Strawberry Volatiles Research Articles

Determination of volatile profiles of woodland strawberry (Fragaria vesca) during fruit maturation by HS-SPME GC-MS.

Aroma is an important agronomic trait for strawberries, and the improvement of fruit flavor is a key goal in current strawberry breeding programs. Fragaria vesca (also known as woodland strawberry) has become an excellent model plant with exquisite flavor, a small genome size, and a short life cycle. Thus, the comprehensive identification of fruit volatiles and their accumulation pattern of F. vesca strawberries are very important and necessary to the fruit aroma study. This study examined the volatile profile changes from the fruits of three F. vesca genotypes during maturation using the headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME GC-MS) with multivariate analysis. A total of 191 putative volatile compounds were identified, while 152, 159, and 175 volatiles were detected in 20-30 dap fruits of Hawaii 4 (HW), Reugen (RG), and Yellow Wonder (YW), respectively. Aldehydes and alcohols predominated in the early time point while esters were predominant during the late time point. Ketones were the dominant compounds from F. vesca strawberries fruits at the ripe stage. Certain genotype-characteristic volatiles were identified, including eugenol, γ-octalactone, and δ-decalactone were only detected in YW, and mesifurane was found in HW. RG and YW showed very similar volatile compositions, but YW presented a greater number of volatiles and RG yielded a higher content. Differences in the volatile composition may be primarily due to genetic relationships. The metabolic changes that occurred during fruit ripening and characteristic volatiles will be a useful reference for future strawberry volatile studies. This article is protected by copyright. All rights reserved.

Changes in Volatile Compounds in Short-Term High CO2-Treated 'Seolhyang' Strawberry (Fragaria × ananassa) Fruit during Cold Storage.

‘Seolhyang’ strawberry is harvested before it is fully ripened and treated with CO2 to extend the shelf-life. However, the volatile changes in the ‘Seolhyang’ strawberry after short-term CO2 treatment have not been investigated, although the volatile profile is an important quality attribute. Herein, we investigated the effect of short-term high CO2 treatment on the changes in the composition of volatile compounds in ‘Seolhyang’ strawberries at two ripening stages (i.e., half-red and bright-red) during cold storage using headspace solid-phase microextraction and gas chromatography-mass spectrometry. Furthermore, the effect of CO2 treatment on fruit quality with respect to the aroma was investigated. A total of 30 volatile compounds were identified. Storage increased the volatile compound concentrations, and the total concentration of volatiles in the CO2-treated strawberries was lower than that of the untreated strawberries during storage. The production of some characteristic strawberry volatiles (e.g., 4-methoxy-2,5-dimethyl-3(2H)-furanone) was inhibited in CO2-treated strawberries. However, CO2 treatment helped maintain the concentrations of hexanal and 2-hexenal, which are responsible for the fresh odor in strawberries. Interestingly, CO2 treatment suppressed the production of off-odor volatiles, acetaldehyde, and hexanoic acid during strawberry storage. Thus, short-term CO2 treatment may help maintain the fresh aroma of strawberries during cold storage.

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 > 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.

Strawberry fruit FanCXE1 carboxylesterase is involved in the catabolism of volatile esters during the ripening process.

Volatile compounds produced during ripening of strawberry are key determinants of fruit quality and consumer preference. Strawberry volatiles are largely esters which are synthesized by alcohol acyltransferases (AATs) and degraded by carboxylesterases (CXEs). Although CXE activity can have a marked influence on volatile contents in ripe strawberry fruits, CXE function and regulation in them are poorly known. Here, we report the biochemical and functional characterization of the fruit receptacle-specific and ripening-related carboxylesterase FanCXE1. The expression of the corresponding gene was found to be antagonistically regulated by auxins and abscisic acid, key hormones that regulate fruit growth and ripening in strawberry. In vitro, FanCXE1 was able to hydrolyze artificial ester substrates similar to those produced by ripe strawberry fruits. Transient suppression of the FanCXE1 gene by RNAi resulted in an increase of important volatile esters such as methyl hexanoate, methyl butanoate and ethyl hexanoate as well as a decrease of the alcohols hexenol and linanool. The results of this work enhance our understanding of the molecular basis for volatile syntheses and facilitate production of better flavored strawberry fruits by introduction of the relevant alleles into common cultivars.

Quality Properties and Headspace Volatiles of Hot Air-Dried Strawberries.

Strawberry is one of the most important soft fruit in food industry. With its flavor and antioxidant content, it is widely used mainly in confectionery and beverage industry. Strawberries are perishable and this limits their usage in foods. For that reason, drying is one of the solutions to make them durable. The aim of this study is to analyze certain quality parameters and headspace volatiles of hot air-dried 'Florida Fortuna' and 'Osmanlı' strawberries (Fragaria xananassa) at 50, 60 and 70°C. The results showed that drying the strawberry samples at high drying temperature caused a decrease in acidity and bulk density and a deterioration in ascorbic acid content. However, total phenolic contents and total color difference values of both of strawberry samples increased with drying process. A total of 24 headspace volatiles were detected in dried strawberries. The most abundant volatiles were dimethyl sulfide, acetic acid and acetone. The highest number of compound groups were aldehydes, acids, esters and sugar degradation products, respectively. The highest retention of fresh strawberry volatiles and minimum formation of sugar degradation products were obtained by drying at low temperature.

Effect of hydrocolloids, sugar, and citric acid on strawberry volatiles in a gummy candy

Effect of hydrocolloids, sugar, and citric acid on strawberry volatiles in a gummy candy

Quantitative Analysis of Fragrance and Odorants Released from Fresh and Decaying Strawberries

The classes and concentrations of volatile organic compounds (VOC) released from fresh and decaying strawberries were investigated and compared. In this study, a total of 147 strawberry volatiles were quantified before and after nine days of storage to explore differences in the aroma profile between fresh strawberries (storage days (SRD) of 0, 1, and 3) and those that had started to decay (SRD = 6 and 9). In terms of concentration, seven compounds dominated the aroma profile of fresh strawberries (relative composition (RC) up to 97.4% by mass, sum concentration): (1) ethyl acetate = 518 mg·m−3, (2) methyl acetate = 239 mg·m−3, (3) ethyl butyrate = 13.5 mg·m−3, (4) methyl butyrate = 11.1 mg·m−3, (5) acetaldehyde = 24.9 mg·m−3, (6) acetic acid = 15.2 mg·m−3, and (7) acetone = 13.9 mg·m−3. In contrast, two alcohols dominated the aroma profile of decayed samples (RC up to 98.6%): (1) ethyl alcohol = 94.2 mg·m−3 and (2) isobutyl alcohol = 289 mg·m−3. Alternatively; if the aroma profiles are re-evaluated by summing odor activity values (ΣOAV); four ester compounds ((1) ethyl butyrate (6,160); (2) ethyl hexanoate (3,608); (3) ethyl isovalerate (1,592); and (4) ethyl 2-methylbutyrate (942)) were identified as the key constituents of fresh strawberry aroma (SRD-0). As the strawberries began to decay; isobutyl alcohol recorded the maximum OAV of 114 (relative proportion (RP) (SRD = 6) = 58.3%). However, as the decay process continued, the total OAV dropped further by 3 to 4 orders of magnitude—decreasing to 196 on SRD = 6 to 7.37 on SRD = 9. The overall results of this study confirm dramatic changes in the aroma profile of strawberries over time, especially with the onset of decay.

Chemical characterisation of two Australian-grown strawberry varieties by using comprehensive two-dimensional gas chromatography–mass spectrometry

The volatile compositions of two strawberry varieties (‘Albion’ and ‘Juliette’) grown in Australia were analysed with comprehensive two-dimensional gas chromatography (GC×GC) combined with time-of-flight mass spectrometry (ToFMS). A suitable data processing method was created that should be sufficiently robust to compare sample chromatograms within a harvest season as well as between seasons with relative ease. Ninety-four compounds were tentatively or positively identified between the two varieties of which 20 have apparently not been previously reported to be components of strawberry volatiles. It was determined that Albion and Juliette have comparable total soluble solids content close to 11°Brix. However, since Juliette exhibited a lower titratable acidity content than Albion, it consequently has a slightly larger sugar/acid ratio. The presence of 2,5-dimethyl-4-hydroxy-(2H)-furan-3-one (furaneol) was detected only in Juliette and this variety also had a higher area percentage of 2,5-dimethyl-4-methoxy-(2H)-furan-3-one (mesifuran), according to the sampling protocol. These experimental results offer possible explanations as to why Juliette is perceived to have a sweeter flavour than Albion.

Effect of Enzymes on Strawberry Volatiles during Storage, at Different Ripeness Level, in Different Cultivars, and during Eating

Strawberry samples with enzyme activity and without enzyme activity (stannous chloride added) were measured for real-time formation of lipoxygenase (LOX) derived aroma compounds after 5 min pureeing using selected ion flow tube-mass spectrometry (SIFT-MS). The concentration of (Z)-3-hexenal and (E)-2-hexenal increased immediately after blending and gradually decreased over time, while hexanal concentration increased for at least 5 min in ground strawberries. The formation of hexanal was slower than the formation of (Z)-3-hexenal and (E)-2-hexenal in the headspace of pureed strawberries. The concentration of LOX aldehydes and esters significantly increased during refrigerated storage. Damaging strawberries increased the concentration of LOX aldehydes but did not significantly affect the concentration of esters. The concentrations of many of the esters were strongly correlated to their corresponded acids and/or aldehydes. The concentration of LOX-generated aldehydes decreased during ripening, while fruity esters increased. Different varieties had different aroma profiles and esters were the greatest percentage of the volatiles. The aroma release of some of the LOX-derived aldehydes in the mouthspace in whole strawberries compared to chopped strawberries showed that these volatiles are formed in the mouth during chewing. The persistence of LOX-derived compounds was higher than esters after swallowing. The mouthspace after and before swallowing persistence ratio of esters decreased as the chain length of the acid part of the ester compounds increased in whole strawberries. The storage studies showed that the concentrations of fruity and fresh volatiles increased during ripening and storage while damaging only increases the fresh volatiles. The nose, mouth, and headspace information can be used in the flavor industry to improve the formula of natural strawberry flavor by considering human perception during eating.

Analysis of strawberry volatiles using comprehensive two-dimensional gas chromatography with headspace solid-phase microextraction

The aims of the current study were to develop an enantioselective multi-dimensional gas chromatography (GC × GC) method for the examination of strawberry volatiles and to use this method to make comparisons between the volatile profiles of different cultivars and between fresh picked and post-harvest berries of the same cultivar. Strawberry volatiles were sampled using solid-phase microextraction (SPME), and the repeatability and reproducibility of this method was examined. Semi-quantitative analysis of the volatiles was conducted using the relatively new technique of comprehensive multi-dimensional gas chromatography, using enantioselective (chiral) columns for the differentiation of analyte enantiomers. Chiral GC × GC facilitated the detection of key enantiomers in strawberry flavour. The (−)-enantiomer of 2,5-dimethyl-4-hydroxy-(2 H)-furan-3-one (DMHF) and the S-enantiomer of linalool were tentatively identified as the predominant forms in both the cultivars Selva and Adina. The compounds benzaldehyde and methyl hexanoate were shown to decrease in post-harvest berries, whilst DMHF and nerolidol increased upon storage.

Isolation, cloning and expression of a multifunctional O-methyltransferase capable of forming 2,5-dimethyl-4-methoxy-3(2H)-furanone, one of the key aroma compounds in strawberry fruits.

Strawberry fruits contain an uncommon group of key aroma compounds with a 2,5-dimethyl-3(2H)-furanone structure. Here, we report on the methylation of 2,5-dimethyl-4-hydroxy-3(2H)-furanone (DMHF) to 2,5-dimethyl-4-methoxy-3(2H)-furanone (DMMF) by a S-adenosyl-L-methionine dependent O-methyltransferase, the cloning of the corresponding cDNA and characterization of the encoded protein. Northern-hybridization indicated that the Strawberry-OMT specific transcripts accumulated during ripening in strawberry fruits and were absent in root, petiole, leaf and flower. The protein was functionally expressed in E. coli and exhibited a substrate specificity for catechol, caffeic acid, protocatechuic aldehyde, caffeoyl CoA and DMHF. A common structural feature of the accepted substrates was a o-diphenolic structure also present in DMHF in its dienolic tautomer. FaOMT is active as a homodimer and the native enzyme shows optimum activity at pH 8.5 and 37 degrees C. It does not require a cofactor for enzymatic activity. Due to the expression pattern of FaOMT and the enzymatic activity in the different stages of fruit ripening we suppose that FaOMT is involved in lignification of the achenes and the vascular bundles in the expanding fruit. In addition, it is concluded that the Strawberry-OMT plays an important role in the biosynthesis of strawberry volatiles such as vanillin and DMMF.

Volatile compounds of selected strawberry varieties analyzed by purge-and-trap headspace GC-MS.

Six strawberry (Fragaria x ananassa Duch.) varieties, Senga Sengana, Jonsok, Polka, Korona, Bounty, and Honeoye, were studied in relation to composition of their volatile headspace compounds collected by dynamic purge-and-trap concentration. Also, the influence of cultivation technique on the volatile compounds was tested by comparing conventionally and organically cultivated strawberry varieties. In addition, the effect of geographical origin on strawberry volatiles was investigated. The emphasis of this study was lay in the industrial utilization of strawberry. A total of 52 different volatile compounds were determined from frozen strawberries grown in 1997 and 1998. In addition to individual compounds, total areas of chromatogram profiles were compared. The major esters were found to be methyl butanoate, ethyl butanoate, methyl hexanoate, and ethyl hexanoate. With the application of principal component analysis and analysis of variance to the data matrix, the varieties were classified into three groups. Var. Honeoye was most different from the others. Organic cultivation had no effect on strawberry volatiles.

Determination of strawberry volatiles with low resolution gas phase FT-IR analyser

A low-resolution gas phase Fourier transform infrared (FT-IR) analyser, designed for industrial and environmental applications, was applied to the analysis of volatile compounds of strawberry. The frozen fruit was thawed, mashed and transferred into a glass bottle mounted to a sampling system. Volatile compounds were collected by gently heating in a vacuum and then removed into the sample cell of the gas analyser at atmospheric pressure. The quantitative analysis of 14 compounds in all was performed by a built-in microcomputer applying multicomponent analysis. GC-MS applying dynamic headspace sampling was used as a reference method. A spectral library consisting of 24 typical aroma volatiles of strawberry was created before the analyses. The highest proportions were measured for acetone and 2,5-dimethyl-4-hydroxy-3(2H)-furanone. The average coefficient of variation (CV) of the relative amount of all the compounds was 35%, that of acetone being the lowest, 3.3%. Significant differences between strawberry varieties were noticed.

Identification of the SAAT gene involved in strawberry flavor biogenesis by use of DNA microarrays.

Fruit flavor is a result of a complex mixture of numerous compounds. The formation of these compounds is closely correlated with the metabolic changes occurring during fruit maturation. Here, we describe the use of DNA microarrays and appropriate statistical analyses to dissect a complex developmental process. In doing so, we have identified a novel strawberry alcohol acyltransferase (SAAT) gene that plays a crucial role in flavor biogenesis in ripening fruit. Volatile esters are quantitatively and qualitatively the most important compounds providing fruity odors. Biochemical evidence for involvement of the SAAT gene in formation of fruity esters is provided by characterizing the recombinant protein expressed in Escherichia coli. The SAAT enzyme showed maximum activity with aliphatic medium-chain alcohols, whose corresponding esters are major components of strawberry volatiles. The enzyme was capable of utilizing short- and medium-chain, branched, and aromatic acyl-CoA molecules as cosubstrates. The results suggest that the formation of volatile esters in fruit is subject to the availability of acyl-CoA molecules and alcohol substrates and is dictated by the temporal expression pattern of the SAAT gene(s) and substrate specificity of the SAAT enzyme(s).

The Influence of Sugar Concentrations on the Vapor Pressure of Food Odor Volatiles in Aqueous Solutions

SUMMARY— The addition of fructose or invert sugar in very high concentrations to aqueous solutions containing strawberry volatiles or synthetic compounds respectively, results for some components in a decrease of peak heights in gas chromatograms of the vapors over the solutions; the phenomenon is thought to be of interest for studying flavor retention in juice concentrates.

Volatiles from strawberries : II. Combined mass spectrometry and gas chromatography on complex mixtures

The technique of combined mass spectrometry and gas chromatography has been applied to the analysis of a complex oil of strawberry volatiles. Capillary gas chromatography indicated over 150 components. Most of the major components have been identified. These include alcohols, esters, acetals, aldehydes, furfural, methyl furfural, aromatic aldehydes, ketones, and esters, a few terpenes, and a few aromatic hydrocarbons. The strength of this technique is shown by the determination of several compounds in one sharp chromatographic peak. On the other hand certain prominent species could not bbe identified by the low resolution (resolution about 200) mass spectral data obtained.