimum harvest time for preferred sensory attributes of the selected cultivars, develop a sensory descriptive language for each attribute category, define the functional and value-added applications of such cultivars by incorporation into novel foods as extracts, dried powders, and pastes, investigate medicinal and toxicological properties of edible plants, determine consumer acceptance of new products developed containing the edible plants, and investigate the bioactivities for safety and potential cytotoxicity.This research will provide investigation into: the usage of daylilies as a food source, development of a sensory lexicon, acceptance, and value-added applications as food ingredients.This research will provide educational opportunities for students and information for the agricultural and food community.
Animal Health Component
0%
Research Effort Categories
Basic
(N/A)
Applied
50%
Developmental
50%
Goals / Objectives
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.
Project Methods
Selection and Growing of Daylily (Hemerocallis, spp.) Cultivars Selections will be made among the more than 35,000 available cultivars for types that will flourish in Alabama. 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. Initial plots will be planted in a complete block design in two locations at the WTARS to represent full sun and partial shade. 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.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). Quantitative Descriptive Analysis of Daylily (Hemerocallis spp.) Cultivars Formal sensory analysis using the quantitative descriptive analysis (QDA) methodology will be performed. At least five panelists will be trained on QDA procedures and will be consistently calibrated and refined using reference standards. 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 scale for references where, 0 = attribute not detected; and 15 = attribute extremely detected from an intensity level. The objective of this portion of the study is to develop the sensory lexicon for the cultivars tested. At least fifteen daylily cultivars will be analyzed using quantitative descriptive analysis (QDA). Daylily flowers will be harvested on the days of QDA panels and held for a minimum time, less than two hours. Teflon® sniff bottles will be utilized for the olfactory portion of the QDA panels. Panelists will develop the terminology that will be used to generate the sensory lexiconal language for the daylily cultivars. Sensory acceptance and product development of beverages containing Hemerocallis powder. Incorporation of Hemerocallis powder into beverages will be evaluated for sensory acceptance. Hemerocallis sepals and tepals will be oven dried then pulverized using a Waring spice grinder to a minimum of 60 mesh sizing. 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.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. A 5 KN load cell and a Warner-Bratzler and a cylindrical punch attachments will be used. Instrumental textural profile parameters obtained by cyclic compression of the samples to 75% of their original thickness. Toughness, which is the ability of the product to withstand repeated stress before fracture, will be measured as the area under the first curve of ITPA.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 solutions to establish equilibrium moisture content (EMC). Hence, EMC vs Aw will be established and the GAP mathematical model will be used to fit the model.Shelf 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 timesDetermination 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. Thus, the shelf-life will be defined as the length of time the flower maintains acceptable appearance for marketability. 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). Reduced (GSH) forms (nmol/mg protein) will be measured using a spectrophotometric assay (Griffith, 1985). The experiment will be repeated three times and samples will be analyzed in triplicates.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).