Study of commercial quality parameters, sugars, phenolics, carotenoids and plastids in different tomato varieties

1 Study of commercial quality parameters, sugars, phenolics, carotenoids and plastids 1 in different tomato varieties 2 Elena Coyago-Cruzabc, Mireia Corellad, Alfonso Moriana ad, Paula Mapelli-Brahmb, 3 Dolores Hernanze, Carla M. Stincob, Elena Beltrán-Sinchiguanof, Antonio J. Meléndez-4 Martínezb* 5 a. Dpto. Ciencias Agroforestales, Universidad de Sevilla, Escuela Técnica Superior de 6 Ingeniería Agronómica, Carretera de Utrera Km 1, 41013 Sevilla, Spain. 7 b. Food Colour & Quality Lab., Area of Nutrition & Food Science, Universidad de 8 Sevilla, Facultad de Farmacia, 41012 Sevilla, Spain. 9 c. Carrera de Ingeniería en Biotecnología de los Recursos Naturales, Universidad 10 Politécnica Salesiana, Sede Quito, Campus El Girón, Av. 12 de Octubre, Quito, 11 Ecuador. 12 d. Unidad Asociada al CSIC de Uso sostenible del suelo y el agua en la agricultura 13 (US-IRNAS), Crta. de Utrera Km 1, 41013 Sevilla, Spain. 14 e. Dpto. Química Analítica, Universidad de Sevilla, Facultad de Farmacia, 41012 15 Sevilla, Spain. 16 f. Centro de Investigación de Alimentos, CIAL, Ingeniería de Alimentos, Facultad de 17 Ciencias de la Ingeniería e Industrias, Universidad Tecnológica Equinoccial, 18 EC171029 Quito, Ecuador. 19 CORRESPONDING AUTHOR 20 *Phone: +34-954557017 21 E-mail: [email protected] 22 Postprint of: Food Chemistry 277: 480-489 (2019) 2 EMAIL ADDRESSES FOR AUTHORS 23 Coyago-Cruz, Elena ecoyag[email protected] 24 Corell, Mireia [email protected] 25 Hernaz, Dolores [email protected] 26 Moriana, Alfonso [email protected] 27 Stinco, Carla [email protected] 28 Mapelli-Brahm, Paula [email protected] 29 Beltrán-Sinchiguano, Elena elena.be[email protected] 30 Meléndez-Martínez, Antonio [email protected] 31 3 Abstract 32 The aim of this study was to assess commercial quality parameters, sugars, phenolics, 33 carotenoids and plastid in diverse and little studied tomato varieties to gain insight into 34 their commercial and functional quality and reveal possible noticeable differences. Five 35 cherry tomato varieties and six common (i.e., non-cherry) tomatoes were evaluated. The 36 highest levels of lycopene were detected in ʽTigerellaʼ and ʽByelsaʼ, and those of phytoene 37 in ʽOrangeʾ, those of phenolics in ʽGreen Zebraʼ, all of them common tomatoes.The levels 38 of sugars in both groups of tomatoes were comparable. Interesting differences in plastid 39 carotenoid-accumulating sub-structures as a function of the carotenoid profile were 40 observed. Given the importance of chromoplasts in the deposition of carotenoids in plants 41 and their release during digestion, this information can be valuable in investigations on the 42 regulation of the biosynthesis and the bioavailability of tomato carotenoids. 43 44 Keywords: Functional foods; chromoplasts; phytoene; phytofluene; ultrastructure, 45 transmission electron microscopy (TEM) 46 4 1. INTRODUCTION 47 Tomato (Solanum lycopersicum L.) is one of the vegetables more consumed in the world 48 and the basis of other food products. They provide important compounds like sugars, 49 minerals, vitamins, carotenoids and phenolics, whose levels can vary markedly as a 50 function of genetics, physiological, agronomic, technological or other factors (Coyago51 Cruz, Corell, Stinco, et al., 2017; Antonio J. Meléndez-Martínez, Fraser, & Bramley, 52 2010). Given the economic and nutritional importance of tomato and derivatives, it is not 53 surprising that their study from different perspectives, including composition (Cichon, 54 Riedl, & Schwartz, 2017; R. M. Schweiggert & Carle, 2017), sustainable production 55 approaches (Borghesi et al., 2011; Coyago-Cruz et al., 2018; Coyago-Cruz, Corell, Stinco, 56 et al., 2017) release of components during digestion (Mapelli-Brahm, Corte-Real, 57 Meléndez-Martínez, & Bohn, 2017; Talens, Mora, Bramley, & Fraser, 2016) and possible 58 health benefits derived from their intake (Cooperstone et al., 2015), continue featuring in 59 the latest scientific literature. 60 Apart from weight, size and total soluble contents (related to sugar content), colour 61 is one of the key parameters evaluated in the context of commercial quality and food 62 acceptability. Although traditionally red tomatoes have been marketed and are usually 63 preferred by consumers; varieties with other colours (including green, yellow, orange, 64 purple) are not as commonly found in the market and have been less studied (Borghesi et 65 al., 2011; Cooperstone et al., 2017; Antonio J. Meléndez-Martínez et al., 2010; Yuan, Li, 66 & Wilson, 2008) . The red colour of tomatoes is mainly due to their carotenoid profile 67 while in darker varieties, this attribute can be due mainly to the retention of chlorophyll 68 and the accumulation of lycopene (Park, Sangwanangkul, & Baek, 2018) or even the 69 accumulation of both carotenoids and anthocyanins (Borghesi et al., 2011). In non-green 70 5 tomatoes, carotenoids are accumulated in a type of plastid named chromoplast, whose 71 biogenesis is associated with chlorophyll degradation (Li & Yuan, 2013). There are 72 different classes of chromoplasts with different carotenoid accumulating structures such 73 as,crystals or globules, among others, which depends on the carotenoid profile of the part 74 of the plant (root, fruit, petal, etc.) in question (R. M. Schweiggert & Carle, 2017). The 75 study of the types of chromoplasts is relevant as theyare key organelles for the deposition 76 of carotenoids and also important in relation to the release of carotenoids during digestion, 77 one of the key factors governing their bioavailability (R. M. Schweiggert & Carle, 2017). 78 Taking all these facts together, the goal of this study was to assess commercial 79 quality parameters (equatorial and longitudinal diameter, weight, soluble solids and 80 colour), sugars, phenolics and carotenoids contents as well as chromoplast morphology in 81 diverse and little studied tomato varieties in order to gain further insight into their 82 commercial and functional quality and reveal possible noticeable differences. 83 2. MATERIALS AND METHODS 84 2.1 Reagents and standards 85 Analytical grade reagents, specifically methanol (PumChem CID: 887), trichloromethane 86 (PumChem CID: 6212) and hydrochloric acid (PunChem CID: 313) were purchased from 87 Labscan (Dublin, Ireland). HPLC grade reagents, like methanol, acetonitrile (PumChem 88 CID: 6342) and ethyl acetate (PumChem CID: 8857) were obtained from Panreac 89 (Barcelona, Spain). Ultra-pure water was obtained by means of a NANOpure DlamondTM 90 system (Barnsted Inc., Dubuque, IO). Lutein, lycopene, phytoene and phytofluene were 91 obtained from appropriate sources as described elsewhere (Melendez-Martinez, Stinco, 92 Liu, & Wang, 2013; Antonio J. Meléndez-Martínez, Vicario, & Heredia, 2007), β-carotene 93 (PumChem CID: 5280489) was purchased from Sigma-Aldrich (Taukirchen, Germany), 94 6 and quercetin (PumChem CID: 370), ferulic acid, caffeic acid, p-coumaric acid (PumChem 95 CID: 637542), and gallic acid (PumChem CID: 1794427) were from Sigma-Aldrich 96 (Madrid, Spain). Glutaraldehyde, formaldehyde and buffer sodium cacodylate were 97 acquired from Ted Pella, Inc. (Redding, USA). 98 99 2.2. Plant materials 100 Eleven tomato (Solanum lycopersicum L.) varieties were studied. Five cherry varieties of 101 Granada La Palma Company, i.e. ‘Cherry amarillo’ (A), ʽCherry pera clásicoʼ (B), ʽCherry 102 pera naranjaʼ (C) and ʽMinichocmato peraʼ (D) (corresponding to 4 Mixcherrys) and 103 ʽCherry cerejaʼ (E), were selected and obtained from a local market in Sevilla. ʽCherry 104 amarilloʼ and ʽCherry cerejaʼ were round varieties with yellow and red colour, 105 respectively, while ʽCherry pera clásicoʼ, ʽCherry pera naranjaʼ and ʽMinichocmato peraʼ 106 were pear varieties with red, orange and green-red colour, respectively. Forty fruits of each 107 cherry variety were considered for the commercial quality analyses. On the other hand, six 108 “common” (that is, non-cherry) tomato varieties, i.e. ʽGreen Zebraʼ(F), ʽSunchocolaʼ(G), 109 ʽTigerellaʼ (H), ʽByelsaʼ (I), ʽPalamósʼ (J), and ʽOrangeʼ (K), were grown in a greenhouse 110 at Escuela Técnica Superior de Ingeniería Agronómica (E.T.S.I.A.) of Universidad de 111 Sevilla (Sevilla, South Spain, 37º21'09.71'' Lat. N, 5º56'19.13'' Long. W, 33 m a.s.l.) during 112 spring of 2015 (23rd February to 15th June), exceptʽSunchocolaʼ, which was grown during 113 autumn of 2015 (23rd September to 15th December). The seeds of the varieties ʽByelsaʼ and 114 ʽPalamósʼ were provided by Fitó (Almería, Spain), ʽSunchocolaʼ and ʽOrangeʼ or ʽOrange 115 Wellingtonʼ by W. Atlee Burpee (Warminster, USA) and ʽGreen Zebraʼ and ʽTigerellaʼ by 116 Magic Garden Seeds (Regensburg, Germany). ʽGreen Zebraʼ and ʽTigerellaʼ were striped 117 round medium to small varieties with green-yellow and red-yellow colour, respectively. 118 ʽSunchocolaʼ is a round medium to small variety, which has a green-red colour. ʽByelsaʼ 119 7 and ʽPalamósʼ are red medium to large tomatoes, with a pear and round form, respectively. 120 ʽOrangeʼ is a very large variety with orange colour. Three ripe fruits of seven plants (21 121 samples of tomato) of each common tomato variety were sampled for the analyses of 122 commercial quality. The optimum degree of maturity for harvesting was determined 123 visually by considering their colour. 124 The measurements of size, weight, soluble solids, humidity, and colour as well as the 125 microscopic analyses were performed on the fresh fruit. Afterwards, the seeds and inside 126 locular tissues were removed and the remaining parts of the fruits of each variety were 127 mixed. Afterwards, the mixtures were divided into two halves, which were ground in a 128 basic A 11 IKA mill, frozen at -80°C and freeze-dried (Cryodos system). The freeze-dried 129 samples were stored under a nitrogen atmosphere in dark glass bottles hermetically sealed. 130 These were kept at -21 °C until the analyses. 131 2.3. Commercial quality assessments 132 Equatorial and longitudinal diameter (cm), weight (g), soluble solids (° Brix), humidity and 133 colour were measured on the fresh tomatoes. Analyses were performed with 40 replicates 134 for the cherry varieties and 21 replicates for the common varieties. The soluble solids (SS) 135 were quantified with a Hand-Refractometer RHC-200ATC (Huake, China) using a drop of 136 tomato juice. The colour parameters corresponding to the uniform colour space CIELAB 137 (L*, a*, b*, C*ab and hab) were obtained directly from a CM-700d colorimeter (Minolta, 138 Japan) as described elsewhere Coyago-Cruz et al. (2017). 139 2.4 Analysis of sugars, phenolic compounds and carotenoids 140 2.4.1 Analysis of sugars 141 Sugars were extracted and analyzed as described by Kasim & Kasim, (2015) with 142 slight modifications. The two homogenized freeze-dried powder were extracted in 143 triplicate. Approximately 200 mg of the freeze-dried sample was extracted with 5 mL of 144 8 water. The mixture was vortexed, sonicated for 5 min, and centrifuged at 4190 g for 7 min 145 at 4 ºC. The extracts were filtered through Millipore membranes (0.45 µm pore, 15 mm 146 diameter) (Agilent Technologies, Spain) prior to their injection in the HPLC system. All 147 the extracts were injected twice. The HPLC analyses were carried out on an Agilent 1200 148 chromatograph equipped with a RID-detector (Agilent Technologies, Palo Alto, CA. USA) 149 and a Zorbax Carbohydrate column (4.6 mm × 150 mm) kept at 30 ºC. The injection 150 volume was 5 μL and the flow rate was 1 mL/min. The mobile phase consisted of 151 acetonitrile/water (70:30). The open lab ChemStation software was used. Sugars were 152 identified by comparing their retention time with those of standards. Fructose, glucose and 153 sucrose were identified with standards by comparing their retention times and the total 154 sugar content (TSC) were calculated as the sum of individual sugars. 155 2.4.2 Analysis of phenolic compounds 156 The extractions and analyses were carried out as described by Coyago-Cruz, et al. (2017). 157 The two homogenized freeze-dried powder were extracted in triplicate. Briefly, 158 approximately 0.5 g of homogenized freeze-dried powder was vortexed and sonicated for 159 15 min with 15 mL of 75% aqueous methanol (v/v) containing HCl 0.1% (v/v). The 160 mixture was centrifuged at 4190 g for 7 min at 4 ºC; the supernatant was collected and the 161 residue subjected to the same process twice, using only 5 mL of aqueous methanol. The 162 extract was stored at - 20 °C until analysis. The extracts were filtered through Millipore 163 membranes (0.45 µm pore, 15 mm diameter) (Agilent Technologies, Spain) for injection in 164 the UHPLC system. One mL of the extract obtained was dissolved in 4 mL of 0.01% 165 formic acid in water or injection in the UHPLC system. All the extracts were injected 166 twice. The UHPLC analyses were carried out on an Agilent 1290 chromatograph equipped 167 with a diode-array detector (Agilent Technologies, Palo Alto, CA. USA) and an Eclipse 168 Plus C18 column (1.8 um, 2.1 × 5 mm) at 30 °C. The mobile phase consisted of 1 mL/min 169 9 of 0.01% of formic acid in water (solvent A) and acetonitrile (solvent B) with the linear 170 gradient elution: 100% A, 0 min; 95% A + 5% B + 20% C, 5 min; 50% A + 50% B, 20 171 min; washing and re-balancing of the column, 22 min. The open lab ChemStation software 172 was used and the chromatograms were monitored at 280, 320 and 370 nm for the 173 quantification of p-hydroxybenzoic acid, p-coumaric acid, caffeic acid, chlorogenic acid, 174 gallic acid, ferulic acid, naringin, crisin, quercetrin and quercetin, respectively. Phenolics 175 were identified with standards by comparing their retention time and UV-vis spectra. Total 176 phenolic content was calculated as the sum of individual phenolics. 177 2.4.3 Analysis of carotenoids 178 Carotenoids were extracted and analyzed as described by Coyago-Cruz, et al. (2017). The 179 two homogenized freeze-dried powder samples were extracted in triplicate. In brief, 180 approximately 20 mg of homogenized freeze-dried powder were mixed with 250 μL of 181 methanol, 500 μL of trichloromethane and 250 μL of Milli-Q water. The coloured organic 182 fractions were evaporated and stored under a nitrogen atmosphere at -20 °C until the 183 chromatographic analysis. The dry residue was re-dissolved in 40 µL of ethyl acetate prior 184 to their injection in the RRLC system. All the extracts were injected twice. These were 185 carried out on an Agilent 1260 system equipped with a diode-array detector. A C18 186 Poroshell 120 column (2.7 μm, 5 cm x 4.6 mm) (Agilent, Palo Alto, CA) at 30 ºC was used 187 for the separations. The mobile phase consisted of 1 mL/min of acetonitrile (solvent A), 188 methanol (solvent B) and ethyl acetate (solvent C) with the linear gradient elution: 85% A 189 + 15% B, 0 min; 60% A + 20% B + 20% C, 5 min; 60 % A + 20% B + 20% C, 7 min; 85% 190 A + 15% B, 9 min; 85% A + 15 % B, 12 min. The open lab ChemStation software was 191 used and the chromatograms were monitored at 285, 350 and 450 nm for the quantification 192 of phytoene, phytofluene and the rest of the carotenoids, i.e. lutein, lycopene and β193 16 Kasim & Kasim, 2015). In spite of its small size, ʽCherry cerejaʼ showed a similar TSC 334 and glucose content than the ʽTigerellaʼ, besides, ʽCherry amarilloʼ and ʽCherry pera 335 clásicoʼ has a similar glucose content than the ʽByelsaʼ This suggests that these cherry 336 varieties can become strong competitors for traditional varieties in terms of flavour. In 337 addition, an inverse correlation (with a value of -0.46 between TSC and weight) was 338 observed. These data keep relationship with other studies showing inverse correlations of 339 growth fruit rate and size with sugars (Coyago-Cruz, Corell, Moriana, et al., 2017). 340 3.2.2 Phenolics compounds 341 Data about total phenolic contents (TPC) and levels of individual compounds are 342 summarized in Table 1 and 2. TPC in cherry tomatoes ranged from 150.2 (ʽCherry cerejaʼ) 343 to 307.7 mg/100 g DW (ʽCherry pera naranjaʼ) (ca. 2-fold difference), while in common 344 tomatoes they ranged from 286.3 (ʽSunchocolaʼ) to 503.1 mg/100 g DW (ʽGreen Zebraʼ) 345 (ca. 1.8-fold difference). ʽCherry pera naranjaʼ and ʽGreen Zebraʼ were the varieties with 346 the highest TPC among cherry and tomato varieties, whilst ‘Cherry cereja’ and 347 ʽSunchocolaʼ, were those with the lowest levels, respectively. 348 TPC observed in red and yellow-orange cherry tomatoes ranged from 150.2 349 (ʽCherry cerejaʼ) to 239.8 (ʽCherry pera clásicoʼ) (ca. 1.6-fold difference) and from 263.5 350 (ʽCherry amarilloʼ) to 307.7 mg/100 g DW (ʽCherry pera naranjaʼ), respectively. These 351 values were similar or lower than those reported elsewhere (Cortés-Olmos, Leiva-Brondo, 352 Roselló, Raigónc, & Cebolla-Cornejo, 2014; Figàs et al., 2015). On the other hand, TPC in 353 dark tomatoes, i.e. ʽMinichocmato peraʼ (D) and ʽSunchocolaʼ (G), fluctuated between 354 220.9 and 286.3 mg/100 g DW. These values were similar or higher than those reported by 355 other authors (Choi et al., 2014; Cortés-Olmos et al., 2014) . 356 17 The values of TPC in red common tomatoes ranged from 292.6 (ʽPalamósʼ) to 344.9 357 mg/ 100 g DW (ʽTigerellaʼ) and they were in general similar or higher than those detected 358 in other similar tomato varieties (Cortés-Olmos et al., 2014; Periago, Martínez-Valverde, 359 Chesson, & Provan, 2002). On the other hand, ʽGreen Zebraʼ presented the highest value 360 of TPC within all the varieties under study and likewise greater values of p361 hydroxybenzoic, p-coumaric and chlorogenic acid. Finally, the common variety with 362 yellow colour showed markedly higher values of TPC (345.9 mg/100 g DW for ʽOrangeʼ), 363 relative to those found by other authors, who reported concentrations ranging from 57.2 to 364 251.2 mg of gallic acid equivalents/100 g DW in yellow and orange common tomatoes 365 (Cortés-Olmos et al., 2014) and lower than ranges reported by Raiola et al., i.e. 50.9 to 366 53.5 mg/100 g FW in yellow tomatoes. 367 Overall, there were not statistically significant differences in the values of TPC 368 between ʽTigerellaʼ and ʽOrangeʼ or between ʽPalamósʼ and ʽSunchocolaʼ in common 369 varieties. This fact indicates that common varieties other than red also provide significant 370 amounts of phenolic compounds, in addition these varieties showed higher contents than 371 the traditional varieties (ʽPalamósʼ) of between 1.2 and 1.7 times; this agreed with other 372 authors (Cortés-Olmos et al., 2014). Interestingly, an inverse correlation with a value of - 373 0.66 between size and TPC was observed. These data agree well with those reported in 374 other studies, who suggest that the size is inversely proportional with total flavonols 375 (Coyago-Cruz, Corell, Moriana, et al., 2017; Slimestada & Verheulb, 2009). On the other 376 hand, the TPC of ʽCherry pera naranjaʼ and ʽPalamósʼ were comparable, showing that 377 cherry varieties, despite their size, could be an important source of phenolic compounds. 378 In addition p-hydroxybenzoic acid, p-coumaric acid, caffeic acid, chlorogenic acid, 379 gallic acid, ferulic acid, naringin, crisin, quercetrin and quercetin were the major phenolic 380 18 compounds detected in the set of samples studied, which agreed well with the studies of 381 other authors (Periago et al., 2002; Raiola et al., 2015), while ferulic acid, naringin and 382 crisin were not found in cherry tomatoes while quercetrin was detected in traces . 383 Caffeic acid levels ranged from 3.9 (ʽMinichocmato peraʼ) to 20.7 mg/100 g DW 384 (ʽCherry amarilloʼ) (ca. 5-fold difference) and from 10.4 (ʽOrangeʼ) to 30.1 mg/100 g DW 385 (ʽSunchocolaʼ) (ca. 3-fold difference) in cherry and common varieties respectively. These 386 values were comparable to those found in other studies (Periago et al., 2002; Raiola et al., 387 2015).ʽCherry amarilloʼ and ʽSunchocolaʼ were the varieties with the highest contents of 388 this compound in cherry and common tomatoes, respectively. 389 Chlorogenic acid concentrations in cherries, ranged from 3.8 (ʽCherry cerejaʼ) to 390 68.5 mg/100 g DW (ʽCherry pera naranjaʼ) (ca. 18-fold difference) and were in general 391 lower than in common tomatoes (the levels in this group ranged from 6.4 (ʽSunchocolaʼ) to 392 84.9 mg/100 g DW (ʽGreen Zebraʼ) (ca. 13-fold difference)). Other authors have reported 393 concentrations of this compound between 1.4 and 236.0 mg/ 100 g FW in different 394 varieties of tomatoes (Periago et al., 2002; Raiola et al., 2015). 395 The quercetin concentrations fluctuated between 22.4 (ʽCherry cerejaʼ) and 49.6 396 mg/100 g DW (ʽCherry amarilloʼ) (ca. 2-fold difference) in cherry varieties, and between 397 25.8 (ʽOrangeʼ) and 62.1 mg/100 g DW (ʽTigerellaʼ) (ca. 2-fold difference) in common 398 tomatoes. Similar values were found by other authors (Choi et al., 2014; Periago et al., 399 2002; Raiola et al., 2015) . 400 3.2.3 Carotenoids 401 Quantitative data on individual and total carotenoids (TCC) are presented in Table 1 402 and 2. TCC observed in cherry tomatoes varied between 2.2 (‘Cherry amarillo’) and 102.0 403 19 mg/100 g DW (‘Minichocmato pera’) (ca. 50-fold difference), while in common tomatoes 404 they ranged from 11.8 (‘Sunchocola’) to 297.9 mg/100 g DW (‘Orange’) (ca. 30-fold 405 difference). Interestingly, the cherry varieties ʽMinichocmato peraʼ and ʽCherry cerejaʼ 406 showed higher TCC values than common varieties like ‘Green zebra’, ‘Sunchocola’ and 407 ‘Palamós’. Considering all the samples studied, the major carotenoids found were 408 phytoene, phytofluene, lutein, lycopene and β-carotene. Lycopene was the main carotenoid 409 in the varieties ʽMinichocmato peraʼ, ʽCherry cerejaʼ, ‘Tigerella’, ‘Byelsa’ and ‘Palamós’. 410 Phytoene was the predominant carotenoid in ʽCherry pera clásicoʼ, ‘Cherry pera naranja’, 411 ʽGreen Zebraʼ, ʽSunchocolaʼ and ʽOrangeʼ, whereas lutein was the most important 412 carotenoid in quantitative terms in ‘Cherry amarillo’. 413 The clear qualitative and quantitative differences observed not only in tomatoes but 414 also in other dietary fruits and vegetables are not surprising whatsoever as the levels of 415 secondary metabolites in general and carotenoids in particular are dependent on multiple 416 factors (genetic, climatic, agronomic, among others) (Dias et al., 2018). 417 The levels of the colourless carotenoid phytoene ranged from 0.3 (‘Cherry 418 amarillo’) to 252.6 mg/100 g DW (‘Orange’) (ca. 840-fold difference). These values were 419 comparable with the results presented by other authors, who found that common orange 420 varieties juice had higher phytoene contents than red varieties like TCC (Cooperstone et 421 al., 2015). Those of the colourless carotenoid phytofluene oscillated between non 422 detectable levels and 12.3 mg/100 g DW (‘Orange’). This latter carotenoid was not 423 predominant in any of the varieties surveyed. Tomatoes are indeed one of the best sources 424 of these largely ignored carotenoid rarities, which are attracting increasing interest due to 425 their likely health (protection against light-induced damage, anticarcinogenic activity, 426 20 protection against oxidation, among other) and cosmetic benefits (A.J. Meléndez-Martínez, 427 Mapelli-Brahm, & Stinco, 2018). 428 Lycopene was not detected in some of the varieties studied, whereas the highest 429 levels (117.1 mg/100 g DW) were found in the variety ʽTigerellaʼ. Tomatoes are usually 430 the main dietary source of this carotenoid that has been related to diverse health-promoting 431 actions (protection against light-induced damage, anticarcinogenic activity, protection in 432 cardiovascular disease, among others) in the last decades (Böhm, 2012; Giovannucci, 433 2002). On the other hand, the limitation of sucrose is thought to delay the accumulation of 434 lycopene and phytoene in the tomato pericarp (Li & Yuan, 2013) . The unavailability of 435 sucrose may explain the no detection of lycopene in ʽGreen Zebraʼ. Sucrose was not 436 detected in ʽSunchocolaʼ and ʽOrangeʼ either, varieties that contain lower lycopene levels 437 as compared to the other varieties of the common tomatoes. In ʽCherry amarilloʼ and 438 ʽCherry pera naranjaʼ, there was availability of sucrose but lycopene was not detected, This 439 might be due to the fact that the biosynthesis of carotenoids was beginning in these 440 varieties, which would suggest low degrees of ripening and would corroborate the initial 441 premise that these varieties were harvested without reaching physiological maturity. 442 The levels of the provitamin A carotenoid β-carotene ranged from 0.1 (‘Cherry 443 amarillo’) to 16.1 (ʽTigerellaʼ) (ca. 160-fold difference). This carotenoid was not 444 predominant in any of the varieties surveyed as the results of other authors show (Cortés445 Olmos et al., 2014). The higher content of β-carotene in ʽCherry amarilloʼ and ʽCherry 446 pera naranjaʼ, could be due to the fact that these varieties are thought to have not reached 447 physiological maturity. In this sense, it is to be considered that β-carotene is one of the 448 carotenoids present in photosynthetic tissues and therefore in stay-green tomatoes or those 449 that has not reached a high degree of ripening, which is typically accompanied by the large 450 21 accumulation of lycopene (Hernández-Gras, De-Pourcq, Angaman, & Boronat, 2017; 451 Antonio J. Meléndez-Martínez et al., 2010), and as also been shown in study in red cherry 452 varieties in different degrees of maturity, years, seasons and clusters (Coyago-Cruz et al., 453 2018). 454 3.3 Plastids morphology 455 The microscopic analysis revealed the existence of different types of plastids among 456 the samples. The most abundant substructures found in the different plastids were 457 plastoglobules and crystals remnants and the relative amount of them among varieties was 458 different. Several authors have suggested that plastoglobules in tomatoes are a source of 459 storage of β-carotene (Cooperstone et al., 2015; R. M. Schweiggert & Carle, 2017). 460 However other authors have suggested that β-carotene could also be present in crystalline 461 form (Harris & Spurr, 1969; Rosso, 1968; Ralf M. Schweiggert, Mezger, Schimpf, 462 Steingass, & Carle, 2012) mainly when there is a hyper-accumulation of this carotenoid in 463 the cells (Li & Yuan, 2013). On the other hand, lycopene is present in a solid crystalline 464 deposition form (Cooperstone et al., 2015; Hernández-Gras et al., 2017; Simkin et al., 465 2007), . In our study, this crystalline deposition form of lycopene was observed as 466 membranes with undulating shape in empty spaces, which are likely to be due to the 467 leaching out of the lycopene during the dehydration process (R. M. Schweiggert & Carle, 468 2017). The presence of plastoglobules in the varieties that contained no detectable amounts 469 of lycopene, i.e. ʽCherry amarilloʼ, ʽCherry pera naranjaʼ and ʽGreen Zebraʼ could be due 470 to the accumulation of β-carotene. On the other hand, the presence of crystals in ʽCherry 471 pera naranjaʼ could indicate that β-carotene was deposited in this form in this variety. 472 Chromoplasts in a relative early development stage were found in the greenish 473 common tomatoes, i.e. Green Zebra and Sunchocola varieties, and in the Byelsa variety 474 22 (Figures 3 -F, -G and –I), as these still contain chlorophyll pigments and therefore 475 chloroplasts (Hernández-Gras et al., 2017). Among other substructures, they contained 476 plastoglobules and crystal remnants. However, no crystal remnants were found in the 477 Green Zebra variety, which is likely to be due to the lack of lycopene (Table 2); the same 478 was observed in the micrographs corresponding to ʽCherry amarilloʼ (Figure 2-A and 2-C). 479 In these chromoplasts in a relative early development, starch granules, grana and 480 thylakoids with some degree of breakdown were also found. The presence of starch 481 granules in ʽByelsaʼ suggests that this variety has not yet reached full maturity, since 482 during tomato fruit ripening it has been demonstrated that there is a decline in starch 483 plastids and a progressive conversion into reducing sugars (Li & Yuan, 2013). On the other 484 hand, the absence of starch plastids in immature cherry varieties suggest that the presence 485 of carbohydrates was due to the degradation of starch and therefore the accumulation of 486 sugars was lower, as suggested by other authors (Beckles, 2012). 487 On the other hand, chloroplasts were found in the Minichocmato pera variety. In 488 these plastids the plastoglobules can be observed associated to the thylakoid membranes, 489 which has also been reported elsewhere (Li & Yuan, 2013; Shumskaya & Wurtzel, 2013) 490 (Figure 2-D). In the rest of the samples fully developed chromoplasts with different 491 carotenoid-accumulating structures were found. As can be observed in Figure 3-H, there 492 was a great accumulation of plastoglobules and crystal remnants in ʽTigerellaʼ, which 493 could be related with the fact that this variety was a richer source of lutein, β-carotene and 494 lycopene compared to the other varieties (Table 1 and 2). Peroxisomes containing 495 crystalline cores were noticed in ʽSunchocolaʼ, ʽTigerellaʼ ʽByelsaʼ and ʽPalamósʼ (Figures 496 3 -G, -H and -J). Peroxisomes are known to be multifaceted. Indeed they have been related 497 with processes such as photorespiration, nitrogen metabolism, detoxification, synthesis of 498 some plant hormones (Kaur et al., 2009) and modulation of molecular signals during fruit 499 23 ripening (Verlag et al., 2003). This might suggest that ʽTigerellaʼ, ʽByelsaʼ and ʽPalamósʼ 500 did not reach their maximum maturity and that the amount of lycopene could increase 501 since these varieties still have sucrose, which would favour biotransformation; however in 502 ʽSunchocolaʼ their presence might be related to some extent to detoxification, since this 503 variety was cultivated in autumn and the difficulty in cultivation due to the presence of 504 pests caused the application of chemicals that could cause plant poisoning.. In addition, in 505 the round red cherry several plastoglobules were found distributed along the membranes of 506 the chromoplasts (Figure 2-E). This could be related with the fact that carotenoids are 507 generated in the membrane of the plastids (Li & Yuan, 2013). All of the aforementioned 508 substructures were also found in other studies in different tomato varieties (Cooperstone et 509 al., 2015; Hernández-Gras et al., 2017; Simkin et al., 2007) and the differences found in 510 plastids among the different varieties of the same fruit could be due to some extent to 511 differences in the carotenoid profiles (R. M. Schweiggert & Carle, 2017), as, depending on 512 the carotenoid and its shape, the tendency for aggregation to eventually form crystals can 513 vary drastically. As an example, lycopene is a linear and rigid carotenoid with 11 c.d.b. 514 that is known to crystallize easily when is present in high amounts, even in organic 515 solvents, whereas the linear carotenes phytofluene and phytoene have fewer c.d.b. (5 and 516 3, respectively) and so they, have a less rigid shape and are not expected to crystallyze as 517 easily. On the other hand, as far as red tomatoes are concerned, lycopene occurs 518 predominantly as the (all-E)-isomer, which is more rigid and linear than the corresponding 519 Z isomers, whereas phytoene and phytofluene occur largely as Z isomers, hence the 520 tendency of the latter two tomato carotenes to aggregate and form crystals within the 521 chromoplast is even lower (Antonio J. Meléndez-Martínez, Paulino, Stinco, Mapelli522 Brahm, & Wang, 2014). 523 24 In addition, starch granules were clearly observed in ʽGreen Zebraʼ and ʽByelsaʼ 524 that exhibited a granular structure of the pulp. They were also present in ʽPalamósʼ, which 525 is a juicier variety. The presence of starch within the structure provides a certain thickening 526 character to the tomato pulp, which can interest for the pulp and sauces industry. In 527 relation to this, the declining of plastid starch content are correlated with fruit ripening 528 such that decreases in plastid starch are usually correlated with increases of carotenoids 529 and reducing sugars, which could explain the limited number of starch granules in the 530 varieties mentioned (Li & Yuan, 2013). In addition, an inverse relationship between the 531 content of lycopene and phytoene with the sucrose content was evidenced in this study 532 (Table 1 and 2), as also noted by other authors (Li & Yuan, 2013). 533 4. CONCLUSIONS 534 A comprehensive study of commercial quality parameters, sugars, phenols and 535 carotenoid accumulation in different tomato varieties have been carried out. The study of 536 the cherry and common varieties is particularly interesting due to the scarcity of studies in 537 varieties of tomato with coloration different from red. It has been concluded that, overall, 538 the commercial quality fruit parameter (weight and soluble solid) values in cherry varieties 539 were lower than the common varieties. 540 On the other hand, within the varieties studied ʽCherry cerejaʼ (524.1 mg/g DW), 541 ʽGreen Zebraʼ (523.9 mg/g DW) and ʽTigerellaʼ (522.7 mg/g DW) presented high values 542 of TSC. Besides, ʽCherry cerejaʼ showed a similar TSC and glucose content than 543 ʽTigerellaʼ. In addition, ʽCherry cerejaʼ showed high values of TSC associated mainly with 544 the accumulation of glucose, and ʽCherry amarilloʼ high values of fructose and sucrose. 545 The TPC values ranged from 150.2 (‘Cherry cereja’) to 503.1 mg/100 g DW (ʽGreen 546 Zebraʼ) (ca. 3.3-fold difference). p-Hydroxybenzoic acid, p-coumaric acid, caffeic acid, 547 25 chlorogenic acid, gallic acid, ferulic acid, naringin, crisin, quercetrin and quercetin were 548 the major phenolic compounds detected. The TCC ranged between 2.2 (‘Cherry amarillo’) 549 and 297.9 (‘Orange’) mg/100 g DW (ca. 150-fold difference). Lycopene was the major 550 carotenoid in ʽTigerellaʼ (117.1 mg/100 g DW). Phytoene was the predominant carotenoid 551 in ʽCherry pera clásicoʼ, ‘Cherry pera naranja’, ʽGreen Zebraʼ, ʽSunchocolaʼ and ʽOrangeʼ. 552 Plastids observation revealed the existence of different types of carotenoid-accumulating 553 substructures in the plastids among the samples. In general, the most abundant were 554 plastoglobules and crystals remnants, although the relative amount of them varied 555 considerably among varieties as a result of their colour and therefore of their carotenoid 556 profile. 557 AUTHOR INFORMATION 558 Corresponding Author 559 *Phone: +34-954557017 560 E-mail: [email protected] 561 562 Acknowledgements 563 The authors want to thank the Secretaría Nacional de Educación Superior, Ciencia, 564 Tecnología e Innovación (SENESCYT) - Ecuador for its financial support and the 565 Technical Staff of the Service of Biology (SGI, Universidad de Sevilla). AJMM 566 acknowledges funding from the Spanish State Secretariat of Research, Development and 567 Innovation (Ministry of Economy and Competitiveness, project ref. AGL2012-37610, co568 funded by FEDER). ECC, DH, CMS and AJMM thank the Ibero-American Programme for 569 Science, Technology and Development (CYTED, http://www.cyted.org) for the funding of 570 Fig. 3 . Imágenes de microscopía electrónica de plástidos y otras estructuras en tomates comunes con diversos colores. La barra en cada figura representa la escala de tamaño para esa figura. 'Green Zebra' (F), 'Sunchocola' (G), 'Tigerella' (H), 'Byelsa' (I), 'Palamós' (J), 'Orange' (K). 1, membrana externa ; 2, membrana interna ; 3, gránulos de almidón; 4, los restos de cristal; 5, grana; 6, gotas lipídicas; 7, membranas tilacoides ; 8, plastoglobules; MT, mitocondrias ; P, peroxisoma; CW, pared celular. Nota: La variedad naranja (K) no aparece en la figura, ya que no estaba disponible en el momento de los análisis microscópicos. (Para la interpretación de las referencias al color en la leyenda de esta figura, se remite al lector a la versión web de este artículo). Tabla 1 . Valores promedio de parámetros de calidad comercial, azúcares, fenólicos y carotenoides de tomates cherry . 'Cherry Amarillo' (A) 'Cherry pera clásico' (B) 'Cereza Pera Naranja' (C) 'Minichocmato pera' (D) 'Cherry cereja' (E) Ac Color Amarillo rojo naranja Verde rojo rojo Parámetros de calidad ED (cm) 4.2 ± 0.5 b 3.6 ± 0.6 d 3.7 ± 0.8 d 3.7 ± 0.4 cd 4.3 ± 0.2 a *** LD (cm) 2.6 ± 0.2 d 3.6 ± 0.3 a 3.4 ± 0.2 b 3.0 ± 0.4 c 2.9 ± 0.2 c *** Peso (gramos) 9.6 ± 1.9 b 9.0 ± 1.9 b 11.3 ± 3.7 b 7.9 ± 1.6 c 11.5 ± 1.0 a *** SS (° Brix) 3.5 ± 1.0 ab 3.3 ± 0.8 ab 3.3 ± 0.8 ab 3.3 ± 1.0 ab 3.7 ± 0.5 a * L * 44.6 ± 0.8 b 34.0 ± 1.9 d 49.1 ± 0.9 a 31.5 ± 1.4 e 35.6 ± 1.7 c *** C * ab 40.9 ± 4.0 b 38.0 ± 2.6 c 53.0 ± 2.9 a 18.3 ± 2.5 d 41.3 ± 4.0 b *** h ab 81.2 ± 3.1 a 40.9 ± 3.0 d 68.8 ± 0.8 b 66.2 ± 10.4 c 41.7 ± 2.5 d *** Carotenoides (mg / 100 g DW) y Fitoeno 0.3 ± 0.1 d 8.1 ± 0.0 c 25.4 ± 0.1 a 11.6 ± 1.2 b 14.1 ± 0.0 b ** Fitoflueno Dakota del Norte 0.7 ± 0.0 b 3.2 ± 0.2 a Dakota del Norte 3.2 ± 0.0 a Luteína 1.0 ± 0.0 c 0.5 ± 0.0 d 0.6 ± 0.0 d 6.1 ± 0.1 a 1.6 ± 0.0 b *** Licopeno Dakota del Norte 4.7 ± 0.1 c Dakota del Norte 77.5 ± 3.3 a 69.2 ± 0.9 b β-caroteno 1.2 ± 0.0 c 0.5 ± 0.0 d 0.5 ± 0.0 c 6.9 ± 0.2 a 2.4 ± 0.0 b *** TCC 2.5 ± 0.1 d 14.4 ± 0.1 c 29.7 ± 0.2 b 102.0 ± 3.5 a 90.7 ± 0.9 a *** Compuestos fenólicos (mg / 100 g DW) y p -hidroxi 130.2 ± 1.19 d 173.5 ± 0.1 a 165.0 ± 1.8 b 136.4 ± 0.9 c 86.2 ± 0.7 e *** p -Cumar 30.7 ± 1.1 b 6.3 ± 0.1 c 6.4 ± 0.0 c 33.2 ± 0.2 a 5,6 ± 0,2 c *** Cafeína 20.7 ± 1.5 a 4.3 ± 0.0 c 15.3 ± 0.8 b 3.9 ± 0.1 c 20.3 ± 0.2 a *** Chloroge 23.1 ± 1.7 b 4.3 ± 0.1 d 68.5 ± 1.5 a 13.3 ± 0.1 c 3.8 ± 0.2 d *** gálico 9.3 ± 0.1 c 12.8 ± 0.4 a 10.6 ± 0.4 b 7.8 ± 0.0 d 12.0 ± 0.1 a *** Ferulico Dakota del Norte Dakota del Norte Dakota del Norte Dakota del Norte Dakota del Norte Naringin Dakota del Norte Dakota del Norte Dakota del Norte Dakota del Norte Dakota del Norte Crisina Dakota del Norte Dakota del Norte Dakota del Norte Dakota del Norte Dakota del Norte Quercetrin tr tr tr tr tr Quercetina 49.6 ± 0.7 a 38.4 ± 1.2 c 42.0 ± 0.7 b 26.4 ± 0.6 d 22.4 ± 0.7 e *** TPC 263.5 ± 0.8 b 239.8 ± 1.8 c 307.7 ± 0.1 a 220.9 ± 0.2 d 150.2 ± 2.0 e *** Azúcares z (mg / g DW) y Fructosa 114.1 ± 1.7 a 91.2 ± 2.7 b 95.5 ± 0.4 b 58.8 ± 0.9 d 74.1 ± 1.4 c *** Glucosa 320.4 ± 8.0 c 315.9 ± 11.3 c 359.9 ± 0.1 b 214.9 ± 1.8 d 426.0 ± 1.3 a *** Sacarosa 55.1 ± 1.3 a 30.9 ± 0.0 b 30.5 ± 0.1 b 34.6 ± 3.3 b 23.9 ± 0.5 c *** TSC 489.6 ± 11.0 b 438.0 ± 14.0 c 485.9 ± 0.3 b 308.4 ± 6.0 d 524.1 ± 0.4 a *** Valores medios ± SD; [ x (n = 40); y (n = 12)]. La importancia de las diferencias entre las variedades de cereza (A C ), se da: ns, no significativo; * , p <0.1; **, p <0,01; ***, p <0,001. Los valores medios seguidos por la misma letra no difieren significativamente en el nivel de confianza del 99% dado. tr, traza; nd, no detectable; DE, diámetro ecuatorial; LD, diámetro longitudinal; SS, sólido soluble; TCC, carotenoides totales; pHidroxi, ácido p-hidroxibenzoico; p -Cumar, p - ácido cumárico; Cafeico, acido cafeico ; Chloroge, ácido clorogénico; Ácido gálico , gálico ; Ferulic, ácido ferúlico ; TPC,compuestos fenólicostotales; CET, contenido total de azúcares. Tabla 2 . Valores medios de parámetros de calidad comercial, azúcares, fenólicos y carotenoides de tomates comunes. 'Cebra Verde' (F) 'Sunchocola' (G) 'Tigerella' (H) 'Byelsa' (I) 'Palamós' (J) 'Naranja' (K) AH Unch Color Verde amarillo Verde rojo Rojo-amarillo rojo rojo naranja Parámetros de calidad ED (cm) 9.2 ± 1.7 b 4.6 ± 0.2 d 7.2 ± 0.6 c 6.8 ± 0.7 c 9.6 ± 0.7 b 13.7 ± 2.2 a *** *** LD (cm) 5.0 ± 0.7 c 3.9 ± 0.1 d 4.0 ± 0.4 d 6.2 ± 1.1 b 5.1 ± 0.3 c 7.0 ± 0.6 a *** *** Peso (gramos) 94.7 ± 40.3 b 50.4 ± 6.9 c 45.6 ± 6.2 c 56.7 ± 15.7 c 102.8 ± 19.4 b 274.9 ± 12.6a *** *** SS (° Brix) 6.2 ± 0.5 a 5.6 ± 0.9 a 4.6 ± 0.9 b 5.7 ± 1.0 a 4.6 ± 0.5 b 6.0 ± 0.5 a *** *** L * 44.4 ± 6.1 b 34.1 ± 0.9 e 36.7 ± 3.3 d 39.2 ± 4.2 c 43.3 ± 3.3 b 50.7 ± 4.7 a *** *** C * ab 41.9 ± 8.5 b 17.0 ± 1.7 d 35.4 ± 4.7 c 44.8 ± 5.0 b 44.2 ± 8.8 b 62.3 ± 2.8 a *** *** h ab 96.0 ± 3.0 a 58.7 ± 3.8 c 46.0 ± 3.0 e 44.1 ± 6.8 e 52.6 ± 5.8 d 62.7 ± 6.2 b *** *** Carotenoides (mg / 100 g DW) y Fitoeno 45.9 ± 8.1 b 12.1 ± 0.4 d 27.2 ± 4.5 c 23.5 ± 4.7 c 21.7 ± 4.0 c 252.6 ± 21.2a *** *** Fitoflueno Dakota del Norte Dakota del Norte 2.3 ± 0.4 b rastro traza ± 12.3 ± 0.7 a ** *** Luteína 0.8 ± 0.1 d 3.8 ± 0.8 a 3.6 ± 0.4 a 2.8 ± 0.5 b 1.6 ± 0.2 c 0.4 ± 0.1 e *** *** Licopeno Dakota del Norte 15.0 ± 0.0 d 117.1 ± 23.7a 110.4 ± 11.2a 47.4 ± 8.0 b 30.5 ± 0.6 c *** *** β-caroteno 1.1 ± 0.1 e 4.5 ± 0.1 d 16.1 ± 1.0 a 7.0 ± 0.6 c 10.9 ± 0.3 b 1.9 ± 0.2 e *** *** TCC 50.2 ± 8.4 d 35.5 ± 0.3 e 166.4 ± 2.6b 143.7 ± 15.0b 80.9 ± 9.2 c 297.9 ± 20.7a *** *** Compuestos fenólicos (mg / 100 g DW) y p -hidroxi 183.9 ± 9.7 a 102.9 ± 8.4 c 83.7 ± 1.0 d 69.7 ± 1.6 e 68.2 ± 1.1 e 147.2 ± 9.2 b *** *** p -Cumar 104.0 ± 11.3a 31.8 ± 3.7 c 31.6 ± 0.5 c 19.7 ± 1.7 d 24.0 ± 1.1 c 58.5 ± 2.5 b *** *** Cafeína 13.9 ± 0.7 bc 30.1 ± 2.2 a 17.0 ± 0.3 b 11.0 ± 0.3 c 10.6 ± 0.5 c 10.4 ± 1.4 c *** *** Chloroge 85.0 ± 1.1 a 6.4 ± 0.1 e 68.9 ± 5.0 b 74.5 ± 3.2 ab 64.8 ± 8.3 c 40.9 ± 2.2 d *** *** gálico 14.8 ± 1.2 b 21.7 ± 0.5 a Dakota del Norte Dakota del Norte Dakota del Norte 9.3 ± 0.2 c *** *** Ferulico 15.5 ± 2.4 a Dakota del Norte 12.8 ± 0.9 b 11.5 ± 1.8 bc 10.1 ± 0.5 c 9.2 ± 0.3 d *** *** Naringin 9.5 ± 1.0 b 2.6 ± 0.0 d 9.5 ± 0.4 c 24.7 ± 6.2 a 13.9 ± 0.7 b Dakota del Norte *** *** Crisina 31.8 ± 0.6 b 37.5 ± 0.7 a 31.6 ± 0.3 b 32.7 ± 0.8 b 32.6 ± 0.8 b 32.1 ± 0.3 b *** *** Quercetrin 18.1 ± 0.5 c 15.6 ± 0.3 cd 27.7 ± 1.1 ab 29.5 ± 0.7 a 26.1 ± 1.9 b 14.9 ± 1.1 d *** *** Quercetina 30.4 ± 0.6 c 37.9 ± 0.4 bc 62.1 ± 2.6 a 61.2 ± 8.2 a 42.2 ± 2.7 b 25.8 ± 1.8 d *** *** TPC 503.1 ± 5.7 a 286.3 ± 3.5 d 344.9 ± 7.6b 334.5 ± 12.4c 292.6 ± 12.9 d 345.9 ± 19.2b *** *** Azúcares z (mg / g DW) y Fructosa 132.7 ± 2.8 b 94.5 ± 2.2 c 83.1 ± 0.3 d 99.1 ± 3.6 c 81.3 ± 1.3 d 156.6 ± 0.7 a *** *** Glucosa 391.2 ± 5.0 b 288.1 ± 8.5 c 416.0 ± 6.5a 313.2 ± 14.6c 406.8 ± 1.2 a 307.5 ± 1.7 c *** *** Sacarosa Dakota del Norte Dakota del Norte 23.6 ± 0.7 b 30.0 ± 0.5 a 23.0 ± 0.4 b Dakota del Norte ns *** TSC 523.9 ± 7.8 a 410.2 ± 12.6d 522.7 ± 7.0ab 442.4 ± 18.6cd 511.2 ± 0.4 b 464.1 ± 1.8 c *** *** Valores medios ± SD; [ x (n = 21); y (n = 12)]. La importancia de las diferencias entre los tomates comunes (A H ) y todas las variedades (A CH) se da: ns, no significativo; *, p <0.1; **, p <0,01; ***, p <0,001. Los valores medios seguidos por la misma letra no difieren significativamente en el nivel de confianza del 99% dado. tr, traza; nd, no detectable; DE, diámetro ecuatorial; LD, diámetro longitudinal; SS, sólido soluble; TCC, carotenoides totales; p-Hidroxi, ácido phidroxibenzoico; p -Cumar, p -ácido cumárico; Cafeico, acido cafeico ; Chlorogeácido clorogénico ; Ácido gálico , gálico ; Ferulic, ácido ferúlico ; TPC,compuestos fenólicostotales; CET, contenido total de azúcares.