Journal of Heredity 2003:94(6)
© 2003 The American Genetic Association 94:449-456
Characterization and Inheritance of the Anthocyanin fruit (Aft) Tomato
From the Department of Horticulture, 4017 ALS, Oregon State University, Corvallis, OR 97331. C. M. Jones is currently at the C. M. Rick Tomato Genetics Resource Center, Department of Vegetable Crops, University of California, One Shields Ave., Davis, CA 95616.
Address correspondence to J. R. Myers at the address above, or e-mail: myersja{at}bcc.orst.edu.
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Tomato (Lycopersicon esculentum) accession LA1996 with the Anthocyanin fruit (Aft) gene has dark green foliage, elevated anthocyanin expression in the hypocotyls of seedlings, and anthocyanin in the skin and outer pericarp tissues of the fruit. Interest in the health benefits and antioxidant capacity of anthocyanins led to this study of the genetic potential for increased levels of this important class of phytonutrients in tomato fruit. In order to conform to tomato gene nomenclature rules, we propose changing the symbol Af for Anthocyanin fruit to Aft. Segregation ratios of anthocyanin expression in F2 and BC1 populations of a cross between the processing tomato UC82B and LA1996 were consistent with a single dominant gene hypothesis. Anthocyanin expression was reduced in backcross populations compared to F2 populations. Anthocyanin concentration, as measured by the pH differential method, of pigment-rich pericarp and skin tissues from LA1996 was estimated to be 20.6 mg/100 g and 66.5 mg/100 g, respectively. Anthocyanidin composition was characterized by high-performance liquid chromatography (HPLC). Fruit of accession LA1996 contained predominantly petunidin, followed by malvidin and delphinidinin. Lycopene, ß-carotene, phytoene, and phytofluene levels were similar to those of normal tomatoes and lower than those found in high pigment tomatoes.
Anthocyanins are the most common class of purple, red, and blue plant pigments. More than 300 different anthocyanin compounds have been identified in plants. They are planar molecules with a C6-C3-C6 carbon structure typical of flavonoids. The compounds are separated into distinct classes based on (1) substitution by hydroxyls at positions 3', 4', or 5' of the phenyl "B" ring, (2) the type of glycosidic subunits, and (3) acyl substitution of the sugar hydroxyl (Mazza and Miniati 1993). Anthocyanins have received particular attention because of their very strong antioxidant activity as measured by the oxygen radical absorbing capacity (ORAC) assay. Grapes (Wang et al. 1996), blueberries, blackberries, raspberries, and cherries (Wang et al. 1997) all have high antioxidant capacity in comparison to other fruits and vegetables. Dietary phytonutrients and antioxidants provided by fruits and vegetables are considered to be of primary importance in the prevention of disease and aging (Ames 1983; Ames et al. 1993). Anthocyanins and related polyphenolic and flavonoid compounds are increasingly considered important phytonutrient contributors (Hollman et al. 1996; Lazarus and Schmitz 2000; Lean et al. 1999; Proteggente et al. 2002; Soleas et al. 1997).
Anthocyanin in Tomato Fruit
Tomato (Lycopersicon esculentum) fruit are not usually reported to contain anthocyanin. The red color in tomatoes is from the carotenoid, lycopene (Rick and Stevens 1986). The fruit of several closely related species—Lycopersicon chilense, Lycopersicon hirsutum, Lycopersicon cheesmanii, and Solanum lycopersicoides—do, however, contain anthocyanins (Giorgiev 1972; Rick 1964; Rick et al. 1994). Anthocyanin fruit (Aft) from L. chilense, Aubergine (Abg) from S. lycopersicoides, and atroviolacium (atv) from L. cheesmanii cause anthocyanin expression in tomato fruit.
A single paragraph note reported the presence of the dominant gene, Af (Aft) for Anthocyanin fruit, derived from a L. esculentum x L. chilense cross (Giorgiev 1972). Giorgiev (1972) reported that the purple color was caused by anthocyanins, though scant evidence was presented to confirm the pigment was indeed anthocyanin or verify inheritance of the trait. It is important to note that while Af has been used to represent Anthocyanin fruit, the af gene symbol was already in use for an independent gene, the recessive anthocyanin free, which lacks anthocyanin in vegetative tissues (Burdick 1958; Clayberg 1960). In order to conform to tomato gene nomenclature rules (Clayberg 1970), we propose the gene symbol Aft for Anthocyanin fruit.
The allelic relationships between Aft, Abg, and atv remain unclear. Abg resulted from the intergeneric cross S. lycopersicoides x L. esculentum and is linked to a random amplified polymorphic DNA (RAPD) marker on chromosome 10 (Rick et al. 1994). It is possible that Abg lies in the chromosomal inversion recently identified between S. lycopersicoides and L. esculentum on chromosome 10 (Pertuze et al. 2002). Homozygous S. lycopersicoides introgressions of this region were not achieved using a marker-assisted selection approach (Canady 2002). Stable homozygotes of Abg have not been obtained. The instability of Abg has impeded traditional allele tests with Aft. L. chilense, the donor genome for Aft, is not known to contain any chromosomal inversions in relation to L. esculentum. It is possible that Aft is an allele of Abg and resides on chromosome 10, however, six restriction fragment length polymorphism (RFLP) loci surveyed along chromosome 10 of LA1996 failed to uncover a L. chilense introgression (Jones C, unpublished data). Aft and Abg show differential expression of anthocyanin in and beneath the epidermal cell layer. Plants with the gene atv (derived from a Galapagos island accession described in the original publication as L. pimpinellifolium but now considered to be L. cheesmanii) also express anthocyanin (Rick 1964). Pigment expression in atv is distinct from Aft. Expression in vegetative tissues is more intense in atv, while expression in the fruit is not as great and appears to be expressed in different cell layers not visually detectable (Mes P, unpublished observations) as in Aft.
Reports of pigments attributed to anthocyanins in tomato fruit are confusing. Many heirloom varieties of tomatoes whose names contain "purple" and "black" do not contain anthocyanins. Rather, the green flesh (gf) gene(s) is probably involved (Butler 1962; Kerr 1956), and the muddy brown fruit colors are attributed to persistent green chlorophyll combined with red from lycopene (Jones 2000). (During normal tomato ripening, chlorophyll breaks down concurrent with the increase in carotenoid formation.) However, no allelism tests have been reported between these heirloom varieties and the green flesh gene. The functionally homologous gene cl, inhibiting chlorophyll breakdown, is responsible for green and brown mature fruit color in peppers (Smith 1950).
Anthocyanin in Tomato Vegetative Tissues
Normal tomato genotypes routinely have anthocyanin in vegetative tissues. Previously identified genes (a, aa, ae, ai, al, af, afr, ah, aw, bls, hp-1, and hp-2) are known to affect anthocyanin levels in vegetative tissues. Bovy et al. (2002) identified petunidin 3-(p-coumaryl rutinoside)-5-glucoside and malvadin 3-(p-coumaryl rutinoside)-5-glucoside as the principal anthocyanins in tomato vegetative tissue. This is consistent with vegetative anthocyanins identified by Gromer (2000). Petunidin 3-(p-coumaryl rutinoside)-5-glucoside was tentatively identified in extracts from wild-type seedlings, while peonidin-3-(p-coumaryl rutinoside)-5-glucoside was tentatively identified in tomato mutants a, ag, ai, al, atv, and hp-1 (Von Wettstein-Knowles 1968).
Anthocyanins in Related Solanaceae
Petunidin 3-(p-coumaryl-rutinoside)-5-glucoside is the principal anthocyanin found in garden huckleberry (Solanum scabrum), purple-fleshed potatoes (Solanum tuberosum), as well as the fruit of purple tomatillos (Physallis ixocarpa) (Mazza and Gao 1994; Price and Wrolstad 1995). In contrast, red-fleshed potatoes are characterized as containing predominantly pelargonidin 3-(p-coumaryl rutinoside)-5-glucoside (Rodriguez-Saona et al. 1998). Eggplant (Solanum melongena) contains principally delphinidins (Jackman and Smith 1996).
Rational
High antioxidant activity associated with anthocyanins in other species suggests that increasing anthocyanin content in tomato fruit may increase their antioxidant capacity. Recent attempts to enhance flavonoid and antioxidant levels through transgenic expression of chalcone isomerase (Muir et al. 2001; Verhoeyen et al. 2002) and maize transcription factors LC and C1 (Bovy et al. 2002) have dramatically increased flavonoid levels, though not anthocyanins, in tomato fruit.
Plants containing the Aft gene have phenotypic similarities to tomatoes with the high pigment genes. The high pigment tomato fruits have high levels of carotenoids, flavonoids, and vitamin C due to a mutation that exaggerates phytochrome response (Barker and Tomes 1964; Kerckhoffs et al. 1997; Kerr 1957, 1965; Minoggio et al. 2003; Palmieri et al. 1978; Peters et al. 1992). The homologous phenotypes include deeper green foliage and stems, as well as intense anthocyanin expression in the hypocotyls and roots of germinating seedlings. Pigment production in Aft fruit is stimulated by light exposure (Giorgiev 1972) and phytochromes are known to act in light-mediated regulation of anthocyanin production (Adamse et al. 1989; Kerckhoffs and Kendrick 1993). Consequently phytochrome action may be involved in the Anthocyanin fruit phenotype, and other physiological traits may be affected as they are in high pigment tomatoes.
The objectives of this study were to verify the inheritance of Aft, describe its expression, and measure total anthocyanin and carotenoid levels.
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Materials and Methods |
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Germplasm Sources
The accession LA1996 containing Aft (Figure 1) was obtained from the C. M. Rick Tomato Genetics Resource Center, Davis, CA. LA1996 was originally contributed and annotated as containing Anthocyanin fruit by Giorgiev (1972). UC82B, a red-fruited open-pollinated processing tomato, was provided by Lockhart Seed Company, Stockton, CA.
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Anthocyanin Concentration
Ripe tomatoes were harvested from five plants grown at the Seeds of Change Research Farm, Corvallis, OR, in 1998. We chose fruit expressing medium to high levels of anthocyanin for extraction in order to represent potential, not average, anthocyanin production. Areas of skin containing anthocyanin were removed with a scalpel and pericarp tissue containing anthocyanins was separated from the rest of the fruit with a vegetable peeler. Pericarp and skin were kept separate and frozen at –23°C until extraction.
We extracted anthocyanins according to the procedures of Giusti and Wrolstad (1996). Tissues were frozen with liquid nitrogen and powdered in a mill prior to extraction. A total of 17.64 g of pericarp tissue powder and 6.42 g of skin powder were extracted separately with 100% acetone and filtered with a Buchner funnel. Filter cake was reextracted with 70% acetone. We partitioned the extractant with chloroform in a separatory funnel and allowed it to stand at 1°C for 12 h. The chloroform fraction was discarded and the aqueous fraction was rotoevaporated at 40°C until dry and brought to total volume of 50 ml with distilled water. Total anthocyanin content was measured by the pH differential method (Fuleki and Francis 1968) as described by Wrolstad (1976) using a Varian DMS double-beam spectrophotometer. This method estimates total anthocyanin content based on the reversible conversion of anthocyanins from the oxonium to the hemiketal form, but does not determine the individual compounds present. Samples were diluted 5:1 in pH 1.0 and pH 4.5 buffer. The difference in absorption between the ultraviolet (UV)-visible adsorption maxima (540 nm for skin tissue and 535.5 for pericarp) and adsorption at 700 nm was determined at pH 1.0 and pH 4.5. The difference between these two values was used to determine the total anthocyanin concentration according to the formula (Abs pH 1.0 – Abs pH 4.5)/
L x 103 x MW x dilution factor. The results are expressed as petunidin-3-(p-coumaroyl rutinoside)-5-glucoside based on 17,000 as the extinction coefficient () and 934, the molecular weight (MW) (Price and Wrolstad 1995). L is the path length (in centimeters) of the spectrophotometer.
High-Performance Liquid Chromatography (HPLC) Anthocyanidin Analysis
We extracted anthocyanins from fruit of accession LA1996 using the method described by Rodriguez-Saona et al. (1998). This method isolates anthocyanins based on the preferential solubility of anthocyanins in acetone versus chloroform. Twelve grams of skin and pericarp tissue containing anthocyanin were excised from the fruit and homogenized in liquid nitrogen for subsequent anthocyanin isolation. Tubes of isolated anthocyanins were stored at –23°C. Anthocyanidins were produced from the isolated anthocyanin solution using the acid hydrolysis protocol for preparation of anthocyanidins (Durst and Wrolstad 2001). Forty milliliters of isolated anthocyanins were thawed and filtered through a 0.45 µm filter attached to an activated C18 SPE cartridge. The SPE cartridge was rinsed twice with 5 ml of acidified water. The anthocyanin was then eluted into a rotoevaporation boiling flask using two washes with 5 ml methanol. The solution was evaporated until a small amount of liquid remained. The solution was placed in a screw-top test tube, to which 10 ml 2 N HCl was added. The tube was flushed with nitrogen, sealed loosely, and placed in a boiling water bath for 30 min. The tube was then placed in an ice bath until cool. The sample was reapplied to a C18 SPE cartridge, rinsed, and rotoevaporated to remove methanol. Two milliliters of 4% phosphoric acid was added and the aliquot was analyzed promptly by HPLC. A sample of the isolated anthocyanins was also saponified using the method described by Durst and Wrolstad (2001) to remove acylation. A subsample of the saponified anthocyanins was subjected to acid hydrolysis and then analyzed by HPLC.
HPLC Carotenoid Analysis
High-performance liquid chromatography analysis was performed using a rapid HPLC method developed for tomato carotenoid analysis (Jones 2000). Homogenized tomato samples were extracted with tetrahydrofuran, filtered, and combined 1:1 with eluent A (85% acetonitrile, 10% methanol, 2.5% hexane, and 2.5% dichloromethane, v:v). Samples were microfiltered with a 0.2 µm nylon syringe filter (Alltech Inc., Deerfield, IL) and 50 µl was injected into the HPLC system. All extractions were carried out under subdued ambient light. HPLC was performed with a Beckman model 334 gradient liquid chromatograph (Beckman Instruments Inc., Berkeley, CA) equipped with a Waters 991 photodiode array detector (Waters Inc., Milford, MA). Analysis was carried out using a reverse phase Spheri-5 RP-18 column (220 mm x 4.6 mm, 5 µm; Perkin Elmer Brownlee, Norwalk, CT) coupled to an ODS-5S guard column (3.0 cm x 4.6 mm; Bio-Rad, Richmond, CA). Gradient conditions employed for the mobile phase consisted of eluent A (85% acetonitrile, 10% methanol, 2.5% hexane, and 2.5% dichloromethane, v:v) and eluent B (45% acetonitrile, 10% methanol, 22.5% hexane, and 22.5% dichloromethane v:v). The chromatographic conditions at a flow rate of 1 ml/min were: 0–1 min, 100% isocratic eluent A; 1–7 min, linear gradient from 0–100% eluent B; and 7–20 min, 100% eluent B.
Genetic Studies
Single plants of LA1996 and UC82B were crossed in the greenhouse during the winter of 1999 by standard emasculation and crossing techniques. F1 plants were grown and observed at the Seeds of Change Research Farm in Corvallis, OR, in 1999. Plants were allowed to self-pollinate and were crossed to UC82B to produce the F2 and the BC1F1 populations. F2 and backcross populations were grown and scored in the greenhouses of Oregon State University (OSU) in the winter/spring of 2000 or at the OSU Vegetable Research Farm in Corvallis, OR, in the summer of 2000 (F2 population only). Greenhouse plants were grown in 4 L pots with a soil-free potting mix (Rexius Forest Products, Eugene, OR) and fertilized with Osmocote 14-14-14 slow release fertilizer (Scott Sierra, Marysville, OH). Natural light was supplemented with high-intensity metal halide and sodium lamps suspended 1.3 m above the bench. Supplementary lighting was provided 10 h/day and temperatures were maintained at a minimum of 24°C during the day and 18°C at night. We evaluated segregating populations in a green fruit stage on a scale of 1–4 (1 represents no anthocyanin and 2, 3, and 4 represent low, medium, and high levels of expression, respectively; see Figure 1). Anthocyanin expression is correlated in the green and red stages; however, we scored in the green fruit stage because low expression levels in some plants are more difficult to score in the presence of mature fruit color. All fruit on the plant was inspected and the plant was scored according to the maximum anthocyanin expression in the fruit.
We analyzed data by using chi-square tests of goodness-of-fit, with the Yates continuity correction recommended for data with one degree of freedom (Little and Hills 1978; Yates 1934). Data from separate populations were pooled for analysis based on a chi-square test of homogeneity (Little and Hills 1978).
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Phenotypic Description
Vegetative tissues of Aft tomato plants are distinctive. Leaves are darker green and stems contain visibly more purple speckling than do wild-type plants. The plants are vigorous and do not have the reduced growth associated with high pigment genotypes (Sayama 1979; Sayama and Tigchelaar 1985; Wann 1996; Wann et al. 1985). With the exception of anthocyanin expression, LA1996 is phenotypically a normal L. esculentum plant, suggesting most of the L. chilense genome has been eliminated. Fruit anthocyanin expression was strongest in areas exposed to light. For example, anthocyanin was absent in areas shaded by the calyx. Anthocyanin expression was strongest in the skin and pericarp tissues beneath the skin (Figure 2). The fruit interior did not contain visible anthocyanin.
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Anthocyanidin Pigments and Anthocyanin Concentration
The purple color of LA1996 (Figures 1 and 2) is produced by the glycosylated anthocyanidins of petunidin, malvidin, and delphinidin, as determined by comparison of elution times of anthocyanidins compared to known standards (Figure 3). Of these, petunidin and malvidin are predominant. The three major peaks of the saponified anthocyanins were not positively identified, but were consistent with the typical elution time of a 3,5-diglycoside, such as the petunidin 3-(p-coumaryl)rutinoside-5-glucoside reported in tomato vegetative tissues (Von Wettstein-Knowles 1968). This would suggest that the anthocyanins present in Aft fruit are petunidin, malvidin, and delphinidin 3-(p-coumaryl-rutinoside)-5-glucoside, though analysis by liquid chromatography-mass spectrophotometry will be necessary to confirm this. Petunidin 3-(p-coumaryl-rutinoside)-5-glucoside is the dominant anthocyanin in garden huckleberry (S. scabrum) fruit (Francis and Harborne 1966; Price and Wrolstad 1995), potatoes (Mazza and Gao 1994), and tomatillo fruit (Price and Wrolstad 1995), which indicates that this compound is widely distributed in the Solanaceae.
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The total monomeric anthocyanin concentration of pigment-rich tissues separated from whole LA1996 fruits was estimated by the pH differential method to be 20.6 mg/100 g in the pericarp tissue and 66.5 mg/100 g in the skin expressed as petunidin 3-(p-coumaryl-rutinoside)-5-glucoside. This demonstrates the anthocyanin production potential from pigment-rich tissues, not the average values for LA1996 fruit, as only pigment-rich tissues were extracted. Concentrations of anthocyanins measured by this method may be affected by copigmentation with other flavonoids (Asen 1975; Asen et al. 1972). Very little copigmentation occurs at pH 2.0 or less, but increases with pH (Asen et al. 1972). We measure absorbance at pH 1.0 and subtract the pH 4.5 value; therefore if copigmentation occurs, greater absorbance at pH 4.5 would cause us to underestimate actual concentrations.
Inheritance of Aft
The hypothesis that a single dominant gene confers the Anthocyanin fruit phenotype is consistent with our data. F1 progeny of 12 separate pollinations to LA1996 were evaluated in the field in 1999, and all F1 plants expressed anthocyanin in the fruit. We subjected the three F2 populations (derived from three separate F1 plants representing different pollination events) to a homogeneity test (P =.601) and pooled the data (Table 1). F2 progeny of a cross between LA1996 (Aft) and the processing cultivar UC82B were consistent with the hypothesis of a single dominant gene for Anthocyanin fruit (P =.190). We tested two gene ratios of 9:7 and 13:3 and found the data fit neither model. Progeny of two different backcrosses to UC82B segregated in a manner consistent with a single dominant gene hypothesis when analyzed separately (Table 1). However, after a test of homogeneity and pooling of data from progeny of both backcross populations, there were fewer than expected Anthocyanin fruit (Aft aft) plants (P =.034). This may have resulted from Aft deriving from an interspecific cross between L. esculentum and L. chilense for which significant fertility barriers exist (Rick and Stevens 1986). Elimination of an allele through chromosome loss could be involved. Alternatively, lower expression of anthocyanin in the backcross population than in the F2 could result from reduced expression in heterozygous BC1F1 plants or result from lower light levels in the greenhouse environment where the backcross populations were grown. Consequently some heterozygous plants could have displayed such limited expression that they were incorrectly scored.
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Chi-square tests are presented (Table 1) based on the presence or absence of anthocyanin. Levels of anthocyanin expression were variable and were scored from 1 (no anthocyanin) to 4 (high anthocyanin). The Aft phenotype differed between the F2 population and the backcross populations, with very little medium or high anthocyanin expression in the backcrosses (Figure 4). While the low anthocyanin levels in the backcross populations as compared to the F2 generation may suggest reduced expression in the heterozygotes, we did not grow F3 plants from putative heterozygotes to address the possibility of incomplete dominance.
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Normal tomato genotypes routinely contain anthocyanin in the vegetative parts of the plants but not in the fruit. The cuticles of L. esculentum fruit contain chalconaringenin, the pigment believed to be responsible for the yellow color of tomato skin (Hunt and Baker 1980). Chalcones are biosynthetic precursors to the anthocyanins. These facts indicate that all the structural genes necessary for anthocyanin production are already present in the tomato genome. Consequently we believe Aft is more likely a regulatory gene or promoter region of a structural gene that up-regulates fruit anthocyanin expression. Bovy et al. (2002) found that transgenic fruit with high levels of flavonoids did not produce anthocyanins due to low fruit expression of flavonoid 3'-hydroxylase and flavonoid 3'5'-hydroxylase. It is likely that these genes are up-regulated in Aft fruit.
While Aft has phenotypic similarities to the hp phytochrome response mutants, Aft may be unrelated to the phytochrome response mechanism. While the vegetative and fruit characteristics share similarities, the deep vegetative and fruit color of the hp mutants is considered to be principally the result of increased chlorophyll in the leaves and unripe fruit and carotenoid content in ripe fruit (Barker and Tomes 1964; Jarret et al. 1984; Wann et al. 1985). In contrast, we attribute the dark foliage and dark ripe fruit characteristic of LA1996 to the presence of anthocyanins. In addition, unripe fruit of LA1996 are not dark green like that of the phytochrome mutants, and we demonstrate in this article no carotenoid increase in the ripe fruit. Also lacking in the Anthocyanin fruit tomato plants are the brittle stems and characteristic shortened seedling hypocotyls found in the hp mutants (Jarret et al. 1984).
Carotenoids of the Anthocyanin fruit Tomato
The phenotypic similarities between the Aft tomato plants and high pigment tomato plants led us to investigate the possibility that LA1996 may also have heightened carotenoid levels (Jones 2000). Carotenoid levels observed in LA1996 in three separate trials were not statistically different from normal tomatoes. The mean carotenoid content of LA1996 was 1.93 mg/100 g FW phytoene, 1.31 mg/100 g FW phytofluene, 0.64 mg/100 g FW ß-carotene, and 5.15 mg/100 g FW lycopene. Aft fruit were significantly lower in lycopene and ß-carotene than hp-1, hp-2, and dg genotypes. While nearly isogenic lines with Aft are not yet available, our results indicate that it is unlikely that Aft substantially alters carotenoid quantity or carotenoid ratios.
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We have established the presence and quantity of anthocyanins in Aft tomatoes. We have identified petunidin, followed by malvidin and delphinidinin as the principal anthocyanidins in Anthocyanin fruit. We have shown that the Anthocyanin fruit phenotype introgressed into L. esculentum from L. chilense is inherited in a manner consistent with a single dominant gene. Aft plants have shared phenotypic traits with other tomato photomorphogenic mutants (hp-1, hp-2, and dg). In contrast to these high pigment tomatoes, no increase in carotenoid content was associated with the presence of Aft. While intense anthocyanin expression is needed for strong antioxidant activity, the introduction of the Anthocyanin fruit characteristic into carotenoid-rich tomatoes provides the opportunity to develop new cultivars rich in water- and lipid-soluble antioxidants. The simple inheritance of Aft makes utilization of this gene in existing tomato germplasm feasible.
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Acknowledgments |
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We gratefully acknowledge the laboratory of Ron Wrolstad, especially Robert Durst, for assistance with the anthocyanin analysis. We wish to thank the C. M. Rick Tomato Genetics Resource Center and Lockhart Seed Co. for seeds used in this study. This work was supported by Seeds of Change Inc. and the Vegetable Breeding Program at Oregon State University.
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Footnotes |
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Corresponding Editor: Reid G. Palmer
Received November 8, 2002
Accepted July 31, 2003
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Adamse P, Peters JL, Jaspers PA, van Tuinen A, Koornneef M, Kendrick RE, 1989. Photocontrol of anthocyanin synthesis in tomato seedlings: a genetic approach. Photochem Photobiol. 50:107-111.
Ames BN, 1983. Dietary carcinogens and anticarcinogens. Science. 221:1256-1264.
Ames BN, Shigenaga MK, Hagen TM, 1993. Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci USA. 90:7915-7922.
Asen S, 1975. Factors affecting flower color. Acta Hort. 41:57-68.
Asen S, Stewart RN, Norris KH, 1972. Co-pigmentation of anthocyanins in plant tissues and its effect on color. Phytochemistry. 11:1139-1144.[CrossRef]
Barker LR, Tomes ML, 1964. Carotenoids and chlorophylls in two tomato mutants and their hybrid. Proc Am Soc Hort Sci. 85:507-513.
Bovy A, de Vos R, Kemper M, Schijlen E, Almenar Pertejo M, Muir S, Collins G, Robinson S, Verhoeyen M, Hughes S, Santos-Buelga C, van Tuinen A, 2002. High-flavonol tomatoes resulting from the heterologous expression of the maize transcription factor genes LC and C1. Plant Cell. 14:2509-2526.
Burdick AB, 1958. New mutants. Rep. Tomato Genet. Coop.. 8:9-11.
Butler L, 1962. Muddy fruit, a phenotype of the gene gf. Rep. Tomato Genet. Coop.. 12:19.
Canady M, 2002. Development and genetic studies of a Solanum lycopersicoides introgression line library (PhD dissertation). Davis: University of California.
Clayberg CD, 1960. The report of the gene list committee. Rep. Tomato Genet. Coop.. 10:3-.
Clayberg CD, 1970. Rules for nomenclature in tomato genetics. Rep. Tomato Genet. Coop.. 20:3-.
Durst RW, Wrolstad R, 2001. Separation and characterization of anthocyanins by HPLC. In: Current protocols in food analytical chemistry. (Wrolstad R ed). New York: Wiley. F1.3.1–F1.3.13.
Francis FJ, Harborne JB, 1966. Anthocyanins in the garden huckleberry, Solanum guineese. J Food Sci. 31:524-528.[CrossRef]
Fuleki T, Francis FJ, 1968. Quantitative determination of anthocyanins 2. Determination of total anthocyanin and degradation index for cranberry juice. J Food Sci. 33:78-83.[CrossRef]
Giorgiev C, 1972. Anthocyanin fruit tomato. Rep. Tomato Genet. Coop.. 22:10.
Giusti MN, Wrolstad RE, 1996. Characterization of red radish anthocyanins. J Food Sci. 61:322-326.[CrossRef]
Gromer KC, 2000. Determination of anthocyanin structure in VFN8 and Dgt tomato varieties by HPLC (B.S. thesis). San Luis Obispo: California Polytechnic State University.
Hollman PCH, Hertog MGL, Katan MB, 1996. Role of dietary flavonoids in protection against cancer and coronary heart disease. Biochem Soc Trans. 24:785-789.[Web of Science][Medline]
Hunt GM, Baker EA, 1980. Phenolic constituents of tomato fruit cuticles. Phytochemistry. 19:1415-1419.
Jackman RL, Smith JL, 1996. Antocyanins and betalains. In: Natural food colorants, 2nd ed (Hendry GAF and Houghton JD, eds). London: Blackie.
Jarret RL, Sayama H, Tigchelaar EC, 1984. Pleiotropic effects associated with the chlorophyll intensifier mutations high pigment and dark green in tomato. J Am Soc Hort Sci. 109:873-878.
Jones CM, 2000. Evaluation of carotenoids and anthocyanins in high pigment, processing, heirloom and anthocyanin fruit tomatoes (Masters thesis). Corvallis: Oregon State University.
Kerckhoffs LHJ, Kendrick RE, 1993. Photocontrol of anthocyanin biosynthesis in tomato. J Plant Res Tokyo. 110:141-149.
Kerckhoffs LHJ, Schreuder MEL, van Tuinen A, Koornneef M, Kendrick RE, 1997. Phytochrome control of anthocyanin biosynthesis in tomato seedlings: analysis using photomorphogenic mutants. Photochem Photobiol. 65:374-381.
Kerr EA, 1956. Green flesh, gf. Rep. Tomato Genet. Coop.. 6:17.
Kerr EA, 1957. The effect of high pigment genes hp-1, and hp-2, on certain plant constituents. Rep. Tomato Genet. Coop.. 7:9.
Kerr EA, 1965. Identification of high-pigment tomatoes in the seedling stage. Can J Plant Sci. 45:104-105.
Lazarus S, Schmitz H, 2000. Dietary flavonoids may promote health, prevent heart disease. Calif Agric. 54:33-39.
Lean ME, Noroozi M, Kelly I, Burns J, Talwar D, Sattar N, Crozier A, 1999. Dietary flavonols protect diabetic human lymphocytes against oxidative damage to DNA. Diabetes. 48:176-81.[Abstract]
Little TM, Hills FJ, 1978. Agricultural experimentation: design and analysis. New York: Wiley.
Mazza G, Gao L, 1994. Acylated anthocyanins in purple hulls of sunflower seeds and purple fleshed potatoes. In: Proceedings of the first international symposium on natural colorants (Hereld PC ed). Hamden, Connecticut: SIC Publishing.
Mazza G, Miniati E, 1993. Anthocyanins in fruits, vegetables, and grains. Boca Raton, FL: CRC Press.
Minoggio M, Bramati L, Simonetti P, Gardana C, Iemoli L, Santangelo E, Mauri PL, Spigno P, Soressi GP, Pietta PG, 2003. Polyphenol pattern and antioxidant activity of different tomato lines and cultivars. Ann Nutr Metab. 47:64-69.[CrossRef][Web of Science][Medline]
Muir SR, Collins GJ, Robinson S, Hughes S, Bovy A, De Vos CHR, van Tuinen AJ, Verhoeyen ME, 2001. Overexpression of petunia chalcone isomerase in tomato results in fruit containing increased levels of flavonols. Nat Biotechnol. 19:470-474.[CrossRef][Web of Science][Medline]
Palmieri S, Martiniello P, Soressi GP, 1978. Chlorophyll and carotene content in high pigment and green flesh fruits. Rep. Tomato Genet. Coop.. 28:10.
Pertuze RA, Ji Y, Chetelat RT, 2002. Comparative linkage map of the Solanum lycopersicoides and S. sitiens genomes and their differentiation from tomato. Genome. 45:1003-1012.[Medline]
Peters JL, Schreuder MEL, Verduin SJW, Kendrick RE, 1992. Physiological characterization of a high-pigment mutant of tomato. Photochem Photobiol. 56:75-82.
Price CL, Wrolstad RE, 1995. Anthocyanin pigments of Royal Okanogan huckleberry juice. J Food Sci. 60:369-374.[CrossRef]
Proteggente AR, Pannala AS, Paganga G, Van Buren L, Wagner E, Wiseman S, Van De Put F, Dacombe C, Rice-Evans CA, 2002. The antioxidant activity of regularly consumed fruit and vegetables reflects their phenolic and vitamin C composition. Free Radic Res. 36:217-233.[CrossRef][Web of Science][Medline]
Rick C, Stevens AM, 1986. Genetics and breeding. In: The tomato crop: a scientific basis for improvement (Atherton JG and Rudick J, eds). New York: Chapman and Hall; 35–105.
Rick CM, 1964. Biosystematic studies on Galapagos Island tomatoes. Occas Paper Calif Acad Sci. 44:59.
Rick CM, Cisneros P, Chetelat RT, Deverona JW, 1994. Abg, a gene on chromosome 10 for purple fruit derived from S. lycopersiciodes. Rep. Tomato Genet. Coop.. 44:29-30.
Rodriguez-Saona LE, Giusti MM, Wrolstad RE, 1998. Anthocyanin pigment composition of red-fleshed potatoes. J Food Sci. 63:458-465.[CrossRef]
Sayama H, 1979. Morphological and physiological effects associated with the crimson (ogc), high pigment (hp) and other chlorophyll intensifier genes in tomato (Lycopersicon esculentum Mil.) (PhD dissertation). West Lafayette, IN: Purdue University.
Sayama H, Tigchelaar EC, 1985. Yield and processing quality attributes associated with the high pigment (hp) and crimson (ogc) genes in tomato (Lycopersicon esculentum Mill.). Jpn J Breed. 35:145-152.
Smith PG, 1950. Inheritance of brown and green mature fruit color in peppers. J Hered. 41:138-140.
Soleas GJ, Diamandis EP, Goldberg DM, 1997. Resveratrol: a molecule whose time has come? And gone? Clin Biochem. 30:91-113.[CrossRef][Web of Science][Medline]
Verhoeyen ME, Bovy A, Collins G, Muir S, Robinson S, De Vos CH, Colliver S, 2002. Increasing antioxidant levels in tomatoes through modification of the flavonoid biosynthetic pathway. J Exp Bot. 53:2099-2106.
Von Wettstein-Knowles P, 1968. Mutations affecting anthocyanin synthesis in the tomato. Hereditas. 60:317-346.
Wang H, Cao G, Prior RL, 1996. Total antioxidant capacity of fruits. J Agric Food Chem. 44:701-705.[CrossRef]
Wang H, Cao G, Prior R, 1997. Oxygen radical absorbing capacity of anthocyanins. J Agric Food Chem. 45:304-309.[CrossRef]
Wann EV, 1996. Physical characteristics of mature green and ripe tomato fruit tissue of normal and firm genotypes. J Am Soc Hort Sci. 121:380-383.
Wann EV, Jourdain EL, Russel RB, 1985. Effect of mutant genotypes hp ogc and dg ogc on tomato fruit quality. J Am Soc Hort Sci. 110:212-215.
Wrolstad RE, 1976. Color and pigment analyses in fruit products. Station bulletin 624. Corvallis, OR: Agricultural Experiment Station Oregon State University.
Yates F, 1934. Contingency tables involving small numbers and the 
2 test. J R Stat Soc Suppl. 1:217.[CrossRef]
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