Superior Flavor Research Articles

Multi-residue analytical method development and dietary exposure risk assessment of 345 pesticides in mango by LC-Q-TOF/MS

Mango is attracting increasing global attention for its superior flavor and nutritional benefits. However, the use of pesticides has resulted in a threat to its quality, safety, and human health due to their inevitable residue. There is an urgent need for advanced technologies that can efficiently and sensitively analyze a wide range of pesticides. In this study, a high-throughput screening and quantification method for 345 pesticides in mango was established by applying a modified QuEChERS method combined with liquid chromatography-quadrupole-time-of-flight mass spectrometry (LC-Q-TOF/MS). A response surface methodology (RSM) optimized the critical parameters for extracting various pesticides in 1% acetic acid-acetonitrile (v/v) using MgSO4, PSA, C18, and GCB as the adsorbents. The proposed method was successfully validated and applied to the quantitative detection of pesticide residues in 73 mango samples from Hainan. The study found pesticide residues in 95.9% of mango samples, of which 3 samples had residue concentrations in excess of the Chinese maximum residue limit. A total of 24 pesticides were detected, with detection percentages ranging from 1.4% (Thiabendazole) to 41.1% (Pyraclostrobin). Considering the possible health risks related to pesticide residues, risk assessment of human exposure to pesticides via intake of mango was evaluated. The acute and chronic dietary exposure risks related to pesticide residues in mango were accepted. In conclusion, this study provides a powerful platform for monitoring multiple pesticides in mango. First, it can promote the rational use of pesticides, and second, it can help consumers to assess the risk of dietary exposure to pesticide residues.

Effect of Ganoderma lucidum water extract on flavor volatiles and quality characteristics of set-type yogurt

This study investigated the effects of several concentrations of Ganoderma lucidum water extract (GLWE) (0, 0.5, 1.0, and 1.5 %) on set-type yogurt's flavor volatiles as well as the physicochemical, textural, and antioxidant activities of the yogurt during storage. The HS-SPME-GC–MS investigation found that adding GLWE increased the amount of flavor volatiles in yogurt, which improved the quality of flavor volatiles produced by the yogurt. The yogurt's water holding capacity, syneresis, color, and texture all showed optimum values at a 0.5 % GLWE concentration, and its total number of flavor volatiles reached 43. To sum up, the incorporation of GLWE at a 0.5 % concentration promotes the development of superior flavor volatiles in yogurt and enhances its qualitative attributes in contrast to the control group.

Seasonal variation in non-volatile flavor substances of fresh tea leaves (Camellia sinensis) by integrated lipidomics and metabolomics using UHPLC-Q-Exactive mass spectrometry

Harvest season exerts great influence on tea quality. Herein, the variations in non-volatile flavor substances in spring and summer fresh tea leaves of four varieties were comprehensively investigated by integrating UHPLC-Q-Exactive based lipidomics and metabolomics. A total of 327 lipids and 99 metabolites were detected, among which, 221 and 58 molecules were significantly differential. The molecular species of phospholipids, glycolipids and acylglycerolipids showed most prominent and structure-dependent seasonal changes, relating to polar head, unsaturation and total acyl length. Particularly, spring tea contained higher amount in aroma precursors of highly unsaturated glycolipids and phosphatidic acids. The contents of umami-enhancing amino acids and phenolic acids, e.g., theanine, theogallin and gallotannins, were increased in spring. Besides, catechins, theaflavins, theasinensins and flavone/flavonol glycosides showed diverse changes. These phytochemical differences covered key aroma precursors, tastants and colorants, and may confer superior flavor of black tea processed using spring leaves, which was verified by sensory evaluation.

Change in volatile profiles of wheat flour during maturation

The volatile profiles of wheat flour during maturation were examined through headspace solid-phase micro-extraction gas chromatography-mass spectrometry (HS-SPME-GC/MS) combined with electronic nose (E-nose) and electronic tongue (E-tongue) analyses. The wheat flour underwent maturation under three distinct conditions for predetermined durations. While GC/MS coupled with E-tongue exhibited discernment capability among wheat flour samples subjected to varying maturation conditions, E-nose analysis solely relying on principal component analysis failed to achieve discrimination. 83 volatile compounds were identified in wheat flour, with the highest abundance observed in samples matured for 50 d at 25 °C. Notably, trans-2-Nonenal, decanal, and nonanal were the main contributors to the characteristic flavor profile of wheat flour. Integration of HS-SPME-GC/MS with E-tongue indicated superior flavor development and practical viability in wheat flour matured for 50 d at 25 °C. This study furnishes a theoretical groundwork for enhancing the flavor profiles of wheat flour and its derivative products.

Multi-omics analysis unravels chemical roadmap and genetic basis for peach fruit aroma improvement

Selection of fruits with enhanced health benefits and superior flavor is an important aspect of peach breeding. Understanding the genetic interplay between appearance and flavor chemicals remains a major challenge. We identify the most important volatiles contributing to consumer preferences for peach, thus establishing priorities for improving flavor quality. We quantify volatiles of a peach population consisting of 184 accessions and demonstrate major reductions in the important flavor volatiles linalool and Z-3-hexenyl acetate in red-fleshed accessions. We identify 474 functional gene regulatory networks (GRNs), among which GRN05 plays a crucial role in controlling both red flesh and volatile content through the NAM/ATAF1/2/CUC (NAC) transcription factor PpBL. Overexpressing PpBL results in reduced expression of PpNAC1, a positive regulator for Z-3-hexenyl acetate and linalool synthesis. Additionally, we identify haplotypes for three tandem PpAATs that are significantly correlated with reduced gene expression and ester content. We develop genetic resources for improvement of fruit quality.

Honeybee (Apis mellifera L.) pollination enhances the yield and flavor quality of kiwifruit.

Pollination is essential for achieving high yields and enhancing the quality of kiwifruit cultivation, both of which significantly influence growers' interests and consumers' preferences. However, compared to studies on yield, there are fewer studies exploring the impact of pollination methods on the flavor of kiwifruit Actinidia chinensis Planchon. This study examined the effects of bee (Apis mellifera L.) pollination and artificial pollination on the yield and flavor of kiwifruit in the main producing areas of China. Compared with those pollinated artificially, bee-pollinated kiwifruit exhibited a greater fruit set rate, heavier fruit weight, and greater number of seeds. Notably, the number of seeds was positively correlated with fruit weight in bee-pollinated kiwifruit, whereas no such correlation was detected in artificially pollinated fruit. Bee pollination not only enhanced the yield but also improved the flavor of kiwifruit. Specifically, bee-pollinated kiwifruit contained higher levels of sucrose and lower concentrations of glucose and fructose, while the acid content was less affected by pollination methods. Furthermore, significant differences were observed in the volatile organic compound (VOC) levels in kiwifruit subjected to different pollination treatments, with bee-pollinated fruit exhibiting a superior flavor. Our findings provide new insights into the beneficial role of bee pollination in enhancing kiwifruit yield and quality, underscoring the crucial importance of bees in kiwifruit pollination.

Utilizing deep learning via computer vision for agricultural production quality control: jackfruit growth stage identification

Jackfruit (Artocarpus heterophyllus), a tropical fruit renowned for its diverse culinary uses, necessitates identifying the optimal growth stage to ensure superior flavor and texture. This research investigates employing deep learning techniques, particularly convolutional neural networks (CNNs), for accurately detecting jackfruit growth stages. Despite the challenge posed by the nuanced visual differences among fruits at various maturity stages, a meticulously curated dataset of labeled jackfruit images was developed in collaboration with experts, utilizing the BBCH scale. This dataset facilitated training and evaluation. A modified version of the Places 365 GoogLeNet CNN model was proposed for classifying four distinct growth stages of jackfruit, compared with a state-of-the-art CNN model. The trained models demonstrated varying levels of accuracy in classification. Furthermore, the proposed CNN model was trained and tested using original and augmented images, achieving an impressive overall validation accuracy of 90%. These results underscore the efficacy of deep learning in automating the detection of growth stages, offering promising implications for quality control and decision-making in jackfruit production and distribution.

The kinetic study of 2-acetyl-1-pyrroline accumulation in the model system: An insight into enhancing rice flavor through the Maillard reaction

Controlling the Maillard reaction may affect the generation of 2-acetyl-1-pyrroline, the key aroma compound in rice. In this study, the kinetics of 2-acetyl-1-pyrroline accumulation in the glucose/proline model system was comprehensively investigated and extra methylglyoxal or glyoxal was added to enhance 2-acetyl-1-pyrroline concentrations during rice cooking. Using the multi-response kinetic modeling to derive kinetic parameters, the formation of glyoxal, as the reactive intermediate, was rate-determining for the overall generation rate of 2-acetyl-1-pyrroline. Besides, although 2-acetyl-1-pyrroline generation was easier to occur with lower activation energy, much higher depletion rates of 2-acetyl-1-pyrrroline at 120 °C and 140 °C led to maximal 2-acetyl-1-pyrroline accumulation at the lower temperature of 100 °C. Furthermore, the inclusion of 0.05 μmol/kg additional methylglyoxal in cooked rice significantly enhanced 2-acetyl-1-pyrroline generation. The work suggested that the development of rice products with superior flavor quality may be achieved by the slight accumulation of intermediates prior to thermal processing.

A comparative analysis of growth and nutritional quality of the hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) in a simulated pond system and a recirculating aquaculture system

Groupers, as a popular economic fish species, are now more frequently cultured in land-based facilities, however, traditional pond-farming methods are plagued by frequent disease outbreaks, unstable economic benefits, and environmental pollution. To explore a new farming mode for a grouper, an 80-day cultivation experiment was conducted using both a simulated pond system (SPS) and a recirculating aquaculture system (RAS). The research aims to evaluate the growth performance, health (pathogenic bacteria, intestinal microbiota), off-flavor compounds (geosmin, 2-methylisoborneol), and nutritional component (amino acids, fatty acid composition and content) of a hybrid grouper (Pinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) under different aquaculture systems. The results showed that the hybrid grouper in a RAS exhibited better growth performance; the concentration of Vibrio in the fish tissue in the RAS was significantly lower than that in the SPS. Moreover, the content of fresh amino acids in the RAS was significantly higher, and the levels of saturated fatty acids (SFAs) and ω-6 polyunsaturated fatty acids (ω-6PUFAs) were significantly higher in the RAS. This finding indicates the superior flavor and nutritional value of the grouper. These results prove that the RAS is suitable for the widespread cultivation of grouper.

Genotyping of cacao (Theobroma cacao L.) germplasm resources with SNP markers linked to agronomic traits reveals signs of selection

The Amazonian cacao tree or the chocolate tree (Theobroma cacao L.) is cultivated to produce seeds, the valuable raw material for the chocolate industry. However, cacao yield is hampered by diseases and low productivity. Cacao single nucleotide polymorphism (SNP) markers associated with breeding and adaptive traits provide a genetic tool for improving selection and reducing breeding cycles. Here, we genotyped 40 published SNPs associated with disease resistance and 11 SNPs with yield traits in 346 accessions using the MassARRAY® system. These 51 SNPs were identified in different breeding experiments in Costa Rica, Brazil, and Trinidad and Tobago. Four genetic clusters were identified, and two of them were observed in high proportion in managed cacao, characterized by a high proportion of admixed individuals reflecting the man-made hybrids. One cluster overrepresented in managed cacao is associated with the preference to keep superior flavor in cultivated cacao from Criollo ancestry. Introgression of wild material collected in Peru associated with disease resistance to witches’ broom disease (WBD) was also observed. Underutilized genetic resources were observed in managed cacao such as the Guiana cacao, while previous agronomic evaluation has demonstrated its good yield potential. Yield and disease resistance traits (mainly resistance to WBD) show divergence between wild and managed cacao (FST > 0.05) probably reflecting selection during domestication, cultivation, and breeding efforts. Eleven SNPs showed a divergence between wild and managed cacao and can be used to build breeding populations. New collections of wild cacao followed by agronomic evaluations can broaden the genetic base of cultivated cacao, especially for such traits as disease resistance to frosty pod rot (FPR).

Gas chromatography–mass spectrometry‐based electronic nose, glycosidically bound volatile, and alcohol‐acyltransferase activity profiles of Okinawan pineapple

SummaryA recent increase in the demand for pineapple necessitates the development of new varieties with superior flavour quality. In this study, we characterised the volatile profiles of three new Okinawan pineapple breeding lines using gas chromatography (GC)–mass spectrometry (MS)‐based electronic nose (e‐nose), glycosylated volatiles, and alcohol‐acyltransferase (AAT) activity measurements. The volatile profiles of the fruits were selected using the GC–MS‐e‐nose and visualised using multivariate statistical plots. The glycosidically bound compounds were collected by solid‐phase extraction and hydrolysed by β‐glucosidase. In total, 12 hydroxyl‐group substances, including chavicol (medicinal‐herbal aroma), geraniol (floral‐sweet), and linalool (floral‐citrus), were released from these glycosides. Crude enzymes containing AAT were extracted, and their in vitro activities in 2‐methylbutyl acetate production were highly related to their contents. These results provide information regarding volatile profiles and bounded‐aroma resources, as well as the esterification potential of new Okinawan pineapple breeding lines for use in the agro‐food industry.

Fermentation with edible mushroom mycelia improves flavor characteristics and techno-functionalities of soybean protein

Fermentation has been considered as an effective strategy for improving the flavor characteristics and techno-functionalities of plant proteins. However, fermentation of soybean protein with mycelia of edible mushrooms has not been reported yet. In this study, edible mushrooms of Morchella Crassipes, Hedgehog Fungus, and Shiitake mushroom were selected for mycelium fermentation of soybean protein isolate (SPI). Mycelium fermentation modified the flavor profile of SPI in strain-dependent manner as indicated by electronic nose analysis. Sourness, bitterness, and astringency of SPI were altered after fermentation with increased proportion of sweet and tasteless amino acids. While, the proportion of bitter amino acids in SPI after 6 days fermentation with Morchella Crassipes (9.88%) was significant decreased as compared with that of untreated SPI (47.95%). Besides, off flavor of SPI was improved by fermentation as indicated by HS-SPME-GC-MS analysis with the relative content of hexanal be reduced from 22.40% to 6.93% after 6 days fermentation with Morchella Crassipes. Moreover, newly formed flavor compounds of benzaldehyde and 5-hydroxymethylfurfural have been identified after fermentation of SPI with mycelia of the selected edible mushroom. Fermentation with mycelium of Morchella Crassipes for six days significantly decreased surface hydrophobicity but increased sulfhydryl content in SPI with changed structural features. Those might contribute to the increased protein solubility (81.16 ± 2.77%) and foaming capacity (221.66 ± 2.35%), while the emulsifying capacity of SPI was significantly decreased. Findings gained in this study would benefit the production of soybean protein with superior flavor and techno-functionalities for potential food applications.

Flavour improvement in early generations of fresh market tomatoes (Solanum lycopersicum): I. Identification of QTL for sensory attributes, physicochemical measurements and volatile compounds

AbstractHuman sensory analysis is the most appropriate method for assessing the flavour of fresh market tomatoes, but it is very labour and time consuming. Therefore, sensory attributes are often neglected in early generations of breeding programmes and genetic studies, although there is a demand for tomatoes with improved flavour. In this study, the recently developed Breeders' Sensory Test was applied to an F2 mapping population derived from two parents with superior flavour. Sensory attributes, physicochemical measurements, volatiles and fruit weight were assessed in organic low‐input and hydroponic cultivation. A linkage map spanning 1070 cM was developed. In total, 71 quantitative trait loci (QTL) were detected for the means of both cultivation systems, 61 for organic and 46 for hydroponic cultivation. A proportion of 27% of the loci were co‐localized between both cultivation systems. Nine distinct QTL clusters for flavour‐related traits were identified, including a large cluster on chromosome 6 comprising five sensory and nine volatile QTL. The sensory QTL on chromosomes 2, 5, 6, 10 and 11, partly within clusters, are recommended for marker‐assisted selection.

Consumer Acceptance Studies of Margarine to Guide Product Development in the Food Industry.

Margarine exhibits significant variations in composition, allowing it to cater to diverse consumer segments. This study aimed to characterize the physical and sensory attributes of margarine samples available in the Brazilian market. Twelve commercial samples from six different brands, encompassing 30% to 80% of lipid contents, were subjected to instrumental texture analysis and affective assessment. A total of 112 consumers participated in acceptance tests and Check-All-That-Apply (CATA) evaluations, while another group of 62 subjects performed Projective Mapping. Samples with lipid percentages exceeding 70% achieved the highest average acceptance scores for taste and overall impression. The brand with the lowest lipid content (30%) exhibited a stronger association with negative attributes, including rancid flavor and aroma, bitterness, and metallic flavor, resulting in lower average scores for aroma, flavor, texture, and overall impression. However, these scores were not statistically different from samples with 50% and 60% lipid content. Reducing lipid levels in fat-based products such as margarine poses a challenge to food manufacturers, as consumers generally perceive higher lipid percentages as indicative of superior flavor quality.

Flavor improvement of fermented soybean foods by co-fermentation with Bacillus velezensis and Lactiplantibacillus plantarum

This study aimed to improve the flavor of fermented soybean foods and to investigate the molecular mechanisms of this flavor improvement phenomenon. Three fermented soybean foods (Bv, BvLp and Lp) were obtained using Bacillus velezensis SDL1 and Lactiplantibacillus plantarum Ly8 as starter cultures. Organoleptic evaluation showed that BvLp had the best flavor, followed by Bv and Lp. Next, some flavor-related physicochemical properties were investigated. Acid production level was ranked as Lp > BvLp > Bv, in which acetic acid and lactic acid were the main contributors to the variation in sample acid content, and BvLp contained the most lactic acid and isovaleric acid, 3.72−49.62 and 3.61−37.75 times those of Lp and Bv, respectively. Free amino acid analysis showed that the concentration trends of branched-chain amino acids and threonine were consistent with the sensory evaluation. Volatile metabolomics analysis indicated that acetoin and pyrazines were the key compounds for the superior flavor of BvLp. Finally, transcriptomic analysis, combined with the above data, revealed that under a parasitic-like interaction in microbial mixed culture, BvLp had significantly more flavor than Bv and Lp by reducing the acid content and increasing the concentration of some esters, acetoin and pyrazines.

SENSORY EVALUATION OF GAYO PEABERRY ARABICA COFFEE BY USING THE AHP METHOD

As other peaberrys, Gayo peaberry arabica coffee is very attractive because it has single larger and rounder seed. It is believed that the peaberry has superior flavor profile such as more fragrant aroma, stronger flavor, and denser than other arabica coffees. The purpose of this study was to evaluate the Gayo peaberry arabica coffee by using the analytical hierarchy process (AHP) method. Three varieties of Gayo peaberry arabica coffee were investigated i.e., Tim-tim, Bor-bor, and P88, while the processing method used were semi-wash and fully-wash methods. The sensory parameters observed included fragrance, flavor, acidity, body, and sweetness. The results showed that flavor (0.338) was the most important criterion than other taste criteria (fragrance 0.241; sweetness 0.196; acidity 0.115; and body 0.111). The variety that had the highest product acceptance rate was the Tim-tim variety processed by a full-wash method with a value of 0.203. The overall result of data analysis was acceptable because it had a consistency ratio below 0.1 (10%).

Analysis of physicochemical characteristics, antioxidant activity, and key aroma compounds of five flat peach cultivars grown in Xinjiang

Flat peach is one of the famous and characteristic table fruits in Xinjiang, China, and has attracted an increasing favorite because of its high nutrition and superior flavor. To evaluate the sensory properties and nutritional value of flat peach, functional components, antioxidant capacity, and aromatic compounds of five flat peach cultivars were measured by using high-performance liquid chromatography (HPLC) and headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS). The results indicated that sucrose, malic acid, and chlorogenic acid were the dominant sugars, organic acids, and phenolic compounds in flat peaches, respectively. The antioxidant activity ranged from 62 to 505.46 μmol TEs 100 g−1 FW. In addition, multivariate data analysis was used to discern differences in volatile compounds among different flat peach varieties, and 11 key aroma components were identified in combination with OAV analysis. These key aroma compounds interacted to present distinct green, fruity, floral, milk-like, sweet, and peach-like attributes and could be used as characteristic markers to distinguish the aroma of flat peach from other fruits. These results will provide a comprehensive chemical characterization of the edible quality, nutritional composition, and sensory properties of the five flat peach cultivars.

Individual and synergistic effect of multi-frequency ultrasound and electro-infrared pretreatments on polyphenol accumulation and drying characteristics of edible roses

The freeze-dried (FD) edible roses with high content of bioactive substances and superior flavor have been favored by consumers. Nevertheless, the development of freeze-dried rose industry has been plagued by a long drying time and low efficiency. This study investigated the effects of ultrasonic pretreatment (UP) in multi-frequency modes and electro-infrared pretreatment (EIP) prior to FD on polyphenol accumulation and drying characteristics of roses. The mechanism was explored by the changes in microstructure, equivalent circuit parameters, and phenol identifications of rose. The results showed that the FD time of roses decreased by 26 % after ultrasonic-infrared sequential synergistic pretreatment (UP + EIP) due to the damage of cell membrane permeability from UP. The quality attributes of UP + EIP products including color, phenols, and antioxidant activity (DPPH and ABTS radical scavenging rates) remarkably improved. UP + EIP significantly (p < 0.05) increased the content of polyphenols, namely quercetin-3β-d-glucoside, phlorizin, procyanidin B2, gallicacid, and rutin in the FD roses quantified by ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-qTOF-MS/MS). Therefore, UP + EIP is an effective pretreatment method for shortening FD time and producing high-quality FD rose products with enhanced polyphenol content.

Collection of native Theobroma cacao L. accessions from the Ecuadorian Amazon highlights a hotspot of cocoa diversity

Societal Impact Statement“Fine flavor” cocoa, known for its superior flavor and aroma, commands a higher price for farmers than “bulk” cocoa produced for market. These fine flavor cocoa varieties make an important contribution to the agricultural sector in Ecuador. However, cocoa diversity is threatened by deforestation. The effective preservation, characterization, and use of cocoa tree diversity are therefore essential to the future development of this market. We carried out participatory collection surveys with local communities in the Ecuadorian Amazon Forest, to evaluate the genetic diversity of native cocoa trees and protect trees as a resource for local communities. Accessing this wealth of diversity will aid farmers to safeguard cocoa against climate change and develop new varieties for market.Summary The aromatic Nacional variety of Theobroma cacao, emblematic of Ecuador, is highly sought after by the chocolate industry. The modern Nacional is a hybrid population resulting from genetic admixture that has lost the specificity of the ancestral variety. In the context of progressive forest disappearance, the objective of our study was to collect, safeguard, and evaluate native aromatic cocoa trees from Ecuadorian Amazon areas previously identified as areas of origin of the Nacional variety, as well as those that extend toward the northern Amazon. Four collection expeditions were organized in the Ecuadorian Amazon provinces of Zamora‐Chinchipe, Morona‐Santiago, and Pastaza in close collaboration with local communities. A total of 283 native accessions were collected and safeguarded at experimental stations and in local communities. The genetic diversity of the cocoa trees was analyzed by comparison to known genetic groups with a set of 48 simple sequence repeat (SSR) markers. This new collection clearly enriches the currently known diversity and improves knowledge of the global genetic structure of cocoa trees. Our results clarify the geographic origin of the Nacional variety in the vicinity of an archeological site that housed a Maya Chinchipe population that consumed cacao 5,000 years ago. In addition, our analyses revealed clues to the origin of Criollo, another ancient variety with a fine flavor. These new genetic resources will be used in breeding programs for the varietal improvement of new aromatic cocoa varieties and more globally for the selection of new varieties adapted to environmental changes.

A multi-omics framework reveals strawberry flavor genes and their regulatory elements.

Flavor is essential to consumer preference of foods and is an increasing focus of plant breeding programs. In fruit crops, identifying genes underlying volatile organic compounds has great promise to accelerate flavor improvement, but polyploidy and heterozygosity in many species have slowed progress. Here we use octoploid cultivated strawberry to demonstrate how genomic heterozygosity, transcriptomic intricacy and fruit metabolomic diversity can be treated as strengths and leveraged to uncover fruit flavor genes and their regulatory elements. Multi-omics datasets were generated including an expression quantitative trait loci map with 196 diverse breeding lines, haplotype-phased genomes of a highly-flavored breeding selection, a genome-wide structural variant map using five haplotypes, and volatile genome-wide association study (GWAS) with &gt; 300 individuals. Overlaying regulatory elements, structural variants and GWAS-linked allele-specific expression of numerous genes to variation in volatile compounds important to flavor. In one example, the functional role of anthranilate synthase alpha subunit 1 in methyl anthranilate biosynthesis was supported via fruit transient gene expression assays. These results demonstrate a framework for flavor gene discovery in fruit crops and a pathway to molecular breeding of cultivars with complex and desirable flavor.