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Comparative Analysis of Juice Volatiles in Selected Mandarins, Mandarin Relatives and Other Citrus Genotypes

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Comparative Analysis of Juice Volatiles in Selected Mandarins, Mandarin Relatives and Other Citrus Genotypes Research Article Received: 16 February 2017 Revised: 14 July 2017 Accepted article published: 21 July 2017 Published online in Wiley Online Library: 6 September 2017 (wileyonlinelibrary.com) DOI 10.1002/jsfa.8563 Comparative analysis of juice volatiles in selected mandarins, mandarin relatives and other citrus genotypes Yuan Yu,a Jinhe Bai,b Chunxian Chen,c Anne Plotto,b Elizabeth A Baldwinb and Frederick G Gmittera* Abstract BACKGROUND: Citrus fruit flavor is an important attribute prioritized in variety improvement. The present study compared juice volatiles compositions from 13 selected citrus genotypes, including six mandarins (Citrus reticulata), three sour oranges (Citrus aurantium), one blood orange (Citrus sinensis), one lime (Citrus limonia), one Clementine (Citrus clementina) and one satsuma (Citrus unshiu). RESULTS: Large differences were observed with respect to volatile compositions among the citrus genotypes. ‘Goutou’ sour orange contained the greatest number of volatile compounds and the largest volatile production level. ‘Ponkan’ mandarin had the smallest number of volatiles and ‘Owari’ satsuma yielded the lowest volatile production level. ‘Goutou’ sour orange and ‘Moro’ blood orange were clearly distinguished from other citrus genotypes based on the analysis of volatile compositions, even though they were assigned into one single group with two other sour oranges by the molecular marker profiles. CONCLUSIONS: The clustering analysis based on the aroma volatile compositions was able to differentiate mandarin varieties and natural sub-groups, and was also supported by the molecular marker study. The gas chromatography–mass spectrometry analysis of citrus juice aroma volatiles can be used as a tool to distinguish citrus genotypes and assist in the assessment of future citrus breeding programs. The aroma volatile profiles of the different citrus genotypes and inter-relationships detected among volatile compounds and among citrus genotypes will provide fundamental information on the development of marker-assisted selection in citrus breeding. © 2017 Society of Chemical Industry Supporting information may be found in the online version of this article. Keywords: aroma; fruit quality; lime; mandarin; sour orange; volatiles INTRODUCTION Taste, aroma, color and texture are the most important qual- Sweet oranges (Citrus sinensis), mandarins (or tangerines in the ity attributes of citrus fruits. Citrus aroma constituents are a mix- USA, Citrus reticulata) and grapefruit (Citrus paradisi)produced ture of monoterpenes, sesquiterpenes, alcohols, aldehydes, acids, 2,4,5 in Florida are important commodities in the US citrus market. esters, ketones and other volatiles. The aroma volatiles of sweet The production and consumption of mandarin fruits have been orange and grapefruit have been extensively studied because they are major commercial juice sources. Over 300 aroma volatiles increasing continuously in the USA as a result of their desirable have been reported for fresh orange juice.5 Several studies have attributes relating to a pleasant aroma and flavor, ease of peel- been performed on aroma volatiles in mandarin peel oil and ing, and high vitamin C, flavonoids and carotenoids. Awareness of essence,6–12 as well as in fresh fruits,13–18 although most of them the human health associated benefits derived from phytonutrients have studied only one or a few genotypes. Tietel et al.4 summa- is becoming a driver of consumer purchases and the consump- rized 37 consensus aroma volatiles in mandarin juice and nine tion of fruits.1,2 Traditional citrus breeding based on crossing and selection tends to be costly because hybrids usually take ≥5years before fruiting, and the capacity to grow hybrid populations, which ∗ is essential for selection of superior individuals, is restricted by Correspondence to: FG Gmitter, Citrus Research and Education Center, Univer- sity of Florida, Lake Alfred, FL, USA. E-mail: fgmitter@ufl.edu the large plant size (i.e. a tree crop). Molecular marker-assisted selection (MAS), which can help cull poor performing progeny a Citrus Research and Education Center, University of Florida, Lake Alfred, FL, USA seedlings at an early stage, could be a potentially useful solution to b Horticultural Research Laboratory, ARS, USDA, Fort Pierce, FL, USA these challenges in citrus breeding.3 The identification of molec- 1124 ular markers linked to citrus fruit taste and aroma can facilitate c Southeastern Fruit and Tree Nut Research Laboratory, ARS, USDA, Byron, GA, genetic improvement and the release of new superior varieties. USA J Sci Food Agric 2018; 98: 1124–1131 www.soci.org © 2017 Society of Chemical Industry Comparative analysis of juice volatiles www.soci.org of them were considered to be mandarin core aroma volatiles. as the Hunter color space value: L (black to white), a (green to Kerbiriou et al.19 detected 225 volatile compounds in juice sam- red) and b (blue to yellow), as well as the a over b ratio (indicating ples of 56 mandarin hybrids from the University of Florida Cit- orange color). Then, the 15 fruits from each genotype were ran- rus Research and Education Center (UF-CREC) citrus breeding pro- domly assigned into three pools with five fruits per pool, and each gram, and found that hybrids of an orange parentage were rich pool served as a biological replicate. Citrus fruits were manually in esters. Another study on 20 mandarin hybrids was performed peeled and gently juiced using a Hamilton Beach 932 manual bythesameteam,20 where mandarin hybrids produced volatiles citrus juicer (Hamilton Beach, Glen Allen, VA, USA) to prevent peel in fewer number and lower amounts, although with more alde- oil from entering the juice. Juice samples were used to collect hydes, whereas hybrids with sweet orange genetic background the data on the number of seeds, juice percentage (mL 100 g–1), generated more sesquiterpenes and esters. In addition, a follow-up juice color, soluble solids content (SSC) (g 100 mL–1) and titratable olfactometry study21 identified 49 aroma active compounds in acidity (TA) (g 100 mL–1, presented as a percentage of citric acid). five mandarin hybrids. Those results were confirmed by a recent comparative study22 that detected more volatiles, including more Volatile extraction and gas chromatography (GC)-mass sesquiterpenes and esters, in ‘Temple’ and ‘Ortanique’ tangors (C. spectrometry (MS) analysis reticulata × C. sinensis) compared to common mandarins such as For this, 3 mL of juice, 3 mL of saturated sodium chloride solution ‘Ora’ and ‘Ponkan’. The volatile profile was able to differentiate (359 g L–1)and6L of 1000 ppm 3-hexanone as an internal stan- four varieties, including mandarins, orange and pummelo (Cit- dard were mixed and placed in a 20-mL glass vial and capped rus maxima),23 and the experiment on two mandarin-pummelo with a magnetic crimp cap with a silicone/polytetrafluoroethylene hybrids revealed transgressive behaviour of aroma volatile pro- septum (Gerstel Inc., Linthicum, MD, USA). The vials were stored duction compared to their parents.24 at –20 ∘C, and then thawed under tap water and loaded into an Overall, these studies and others identified aroma volatiles in auto-sampler (Model MPS2; Gerstel Inc.) equipped with a cool- mandarin juice, and comparative analyses of aroma volatiles were ing tray holder (Laird Tech, Gothenburg, Sweden) controlled by a performed with respect to mandarins and other Citrus family mem- Peltier Thermostat (CTC Analytics AG, Zwingen, Switzerland). Sam- ∘ bers, such as sweet orange and pummelo. In the present study, ples were usually held for 2–20 h at 4 C until analyzed. For analy- ∘ we investigated aroma volatiles in 13 mandarins, mandarin rel- sis, the juice samples were incubated at 40 C for 30 min, and then a atives and other citrus genotypes, including sour orange and 2-cm tri-phase solid phase microextraction (SPME) fiber (50/30 m, lime, which have not previously been reported in exhaustive juice DVB/CAR/PDMS; Supelco, Bellefonte, PA, USA) was exposed to the ∘ aroma volatile studies. Meanwhile, we genotyped all the 13 cit- headspace for 1 h at 40 C. After exposure, the SPME fiber was rus selections in the University of Florida breeding program with a injected into the gas chromatograph (Model 6890; Agilent Tech- 1536-single nucleotide polymorphism (SNP) assay, which was used nologies, Santa Clara, CA, USA) injector port (splitless mode) to release the compounds for 15 min at 250 ∘C. The GC equipment to genotype a mandarin F1 population and construct mandarin genetic linkage maps.25 We used different analysis tools aiming and settings were: DB-5 columns (length 60 m, inner diameter to understand relationships among different genotypes of man- 0.25 mm, film thickness 1.00 m; J&W Scientific, Folsom, CA, USA), darins, mandarin relatives and other citrus genotypes, in accor- coupled with a 5973 N MS detector (Agilent Technologies). The col- ∘ –1 dance with their aroma volatile profile and molecular marker geno- umn oven was programmed to increase at 4 Cmin from the ∘ ∘ ∘ –1 ∘ typic data. The research would provide fundamental information initial 40 C to 230 C, and then ramped at 100 Cmin to 260 C on development of MAS for citrus fruit quality improvement. and held for 11.70 min for a total run time of 60 min. Helium was used as carrier gas at a flow rate of 1.5 mL min−1. Inlet, ionizing source and transfer line were kept at 250, 230 and 280 ∘C, respec- MATERIALS AND METHODS tively. Mass units were monitored from 30 to 250 m/z and ionized at 70 eV. Data were collected with the ChemStation G1701AA data Plant material system (Hewlett-Packard, Palo Alto, CA, USA). A mixture of C-5 to Thirteen citrus genotypes were studied, including six mandarins C-18 n-alkanes was run at the beginning of each day separately to (C. reticulata cv.‘Fortune’, ‘King’. ‘Murcott’, ‘Pimpled’, ‘Ponkan’ and calculate retention indices (RI).26 ‘Sunki’), three sour oranges (Citrus aurantium cv. ‘Goutou’, ‘Zhu- luan’ and an unknown named ‘Sour’), one blood orange (C.
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