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International audience; Conditioned both by environment, genotype and the interaction of the genotype itself with the environment, some phenotypic plasticity lies between individuals. This phenomenon allows an adaptive potential of individuals, and may depend on various mechanisms as overdominance, epistasis, pleiotropy, gene linkage and epigenetics. Depending on studied characters, some genotypes or cultivars may be more and less plastic and, in some cases, more and less stable. Considered as powerful anti-oxidants, carotenoids have large applications in medical fields as cancer preventive compounds and precursors of vitamin A and there is an interest for guaranteed carotenoid content in crops. Even if carotenoid accumulation seems to depend on a high genetic component for climacteric fruits as tomato, there is a large effect of G×E interaction overall. Thus, the genetic determinism alone does not explain carotenoid accumulation among most vegetables. Carrot is depicted as a healthy vegetable, thanks to its ability to accumulate carotenoids at high level in roots. However a large variation is observed. In spite of a well-known carotenoid biosynthesis pathway and recent progress on the genetic control, carotenoid accumulation in an underground reserve tissue remains unclear. Regulation can occur at several scales and can result from environmental influences. This article deals with the implication of genotype by environment interaction on carotenoid accumulation and the underlying mechanisms.

International audience; Renewed consumer demand motivates the nutritional and sensory quality improvement of fruits and vegetables. Specialized metabolites being largely involved in nutritional and sensory quality of carrot, a better knowledge of their phenotypic variability is required. A metabolomic approach was used to evaluate phenotypic plasticity level of carrot commercial varieties, over three years and a wide range of cropping environments spread over several geographical areas in France. Seven groups of metabolites have been quantified by HPLC or GC methods: sugars, carotenoids, terpenes, phenolic compounds, phenylpropanoids and polyacetylenes. A large variation in root metabolic profiles was observed, in relation with environment, variety and variety by environment interaction effects in decreasing order of importance. Our results show a clear diversity structuration based on metabolite content. Polyacetylenes, beta-pinene and alpha-carotene were identified mostly as relatively stable varietal markers, exhibiting static stability. Nevertheless, environment effect was substantial for a large part of carrot metabolic profile and various levels of phenotypic plasticity were observed depending on metabolites and varieties. A strong difference of environmental sensitivity between varieties was observed for several compounds, particularly myristicin, 6MM and D-germacrene, known to be involved in responses to biotic and abiotic stress. This work provides useful information about plasticity in the perspective of carrot breeding and production. A balance between constitutive content and environmental sensitivity for key metabolites should be reached for quality improvement in carrot and other vegetables.

National audience; Carotenoids are a large class of plant metabolites with a function of either essential nutrients or health promoting compounds for humans. Carrot root is a well-known and significant source of dietary carotenoids, mainly: alpha- and beta-carotene, lutein and lycopene. These pigments are the main carotenoids separated and quantified routinely by HPLC analysis. However, little is known about minor carotenoids, carotenoid esters and the carotenoids present in leaves despite their potential interest in metabolic and physiological studies. Previous works used C-18 columns but these stationary phases provide a poor resolution of structurally similar compounds and geometrical isomers. In recent years, C-30 columns have been developed and successfully applied at the separation of carotenoids from various plant materials, the number of resolved carotenoids being significantly improved. Based on literature procedures, we have developed a HPLC-DAD method with a C-30 column, adapted to the quantification of carotenoid compounds from carrot roots and leaves. A simple and rapid extraction method was optimized for both these types of samples on a panel of 5 genotypes displaying distinct root colours (different carotenoid composition and contents). Carotenoids from roots were separated in 23 minutes while carotenoids and chlorophylls from leaves were separated in 42 minutes. Compounds were identified according to their retention time and UV-visible spectrum in comparison with authentic standards (analysed individually and in combination, in the same conditions), or with data from literature, when standards were unavailable. Results showed that carrot root exhibited a simple profile with only 1 to 3 main carotenoids whereas a more complex composition was noticed in leaves, containing both identified and unidentified carotenoids and chlorophylls. Moreover, the composition was quite conservative for leaves but depended on the genotype for roots.

International audience; Carotenoids are essential for photosynthesis, and they are the ultimate source of all dietary vitamin A. They account for the striking diversity of orange, yellow, and red carrot storage root color, and this likely contributes to the fact that carotenoids are the most extensively studied class of compounds in carrot, where their biosynthesis and accumulation have been evaluated across diverse genetic backgrounds and environments. Many genes in the 2-C-methyl-D-erythritol-4-phosphate pathway (MEP) and carotenoid biosynthetic pathways have been identified and characterized in carrot, and genes in those pathways are expressed in carrot roots of all colors, including white carrots which contain at most trace amounts of carotenoids. The active functioning of genes in the carotenoid pathway in carrot roots of all colors should be expected since pathway products serve as precursors for hormones important in plant development. 1-Deoxy-d-xylulose-5-phosphate synthase (DXS) in the MEP pathway and the phytoene synthase and lycopene β-cyclase (PSY, LCYB) genes in the carotenoid pathway provide some level of overall regulation or modulation of these respective pathways, and these genes are incrementally upregulated in carrots with higher carotenoid content but variation in their expression does not account for the diverse content and composition of carotenoids in different colors of carrots. In contrast, genetic polymorphism in the Y and Y2 genes accounts for much for the variation in carotenoids accumulated in white, yellow, and orange carrots, and with the sequencing of the carrot genome, the genetic basis for these genes is becoming revealed. A candidate for the Y gene, DCAR_032551, is not a member of either the MEP or carotenoid biosynthesis pathway but rather a regulator of photosystem development and carotenoid storage. A clear candidate for the Y2 gene has not been identified, but no carotenoid biosynthetic gene was found in the genomic region defined by fine mapping of Y2. The Or gene, which regulates chromoplast development in other crops, was also recently associated with the presence of carotenoids in carrot. The discovery of genes outside the carotenoid biosynthetic pathway that contributes to carotenoid colors of carrots is but one exciting consequence of sequencing the carrot genome.

Association mapping becomes one of the major genetic methods used to understand genetic control of complex traits. This method allows the use of natural populations with high resolution thanks to ancestral recombinations. Resolution is given by linkage disequilibrium (LD) extend. But false positives can be detected when phenotypic traits are correlated with underlying population stratification at non-causal loci. Knowledge of population structure is then a preliminary step for association mapping studies. Here the population structure was investigated in cultivated carrot and the LD extend in the carotenoid isomerase gene, a key gene in the carotenoid biosynthesis pathway. An unexpected high LD for outcrossing species was found in this gene. Population stratification analysis confirmed the differentiation of the carrot germplasm in two clusters. The first one comprised European and American accessions, the second one Asian accessions. These are preliminary steps to perform association mapping studies to understand root carotenoid content in cultivated carrot.

UMR BFP - Equipe Metabolisme; International audience; Carotenoids are important secondary metabolites involved in plant photosynthesis, vegetable nutritional quality but also in metabolic prevention in human health. Carrot represents one of the most important carotenoid intakes and is a very interesting model to study carotenoid metabolism. Till now, the knowledge about the impact of stress on carotenoid accumulation is limited. The purpose of this work was to investigate the impact of abiotic and biotic stresses applied separately or in combination on carotenoid accumulation in carrot leaves and roots. Results showed that combined stress decreased dramatically carotenoid content in both organs. In roots, the decrease in carotenoid content could be mostly associated in carrot response to A. dauci infection. Moreover, carotenoid and sugar contents were highly correlated, which suggests that stored metabolites are directly or indirectly involved in plant response to pathogen infection. In leaves, in contrast to results observed in roots, stresses impact carotenoid content depending on the genotype and the year. Moreover, carotenoid content variations were correlated to chlorophyll contents suggesting that a common mechanism of regulation for photosynthetic biosynthesis pigment exists.

Communication (Communication avec actes dans un congrès); International audience; Carotenoids are important plant metabolites considered as beneficial for human health. Provided by fruits and vegetables, they are supposed to be implicated in preventing several chronic diseases. In plants, carotenoids play multiple roles as antioxidant molecules. In light-exposed organs, they also act as photosynthesis accessory pigments and pollinator or disseminator attractors. However, in underground organs, like tuber or root, their role is not clearly understood.Carrot is one of the most important sources of dietary carotenoids due to high content in root. Moreover, the large genetic diversity of this species makes it a suitable model to study carotenogenesis. This diversity results in a differential accumulation of carotenoids: mostly β-carotene, α-carotene, lycopene and lutein, leading to various root colors. Although the carotenoid biosynthetic pathway is well-characterized, basic knowledge of carotenogenesis regulation in roots is still lacking. It is generally considered that carotenoid biosynthesis may be controlled at distinct levels: allelic polymorphism, transcriptional regulation, chromoplast differentiation and root development. Previous works have been conducted to elucidate how those different regulation scales impact carotenoid accumulation in carrots. For instance, single nucleotide polymorphisms detected in carotenoid biosynthesis genes like PSY are correlated with root color. However, little is known about the other types of regulation in carrot. In relation with these intracellular mechanisms, external factors may affect carotenoid accumulation as demonstrated for other plant secondary metabolites.This presentation is an overview of the current knowledge about the regulation of carotenoid accumulation in carrot root and the remaining questions to be addressed.

The preservation of our genetic resources and production of high-quality seeds depends on their ability to remain viable and vigorous during storage. In a quantitative trait locus analysis on seed longevity in Medicago truncatula, we identified the bZIP transcription factor ABSCISIC ACID INSENSITIVE5 (ABI5). Characterization of Mt-abi5 insertion mutant seeds revealed that both the acquisition of longevity and dormancy were severely impaired. Using transcriptomes of developing Mt-abi5 seeds, we created a gene coexpression network and revealed ABI5 as a regulator of gene modules with functions related to raffinose family oligosaccharide (RFO) metabolism, late embryogenesis abundant (LEA) proteins, and photosynthesis-associated nuclear genes (PhANGs). Lower RFO contents in Mt-abi5 seeds were linked to the regulation of SEED IMBIBITION PROTEIN1. Proteomic analysis confirmed that a set of LEA polypeptides was reduced in mature Mt-abi5 seeds, whereas the absence of repression of PhANG in mature Mt-abi5 seeds was accompanied by chlorophyll and carotenoid retention. This resulted in a stress response in Mt-abi5 seeds, evident from an increase in alpha-tocopherol and upregulation of genes related to programmed cell death and protein folding. Characterization of abi5 mutants in a second legume species, pea (Pisum sativum), confirmed a role for ABI5 in the regulation of longevity, seed degreening, and RFO accumulation, identifying ABI5 as a prominent regulator of late seed maturation in legumes.

1432-2048;Perrin, Florent;Hartmann, Laura;Dubois-Laurent, Cecile;Welsch, Ralf;Huet, Sebastien;Hamama, Latifa;Briard, Mathilde;Peltier, Didier;Gagne, Severine;Geoffriau, Emmanuel;Journal Article;Germany;Planta. 2017 Apr;245(4):737-747. doi: 10.1007/s00425-016-2637-9. Epub 2016 Dec 20.; International audience; Main conclusion Variations in gene expression can partially explain the difference of carotenoid accumulation in secondary phloem and xylem of fleshy carrot roots. The carrot root is well divided into two different tissues separated by vascular cambium: the secondary phloem and xylem. The equilibrium between these two tissues represents an important issue for carrot quality, but the knowledge about the respective carotenoid accumulation is sparse. The aim of this work was (i) to investigate if variation in carotenoid biosynthesis gene expression could explain differences in carotenoid content in phloem and xylem tissues and (ii) to investigate if this regulation is differentially modulated in the respective tissues by water-restricted growing conditions. In this work, five carrot genotypes contrasting by their root color were studied in control and water-restricted conditions. Carotenoid content and the relative expression of 13 genes along the carotenoid biosynthesis pathway were measured in the respective tissues. Results showed that in orange genotypes and the purple one, carotenoid content was higher in phloem compared to xylem. For the red one, no differences were observed. Moreover, in control condition, variations in gene expression explained the different carotenoid accumulations in both tissues, while in water-restricted condition, no clear association between gene expression pattern and variations in carotenoid content could be detected except in orange-rooted genotypes. This work shows that the structural aspect of carrot root is more important for carotenoid accumulation in relation with gene expression levels than the consequences of expression changes upon water restriction.

International audience; Carotenoids are important secondary metabolites involved in plant growth and nutritional quality of vegetable crops. These pigments are highly accumulated in carrot root but the knowledge about the environmental factors on their accumulation is limited. The purpose of this work was to investigate the impact of environmental variations on carotenoid accumulation in carrot leaves and roots. In this work, carrots were grown during two contrasted periods to maximize bioclimatic differences. In leaves, carotenoid and chlorophyll contents were lower in the less favorable growing condition while relative contents were well conserved for all genotypes suggesting a common regulatory mechanism. The down-regulation of all genes under environmental constraints demonstrates that carotenoid accumulation is regulated at transcriptional level. In roots, the decrease in α-carotene and lutein contents was accompanied by an increase of β-carotene relative content. At transcriptional level, LCYB and ZEP expression increased while LCYE expression decreased in the less favorable condition, suggesting that carotenoid biosynthesis is switched toward the β-branch.

Article de revue (Article scientifique dans une revue à comité de lecture); International audience; Accumulated in large amounts in carrot, carotenoids are an important product qualityattribute and therefore a major breeding trait. However, the knowledge of carotenoid accumulationgenetic control in this root vegetable is still limited. In order to identify the geneticvariants linked to this character, we performed an association mapping study with a candidategene approach.We developed an original unstructured population with a broad geneticbasis to avoid the pitfall of false positive detection due to population stratification. Wegenotyped 109 SNPs located in 17 candidate genes – mostly carotenoid biosynthesisgenes – on 380 individuals, and tested the association with carotenoid contents and colorcomponents. Total carotenoids and β-carotene contents were significantly associated withgenes zeaxanthin epoxydase (ZEP), phytoene desaturase (PDS) and carotenoid isomerase(CRTISO) while α-carotene was associated with CRTISO and plastid terminal oxidase(PTOX) genes. Color components were associated most significantly with ZEP. Our resultssuggest the involvement of the couple PDS/PTOX and ZEP in carotenoid accumulation, asthe result of the metabolic and catabolic activities respectively. This study brings new insightsin the understanding of the carotenoid pathway in non-photosynthetic organs.

Direct spectrophotometric determination of quercetin content in onions (Allium cepa L.) was investigated as a possible alternative to high-performance liquid chromatography (HPLC) analysis. Quercetin content in five onion varieties was monitored at 362 nm and quantified using simple spectrophotometric and HPLC methods. HPLC revealed that 3,4'-Qdg and 4'-Qmg comprised up to 93% of total flavonol content detected in the studied varieties. These major quercetin conjugates combined (3,4'-Qdg + 4'-Qmg) and total flavonol conjugates quantified by HPLC correlated closely with spectrophotometer values. Correlation coefficients were 0.96 (P < 0.0001) for 3,4'-Qdg + 4'-Qmg and 0.97 (P < 0.0001) for total flavonol conjugates in onion. Simple spectrophotometric procedure proved to be a valid, efficient, and cost-effective method for the quantification of total quercetin in onion. Chemical names used: quercetin-3,4'-O-diglucoside (3,4'-Qdg); quercetin-4'-O-glucoside (4'-Qmg).

International audience; The typically intense carotenoid accumulation in cultivated orange-rooted carrots (Daucus carota) is determined by a high protein abundance of the rate-limiting enzyme for carotenoid biosynthesis, phytoene synthase (PSY), as compared with white-rooted cultivars. However, in contrast to other carotenoid accumulating systems, orange carrots are characterized by unusually high levels of α-carotene in addition to β-carotene. We found similarly increased α-carotene levels in leaves of orange carrots compared with white-rooted cultivars. This has also been observed in the Arabidopsis thaliana lut5 mutant carrying a defective carotene hydroxylase CYP97A3 gene. In fact, overexpression of CYP97A3 in orange carrots restored leaf carotenoid patterns almost to those found in white-rooted cultivars and strongly reduced α-carotene levels in the roots. Unexpectedly, this was accompanied by a 30 to 50% reduction in total root carotenoids and correlated with reduced PSY protein levels while PSY expression was unchanged. This suggests a negative feedback emerging from carotenoid metabolites determining PSY protein levels and, thus, total carotenoid flux. Furthermore, we identified a deficient CYP97A3 allele containing a frame-shift insertion in orange carrots. Association mapping analysis using a large carrot population revealed a significant association of this polymorphism with both α-carotene content and the α-/β-carotene ratio and explained a large proportion of the observed variation in carrots.

Communication (Communication avec actes dans un congrès); International audience; Orange carrots are the example par excellence for extraordinary high and stable accumulation of carotenoids. In contrast to other carotenoid-accumulating systems, knowledge on the molecular mechanisms involved is rather sparse. The first enzyme of the carotenoid pathway, phytoene synthase (PSY), frequently determines carotenoid amounts in many other tissues, and we thus investigated its contribution to carotenoid accumulation in carrots. Overexpression of PSY in white-rooted carrots resulted in the accumulation of b-carotene as crystals. This confirmed that increased phytoene synthesis is sufficient to achieve b-carotene concentrations high enough to result in the formation of crystals. In agreement with this, PSY protein amounts were high in orange but almost absent in white-rooted carrot cultivars. However, this contrasted with only slightly different PSY expression levels comparing white and orange carrots and suggests additional mechanisms acting beyond transcription. In fact, we found a negative feedback regulation emerging from xanthophyll-derived metabolites which modulate PSY protein levels. This was shown by the overexpression of the carotene hydroxylase CYP97A3 from Arabidopsis in orange carrots. These lines showed strongly reduced a-carotene levels through its enhanced CYP97A3-catalyzed hydroxylation, but also lower PSY protein levels which accordingly resulted in lower total carotenoid amounts. In agreement with these findings, we identified an 8 nt frame shift insertion in a carrot CYP97A3 hydroxylase allele encoding a truncated and thus non-functional enzyme. The corresponding polymorphism was significantly associated with high a-carotene levels and high a-/b-carotene ratios determined in a large association mapping analysis and explained a large proportion of the variation in carrots. Our results emphasize the rate-limiting role for PSY in carotenoid biosynthesis and show that previously unknown regulatory mechanisms determine carotenoid accumulation in orange carrots.

Because of potential benefits on human health, the content of quercetin, the major flavonol found in onion (Allium cepa), could become a selection trait in breeding programs. Total flavonol concentration in onion was examined by spectrophotometric analysis at 374 nm in three long-day hybrid cultivars grown at three locations (Parma, Idaho; Grand Rapids, Mich; Elba, N.Y.), and in three shortday hybrid cultivars grown at one location in Georgia in three different fields. Mean total flavonol concentrations for long-day hybrids ranged from 176 to 232 mg·kg-1 (ppm) fresh weight and 110 to 173 mg·kg-1 fresh weight among short-day cultivars. No significant effect of location (state or field) was detected (P > 0.05). A significant (P > 0.05) cultivar by field interaction was detected in the short-day experiment, with the hybrid `Sweet Vidalia' showing significant differences among fields. Overall, our results suggest that quercetin content in onion, as expressed by the total flavonol content, does not vary depending on the growing origin, and therefore could be evaluated effectively in breeding or commercial material.

Carotenogenesis has been extensively studied in fruits and flower petals. Transcriptional regulation is thought to be the major factor in carotenoid accumulation in these organs. However, little is known about regulation in root organs. The root carotenoid content of carrot germplasm varies widely. The present study was conducted to investigate transcriptional regulation of carotenoid biosynthesis genes in relation to carotenoid accumulation during early carrot root development and up to 3 months after sowing. HPLC carotenoid content analysis and quantitative RT-PCR were compared to quantify the expression of eight genes encoding carotenoid biosynthesis enzymes during the development of white, yellow, orange, and red carrot roots. The genes chosen encode phytoene synthase (PSY1 and PSY2), phytoene desaturase (PDS), zeta-carotene desaturase (ZDS1 and ZDS2), lycopene epsilon-cyclase (LCYE), lycopene beta-cyclase (LCYB1), and zeaxanthin epoxidase (ZEP). All eight genes were expressed in the white cultivar even though it did not contain carotenoids. By contrast with fruit maturation, the expression of carotenogenic genes began during the early stages of development and then progressively increased for most of these genes during root development as the total carotenoid level increased in coloured carrots. The high expression of genes encoding LCYE and ZDS noted in yellow and red cultivars, respectively, might be consistent with the accumulation of lutein and lycopene, respectively. The results showed that the accumulation of total carotenoids during development and the accumulation of major carotenoids in the red and yellow cultivars might partially be explained by the transcriptional level of genes directing the carotenoid biosynthesis pathway.

Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699; International audience; Haploidization in onion (Allium cepa L.) using immature flower buds simulates zygotic embryogenesis with no fecundation. In order to know the involvement of polyamines (PAs) in this process, we determined the concentration of endogenous PAs in flower buds and experimented the addition of various combinations of PA molecules in the medium. At the inoculation stage, high levels of free and conjugated spermidine and low putrescine + hydroxyputrescine/spermidine + spermine ratio characterized the highest responsive varieties. During in vitro culture, high levels of putrescine and its derivatives characterized the lowest responsive varieties, whereas high levels of spermidine and spermine characterized responsive varieties. The putrescine + hydroxyputrescine + homospermidine/spermidine + spermine ratio remained low in responsive varieties. The addition of spermidine or spermine (2 x 10(-3) M) to the culture medium improved significantly the embryo production. Our results suggest that the arginine decarboxylase pathway is involved in PA biosynthesis during the in vitro culture of flower buds. Our study showed that specific ratios of PAs are required for successful gynogenesis in onion

Ploidy stability in gynogenic clones of long-day onion is assessed during micropropagation. In order to induce chromosome doubling in haploids, explants and plantlets of three clones were treated in vitro. Colchicine (0.625–12.5 mM) and oryzalin (10–200 μM) were tested for 24 h in combination with culture conditions (4°C in the dark or 24°C in the 16 h day photoperiod). Diploidization performed well (64%) during micropropagation on explants treated with 0.625 mM colchicine at 24°C day. Among treatments on individual plantlets, best results were obtained either with 2.5 mM colchicine (up to 65.7% of diploids) or 50 μM oryzalin (up to 57.1% of diploids). Both chemicals induced mixoploids and affected plant regeneration, but a better plant quality was obtained with oryzalin. Chromosome doubling in explants during the first cycle of micropropagation after gynogenesis appears to be a valuable and time-saving procedure to provide numerous doubled-haploid plants.

International audience; BACKGROUND: Carrot is a vegetable cultivated worldwide for the consumption of its root. Historical data indicate that root colour has been differentially selected over time and according to geographical areas. Root pigmentation depends on the relative proportion of different carotenoids for the white, yellow, orange and red types but only internally for the purple one. The genetic control for root carotenoid content might be partially associated with carotenoid biosynthetic genes. Carotenoid isomerase (CRTISO) has emerged as a regulatory step in the carotenoid biosynthesis pathway and could be a good candidate to show how a metabolic pathway gene reflects a species genetic history. METHODOLOGY/PRINCIPAL FINDINGS: In this study, the nucleotide polymorphism and the linkage disequilibrium among the complete CRTISO sequence, and the deviation from neutral expectation were analysed by considering population subdivision revealed with 17 microsatellite markers. A sample of 39 accessions, which represented different geographical origins and root colours, was used. Cultivated carrot was divided into two genetic groups: one from Middle East and Asia (Eastern group), and another one mainly from Europe (Western group). The Western and Eastern genetic groups were suggested to be differentially affected by selection: a signature of balancing selection was detected within the first group whereas the second one showed no selection. A focus on orange-rooted carrots revealed that cultivars cultivated in Asia were mainly assigned to the Western group but showed CRTISO haplotypes common to Eastern carrots. CONCLUSION: The carotenoid pathway CRTISO gene data proved to be complementary to neutral markers in order to bring critical insight in the cultivated carrot history. We confirmed the occurrence of two migration events since domestication. Our results showed a European background in material from Japan and Central Asia. While confirming the introduction of European carrots in Japanese resources, the history of Central Asia material remains unclear.

The flavonol quercetin has been reported as having many health benefits, including a reduction in the risk of developing cardiovascular disease, stroke, and some types of cancer. The overall content of quercetin in onion was examined in four yellow varieties (`Rio Rita', `RNX 10968', `Predator', and `Tamara') and one red variety purchased at a local grocery store. Each bulb was quartered, with one quarter saved as a control and the other three quarters subjected to three cooking treatments that simulated common domestic processing methods of preparing onion. The treatments included sautéing in sunflower oil for 5 minutes at 93 °C, baking for 15 min. at 176 °C, and boiling for 5 minutes in distilled water. Samples were frozen in liquid nitrogen, ground to a fine powder, blended with 80% EtOH, and filtered for quercetin extraction. The filtrate was then analyzed with a spectrophotometer (uv 374 nm). Quercetin concentrations were obtained in mg quercetin/kg fresh weight of tissue by regressing spectrophotometer readings onto a standard curve. Significant differences between varieties were found when examining fresh samples alone with the red variety containing the highest content of quercetin. Results of cooking showed that sautéing produced an overall 27% gain in quercetin concentration (significantly higher than the fresh control), baking produced an overall 4% gain in concentration (insignificant from the fresh control), while boiling produced an overall 18% loss in quercetin concentration (significantly lower than the fresh control).