Temporal sequence of cell wall disassembly events in developing fruits. 2. Analysis of blueberry (Vaccinium species)

Softening and pathogen susceptibility are the major factors limiting the marketing of blueberries as fresh fruits, and these traits are associated with fruit cell wall structure. However, few studies that characterize wall modifications occurring during development and ripening have been reported for this fruit. In this study the ripening-associated modifications of blueberry fruit cell walls (composition, pectin and hemicellulose solubilization, and depolymerization) at five stages of ripeness have been analyzed. Xylose was found to be the most abundant noncellulosic neutral sugar associated with fruit walls, and the observed high Xyl/Glc ratio suggested that xylans, which are usually a minor hemicellulosic fruit wall component, are abundant in blueberry. The pectic matrix showed increased solubilization at early and intermediate stages of ripening, but no changes were detected in late ripening. Furthermore, little reduction in pectin polymer size occurred during blueberry ripening. In contrast, hemicellulose levels decreased as ripening progressed, and a clear depolymerization of these components was observed. A model for cell wall degradation in this fruit is discussed.     

Compositional changes in 'Bartlett' pear ( Pyrus communis L.) cell wall polysaccharides as affected by sunlight conditions

Preharvest conditions can have a great impact on fruit quality attributes and postharvest responses. Firmness is an important quality attribute in pear, and excessive softening increases susceptibility to bruising and decay, thus limiting fruit postharvest life. Textural characteristics of fruits are determined at least in part by cell wall structure and disassembly. Few studies have analyzed the influence of fruit preharvest environment in softening, cell wall composition, and degradation. In the current work 'Bartlett' pears grown either facing the sun (S) or in the shade (H) were harvested and stored for 13 days at 20 °C. An evaluation of fruit soluble solids, acidity, color, starch degradation, firmness, cell wall yield, pectin and matrix glycan solubilization, depolymerization, and monosaccharide composition was carried out. Sun-exposed pears showed more advanced color development and similar levels of starch degradation, sugars, and acids than shaded fruit. Sunlight-grown pears were at harvest firmer than shade-grown pears. Both fruit groups softened during storage at 20 °C, but even after ripening, sun-exposed pears remained firmer. Sunlight exposure did not have a great impact on pectin molecular weight. Instead, at harvest a higher proportion of water-solubilized uronic acids and alkali-solubilized neutral sugars and a larger mean molecular size of tightly bound glycans was found in sun-exposed pears. During ripening cell wall catabolism took place in both sun- and shade-grown pears, but pectin solubilization was clearly delayed in sun-exposed fruit. This was associated with decreased removal of RG I-arabinan side chains rather than with reduced depolymerization.     

Changes in sugars,acids, and volatiles during ripening of koubo [Cereus peruvianus (L.) Miller] fruits

The columnar cactus Cereus peruvianus (L.) Miller, Cactaceae (koubo), is grown commercially in Israel. The unripe fruits are green, and the color changes to violet and then to red when the fruit is fully ripe. The content of soluble sugars was found to increase 5-fold during ripening. Glucose and fructose were the main sugars accumulated in the fruit pulp, and each increased from 0.5 to 5.5 g/100 g fresh weight during ripening. The polysaccharides content decreased during ripening from 1.4 to 0.4 g/100 g fresh weight. The titratable acidity decreased and the pH increased during ripening. The major organic acid found in the fruit was malic acid, which decreased from 0.75 g/100 g fresh weight at the mature green stage to 0.355 g/100 g fresh weight in ripe fruits. Citric, succinic, and oxalic acids were found in concentrations lower than 0.07 g/100 g fresh weight. Prominent accumulation of aroma volatiles occurred toward the end of the ripening process. The main volatile found in the ripe fruit was linalool, reaching concentrations of 1.5-3.5 microg/g fresh weight.     

Tissue-specific and developmental modifications of grape cell walls influence the adsorption of proanthocyanidins

Cell wall material from Vitis vinifera L. cv. Cabernet Sauvignon grape skin and flesh was isolated at different stages of grape maturity to determine whether developmental changes in cell wall composition in different tissue types influence the binding of proanthocyanidins (PAs). Trends in cell wall adsorption of, and selectivity for, PAs were determined using two skin PAs that differed in their average molecular masses. Flesh cell walls consistently bound a higher amount of PA than those from skin. Key structural differences that reduced PA adsorption in skin cell walls by comparison with flesh cell walls were endogenously higher concentrations of insoluble PA, Klason lignin, and lower cell wall-bound protein. These differences may confer reduced flexibility and porosity of skin cell walls relative to flesh cell walls. Analysis of skin and flesh cell wall properties revealed that the onset of ripening was associated with a loss of type I arabinogalactan and galacturonic acid, which indicated a degradation of pectin within the cell wall. Flesh cell walls consistently bound PAs of larger molecular mass, and changes in PA adsorption properties after the onset of ripening were minor. For skin cell walls, adsorption of PA was lowest immediately following solubilization of galacturonic acid, and high molecular mass PAs were poorly bound. As ripening progressed, PAs of higher molecular mass were selectively adsorbed by skin cell walls, which indicates that ongoing cell wall remodeling during ripening may confer an increased porosity within the skin cell wall matrix, resulting in a greater adsorption of PA within a permeable structure.     

Simultaneous transgenic suppression of LePG and LeExp1 influences fruit texture and juice viscosity in a fresh market tomato variety

Tomatoes are grown for fresh consumption or for processing of the fruit. Some ripening-associated processes of the fruit can either contribute to or degrade attributes associated with both fresh and processing quality. For example, cell wall disassembly is associated with loss of fresh fruit firmness as well as with loss of processed tomato product viscosity. Several enzymes contribute to cell wall polysaccharide disassembly. Polygalacturonase (PG, poly[1,4-alpha-d-galactouronide] glucanohydrolase, EC 3.2.1.15) is among the most abundant polysaccharide hydrolases in ripening tomato fruit and is the major contributor to pectin depolymerization. Expansin (LeExp1) is also abundant in ripening fruit and is proposed to contribute to cell wall disassembly by nonhydrolytic activity, possibly by increasing substrate accessibility to other enzymes. Suppression of either LePG or LeExp1 expression alone results in altered softening and/or shelf life characteristics. To test whether simultaneous suppression of both LePG and LeExp1 expression influences fruit texture in additive or synergistic ways, transgenic Lycopersicon esculentum var. Ailsa Craig lines with reduced expression of either LePG or LeExp1 were crossed. Fruits from the third generation of progeny, homozygous for both transgenic constructs, were analyzed for firmness and other quality traits during ripening on or off the vine. In field-grown transgenic tomato fruit, suppression of LeExp1 or LePG alone did not significantly increase fruit firmness. However, fruits suppressed for both LePG and LeExp1 expression were significantly firmer throughout ripening and were less susceptible to deterioration during long-term storage. Juice prepared from the transgenic tomato fruit with reduced LePG and LeExp1 expression was more viscous than juice prepared from control fruit.     

Impact of the stage of ripening and dietary fat on in vitro bioaccessibility of beta-carotene in 'Ataulfo' mango

Pulp from "slightly ripe", "moderately ripe", or "fully ripe" mangoes was digested in vitro in the absence and presence of processed chicken as a source of exogenous fat and protein to examine the impact of stage of ripening of mango on micellarization during digestion and intestinal cell uptake (i.e., bioaccessibility) of beta-carotene. The quantity of beta-carotene transferred to the micelle fraction during simulated digestion significantly increased as the fruit ripened and when chicken was mixed with mango before digestion. Qualitative and quantitative changes that occur in pectin from mango pulp during the ripening process influenced the efficiency of micellarization of beta-carotene. Finally, the uptake of beta-carotene in micelles generated during simulated digestion by Caco-2 human intestinal cells confirmed the bioaccessibility of the provitamin A carotenoid in mango.     

Impact of UV-C irradiation on the cell wall-degrading enzymes during ripening of tomato (Lycopersicon esculentum L.) fruit

The effect of a hormic dose of UV-C (254 nm) on changes in fruit firmness and cell wall-degrading enzyme (CWDE) activity was determined using tomato fruit. Throughout the storage period, a decrease in firmness was jointly observed with an increase of the CWDE (polygalacturonase, pectin methyl esterase, cellulase, xylanase, beta-D-galactosidase, and protease) activity for all treatments, suggesting the involvement of these enzymes in the ripening process. However, the enhancement in the activity of the CWDE was significantly less in fruit subjected to the hormic dose of UV-C. This reduction may explain why irradiated fruit were firmer than control and consequently may explain how UV-C could delay the ripening and senescence process. We suggest that the CWDE are one of the targets of the UV-C, and by this action, irradiation contributed to a delay of the cell wall degradation and consequently retarded softening of the tomato fruit tissues.     

猕猴桃果实贮藏期间细胞壁多糖物质降解特性及组织结构差异分析

为了深入了解猕猴桃果实采后细胞壁多糖物质降解及组织结构变化与果实贮藏性的关系,本研究以红阳和华特猕猴桃果实为试材,对25和4℃贮藏期间的细胞壁多糖物质含量及果胶降解酶活性进行测定,并比较两品种果实在25℃贮藏期间的细胞显微结构和钙组分含量差异。结果表明,在25和4℃贮藏条件下,随着贮藏时间的延长,红阳和华特猕猴桃果实中半纤维素（HCL）、纤维素（CL）和共价结合型果胶（CSP）含量不断降低,水溶性果胶（WSP）含量不断上升,离子结合型果胶（ISP）含量相对稳定。红阳猕猴桃各细胞壁多糖组分含量变化速度较快。两品种猕猴桃果实硬度均与WSP含量呈显著负相关,与CSP含量呈显著正相关。果胶降解酶活性检测结果显示,25℃贮藏前期,红阳猕猴桃中果胶酸裂解酶（PL）和β-半乳糖苷酶（β-Gal）活性显著高于华特;贮藏中后期,红阳中果胶甲酯酶（PME）活性显著高于华特。4℃贮藏期间,红阳中PME活性仍显著高于华特;4℃贮藏前期,红阳中β-Gal活性与华特无显著差异,而PL活性低于华特。相关性分析表明,25℃贮藏期间,与红阳和华特果实软化显著相关的果胶降解酶分别是PME和PL;4℃贮藏期间,与华特果实软...     

钙处理对葡萄柚果实细胞壁物质代谢及其相关基因表达的影响

【目的】研究采前、 采后钙处理对葡萄柚果实细胞壁组分、 细胞壁降解酶活性变化及其相关基因表达的影响,可为了解钙与果实细胞壁物质代谢之间的关系,揭示钙对果实软化的作用机理,为调控葡萄柚果实膳食纤维含量,提高果实质地品质提供理论依据。【方法】试验于2011年2月至11月在云南省玉溪市葡萄柚果园进行,供试品种为‘里约红’葡萄柚,该品种于2005年嫁接于当地砧木,株行距为3 m×3 m。试验由采前和采后钙处理两部分组成。采前钙处理在幼果初期、 幼果末期、 膨大初期、 膨大末期、 转色期,叶面喷施2% CaCl2; 采后钙处理在果实成熟采后浸于2% CaCl2溶液5 min, 室温贮藏。之后每15天取样一次,每次取10个果实,测定葡萄柚果肉细胞壁组分、 细胞壁降解酶活性及其基因表达量。【结果】随葡萄柚果实后熟软化,紧密结合型果胶(共价结合型果胶)解聚为松散结合型果胶(水溶性果胶、 离子结合型果胶),紧密结合型半纤维素(24% KOH可溶性半纤维素)解聚使其含量下降,而松散结合型半纤维素含量增加(4%KOH可溶性半纤维素)。果实PG、 PME、 Cx、 α-L-Af和β-Gal酶活性及其基因表达量均随果实软化呈不同程度增加。PME活性在果实采收后表现出较高含量,而PG活性在果实贮藏前期急剧增加,其酶基因的表达量与酶活性变化趋势基本一致。Cx、 α-L-Af和β-Gal活性在贮藏中、 后期上升较快,相关酶基因的表达量亦明显增加。钙处理显著地降低果实细胞壁降解酶活性和基因表达水平,其中采后钙处理对α-L-Af和β-Gal活性和基因表达在贮藏中、 后期的调控作用较显著,酶活性和基因表达均维持在较低水平。【结论】外源钙处理降低细胞壁降解酶活性及其基因表达,抑制了细胞壁物质的解聚,采后钙处理对细胞壁物质代谢的调控效果优于采前钙处理。外源钙处理抑制了细胞壁降解酶基因表达水平,降低了细胞壁降解酶活性,减缓了果胶、 半纤维素的解聚,从而达到调控果实膳食纤维含量、 维持果实质地品质、 延长果实货架期寿命的目的。     

Metabolism of phenolic compounds during loquat fruit development.

Phenolic compounds in loquat fruit were identified as 5-caffeoylquinic acid (chlorogenic acid), neochlorogenic acid, hydroxybenzoic acid, 5-p-feruloylquinic acid, protocatechuic acid, 4-caffeoylquinic acid, epicatechin, o-coumaric acid, ferulic acid, and p-coumaric acid. Neochlorogenic acid was found to be dominant in the early stages of loquat fruit development. Both the concentrations and types of phenolic compounds were high in young fruit but then decreased steadily during growth. However, the concentration of chlorogenic acid increased during ripening and became predominant in ripe fruit. The large rise in chlorogenic acid concentration appears to be a characteristic of loquat fruit ripening. In all of the cultivars tested, the types of phenolic compounds were similar but the total phenolic content varied from 81.8 to 173.8 mg/100 g of fresh pulp. In the biosynthetic pathway of chlorogenic acid, the enzyme activities of phenylalanine ammonia-lyase (PAL), 4-coumarate:CoA ligase (CL), and hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase (CQT) were high at the early stage of growth, diminished to low levels approximately 3 weeks prior to harvest, but then rose to a peak at 1 week before harvest. The changes of these enzyme activities seemed to be associated with variations in chlorogenic acid concentration during development, maturation, and ripening of loquat fruit.     

Antioxidant systems and their relationship with the response of pepper fruits to storage at 20 degrees C

Fresh peppers (Capsicum annuum L., variety California) in their green and red ripe stages were stored at 20 degrees C for 7 and 19 days to determine the effects of storage on whole fruit antioxidant capacity (TAA) and ascorbate (ASC) content, as well as on some antioxidant enzyme activities, such as catalase (CAT), superoxide dismutase (SOD), and those of the ASC-glutathione cycle. At least one Mn-SOD, two Fe-SODs, and three CuZn-SODs were detected in the fruit extract after native polyacrylamide gel electrophoresis. All of the SOD isozymes and glutathione reductase had higher activity levels in the red control fruits than in the green fruits, whereas the activities of monodehydroascorbate and dehydroascorbate reductase were higher in green fruits. Ascorbate peroxidase (APX) was found to be similar in both fruits. SODs, CAT, and APX seem to be involved in pepper fruit ripening and senescence during storage at 20 degrees C, perhaps influencing the active oxygen species levels in the fruit. TAA, as well as the ASC content, was higher in red peppers than in green, and storage increased the ASC in both green and red fruits.     

Effect of dips in a 1-methylcyclopropene-generating solution on 'Harrow Sun' plums stored under different temperature regimes

The effect of postharvest dips in a 1-methylcyclopropene-generating solution of the formulation AFxRD-038 (Rohm & Haas) on plum fruit (Prunus salicina Lindell cv. 'Harrow Sun') quality and ripening during storage was determined. Fruit weight loss, tissue firmness, soluble solids content (SSC), titratable acidity (TA), ethylene production, respiration, and the activities of the cell wall modifying enzymes polygalacturonase (PG), 1,4-beta-D-glucanase/glucosidase (EGase), beta-galactosidase (beta-gal), and pectin methylesterase (PME) were measured. Fruit reddening, anthocyanin content, and phenylalanine ammonia-lyase (PAL) activity were also analyzed. The 1-MCP-treated fruit showed reduced ethylene production and respiration rate and delayed softening, which was associated with the reduction in the activity of PG, EGase, and beta-gal. The immersion in 1-MCP-generating solutions also decreased weight and acidity loss without modifying the fruit SSC. The immersion treatment was particularly effective in the fruit stored at 5 degrees C, keeping higher overall quality, maintaining lower levels of anthocyanins and PAL activity, and preventing flesh reddening. The present data show that beneficial effects in delaying plum fruit ripening and controlling chilling injury can be obtained by dipping the fruits in a solution of this novel 1-MCP-generating formulation.     

Olive fruit cell wall: degradation of pectic polysaccharides during ripening

Olive fruits at three stages of ripening (green, cherry, and black) have been studied. After cell wall isolation, the compositions of the cell wall and that of the phosphate-soluble polysaccharides were determined. In cell walls, decreases in arabinose, xylose, glucose, and uronic acid levels were observed, together with a slight increase in mannose on ripening. At the beginning of ripening, fragments of pectic polymers were the major constituents of the phosphate-soluble fraction, with the hemicellulosic ones increasing toward the end of the process. The molecular weight of the fragments solubilized was approximately 6 kDa. After cell wall fractionation, the pectic polysaccharides soluble in imidazole and sodium carbonate were also studied. In both fractions, between the green and cherry stages of ripening, a significant loss of homogalacturonans took place. Between the cherry and black stages of ripening, rhamnogalacturonan side chains were also released in addition to homogalacturonans. In any of the pectic fractions, changes in apparent molecular weight were quantified.     

Olive fruit cell wall: degradation of cellulosic and hemicellulosic polysaccharides during ripening

Cellulose and hemicelluloses obtained from the cell walls of partially depectinated olives have been studied at three stages of ripening (green, cherry, and black). Hemicelluloses were fractionated into two groups, the amounts of which diminished during ripening: those soluble in 4% KOH diminished between the cherry and black stages, whereas those soluble in 24% KOH did so between the green and cherry stages. Arabinoxylans, xyloglucans, and homo- and/or rhamnogalacturonans to a lesser extent were present in these fractions. After ion exchange and size exclusion chromatographies, decreases in the molecular weights of hemicelluloses, mainly in the neutral fractions, were observed. The amount of cellulose also decreased, but at the second stage of the ripening process. Approximately 2 mg/fruit of glucose was lost from cellulose, and the amount of uronic acids increased (0.23 mg/fruit).     

Identification of new strawberry sulfur volatiles and changes during maturation

Two Florida strawberry cultivars, 'Strawberry Festival' and 'Florida Radiance', were harvested at five fruit developmental stages (white, half red, three-quarter red, full ripe, and overripe) at four harvest dates. A static headspace solid-phase microextraction (SPME) sampling technique coupled with gas chromatography (GC) using pulsed flame photometric detection (PFPD) was employed to measure 16 sulfur volatiles in these strawberries. A total of 7 sulfur volatiles have been previously reported, and 9 are reported for the first time in strawberries. Newly identified sulfur volatiles include methyl thiopropionate, ethyl thiobutanoate, methyl thiohexanoate, methyl (methylthio)acetate, ethyl (methylthio)acetate, methyl 2-(methylthio)butyrate, methyl 3-(methylthio)propionate, ethyl 3-(methylthio)propionate, and methyl thiooctanoate. Identifications were based on matching sulfur peak linear retention indexes (LRIs) of unknowns with authentic standards and gas chromatography-mass spectrometry (GC-MS) data. Concentrations were determined using both internal and external standards. Most sulfur volatiles increased with increasing maturity, with only concentrations of hydrogen sulfide and methanethiol remaining relatively consistent at all five stages. At the white and half red stages, most sulfur volatiles consisted of various alkyl sulfides. At three-quarter red (commercial ripe), full ripe, and overripe stages, the majority of sulfur volatiles consisted of sulfur esters. Most sulfur volatiles increased dramatically between the commercial ripe, full ripe, and overripe stages, increasing as much as 100% between full ripe and overripe. Principal component analysis indicated that sulfur volatiles could be used to distinguish overripe from full ripe and commercial ripe berries.     

Determination and isolation of a thioesterase from passion fruit (Passiflora edulis Sims) that hydrolyzes volatile thioesters

Volatile organosulfur compounds (VOSCs) are high impact aroma chemicals characteristic of tropical fruits which are active as both free thiols and the respective thioesters. Using a simple and sensitive colorimetric enzyme assay, a thioesterase activity toward VOSCs has been identified in ripening purple passion fruit ( Passiflora edulis Sims). The assay was based on determining the release of free thiols from 2-methyl-3-furanthiol acetate using Ellman's reagent. The major thioesterase in the fruit was found to be a wall-bound protein in the mesocarp. The extracted enzyme activity was purified 150-fold and shown to be associated with a 43 kDa monomeric serine hydrolase which was selectively labeled with a fluorophosphonate suicide probe. MS-MS sequencing identified the thioesterase as a class 13 glycoside hydrolase, most similar to pectin acetylesterase, an enzyme involved in cell wall modifications in the peel of a number of fruit. Our results suggest that cell wall hydrolases in tropical fruit may have additional useful roles in biotransforming VOSCs.     

Chemical constituents and antioxidant activity of sweet cherry at different ripening stages

The development and ripening process of sweet cherry (Prunus avium L. cv. 4-70) on the tree was evaluated. For this purpose, 14 different stages were selected in accordance with homogeneous size and color. Some parameters related to fruit quality, such as color, texture, sugars, organic acids, total antioxidant activity, total phenolic compounds, anthocyanins, and ascorbic acid were analyzed. The results revealed that in sweet cherry, the changes in skin color, glucose and fructose accumulation, and softening process are initiated at early developmental stages, coinciding with the fast increase in fruit size. Also, the decrease in color parameter a was correlated with the greatest accumulation of total anthocyanins. Ascorbic acid, total antioxidant activity (TAA), and total phenolic compounds decreased during the early stages of sweet cherry development but exponentially increased from stage 8, which coincided with the anthocyanin accumulation and fruit darkening. TAA showed positive correlations (r(2) = 0.99) with both ascorbic acid and total phenolic compounds and also with the anthocyanin concentration from stage 8. Taking into account the reduced shelf life of sweet cherry and to ensure that these fruits reach consumers with the maximum organoleptic, nutritional, and functional properties, it is advisable to harvest sweet cherries at stage 12 of ripening.     

Antioxidant Metabolism during Fruit Development of Different Acerola ( Malpighia emarginata D.C) Clones

The present research work describes the major changes in the antioxidant properties during development of acerola from five different clones. Ripening improved fruit physicochemical quality parameters; however, total vitamin C and total soluble phenols (TSP) contents declined during development, which resulted in a lower total antioxidant activity (TAA). Despite the decline in TSP, at ripening, the anthocyanin and yellow flavonoid content increased and was mainly constituted of cyanidin 3-rhamnoside and quercetin 3-rhamnoside, respectively. The activities of oxygen-scavenging enzymes also decreased with ripening; furthermore, the reduction in vitamin C was inversely correlated to membrane lipid peroxidation, indicating that acerola ripening is characterized by a progressive oxidative stress. Among the studied clones, II47/1, BRS 237, and BRS 236 presented outstanding results for vitamin C, phenols, and antioxidant enzyme activity. If antioxidants were to be used in the food supplement industry, immature green would be the most suitable harvest stage; for the consumer's market, fruit should be eaten ripe.     

Carotenoid biosynthesis changes in five red pepper (Capsicum annuum L.) cultivars during ripening. Cultivar selection for breeding

Changes in the biosynthesis of individual carotenoid pigments have been investigated during fruit ripening of five cultivars of red pepper (Capsicum annuum L.): Mana, Numex, Belrubi, Delfin, and Negral (a chlorophyll-retaining mutant when ripe). The study was carried out throughout the ripening process, and with special emphasis on the ripe stage, to discover possible differences between cultivars and to characterize these by their carotenoid pattern and content for selecting the best varieties for breeding programs. Ripening fruit of the five cultivars showed the typical and characteristic pattern of carotenoid biosynthesis for the Capsicum genus. In the five cultivars, lutein and neoxanthin, both characteristic chloroplast pigments, decreased in concentration with ripening and eventually disappeared. beta-Carotene, antheraxanthin, and violaxanthin increased in concentration, and other pigments were biosynthesized de novo: zeaxanthin, beta-cryptoxanthin, capsanthin, capsorubin, capsanthin-5,6-epoxide, and cucurbitaxanthin A. A pool of zeaxanthin stands out of the rest of pigment during ripening, which reveals the importance of this pigment as a branching point in the carotenoid biosynthesis in Capsicum. Quantitatively, Negral cultivar showed the highest increase in total carotenoid content (48. 39-fold), followed by Mana and Delfin with 38.03- and 36.8-fold, respectively, and by Belrubi and Numex with 28.03- and 23.48-fold, respectively. In all the red varieties, there was an inverse relationship between total carotenoid content and the red to yellow isochromic pigment fraction ratio (R/Y) and the capsanthin-to-zeaxanthin ratio (Caps/Zeax). This seems to be related to the carotenogenic capacity of the cultivar, and thus selection and breeding should not only seek a higher total carotenoid content but also attempt to increase these ratios. In the present study, the cultivar Mana had the highest total carotenoid content (13 208 mg/kg dwt), but the lowest R/Y (1.25) and Caps/Zeax (3.38) ratios, which are therefore the parameters to improve. The cultivar Negral had a high carotenoid content (8797 mg/kg dwt) and high R/Y and Caps/Zeax ratios and could be used for transfer of these characters in direct crosses with the cultivar Mana. The cultivar Numex had the highest Caps/Zeax ratio (7.17) and is thus an ideal progenitor for this character.