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John R. Stommel and Kathleen G. Haynes

Inheritance of resistance to tomato anthracnose caused by Colletotrichum coccodes (Wallr.) S.J. Hughes was evaluated in parental, F1, F2, and backcross populations developed from crosses between adapted resistant (88B147) and susceptible (90L24) tomato (Lycopersicon esculentum Mill.) breeding lines. Resistance was evaluated via measurement of lesion diameters in fruit collected from field-grown plants and puncture inoculated in a shaded greenhouse. Backcross and F2 populations exhibited continuous distributions suggesting multigenic control of anthracnose resistance. Anthracnose resistance was partially dominant to susceptibility. Using generation means analysis, gene action in these populations was best explained by an additive-dominance model with additive × additive epistatic effects. A broad-sense heritability (H) of 0.42 and narrow-sense heritability (h2) of 0.004 was estimated for resistance to C. coccodes. One gene or linkage group was estimated to control segregation for anthracnose resistance in the cross of 90L24 × 88B147.

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John R. Stommel and Bruce D. Whitaker

Eggplant (Solanum melongena L.) is ranked among the top ten vegetables in terms of oxygen radical absorbance capacity due to its fruit's phenolic constituents. Several potential health promoting effects have been ascribed to plant phenolic phytochemicals. We report here a first evaluation of phenolic acid constituents in eggplant fruit from accessions in the USDA eggplant core subset. The core subset includes 101 accessions of the cultivated eggplant, S. melongena, and 14 accessions representing four related eggplant species, S. aethiopicum L., S. anguivi Lam., S. incanum L., and S. macrocarpon L. Significant differences in phenolic acid content and composition were evident among the five eggplant species and among genotypes within species. Fourteen compounds separated by HPLC, that were present in many but not all accessions, were identified or tentatively identified as hydroxycinnamic acid (HCA) derivatives based on HPLC elution times, UV absorbance spectra, ES-—MS mass spectra, and in some cases proton NMR data. These phenolics were grouped into five classes: chlorogenic acid isomers, isochlorogenic acid isomers, hydroxycinnamic acid amide conjugates, unidentified caffeic acid conjugates, and acetylated chlorogenic acid isomers. Among S. melongena accessions, there was a nearly 20-fold range in total HCA content. Total HCA content in S. aethiopicum and S. macrocarpon was low relative to S. melongena. A S. anguivi accession had the highest HCA content among core subset accessions. Chlorogenic acid isomers ranged from 63.4% to 96% of total HCAs in most core accessions. Two atypical accessions, S. anguivi PI 319855 and S. incanum PI500922, exhibited strikingly different HCA conjugate profiles, which differed from those of all other core subset accessions by the presence of several unique phenolic compounds. Our findings on eggplant fruit phenolic content provide opportunities to improve eggplant fruit quality and nutritive value.

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John R. Stommel and Judith M. Dumm

Violet to black pigmentation of eggplant (Solanum melongena L.) fruit is caused by anthocyanin accumulation. Model systems demonstrate the role of regulatory genes in the control of anthocyanin biosynthesis. Anthocyanin structural gene transcription requires the expression of at least one member of each of three transcription factor families: MYB, MYC, and WD. To determine the molecular genetic basis for anthocyanin pigmentation in eggplant fruit, we used real-time polymerase chain reaction (PCR) to evaluate the expression of anthocyanin biosynthetic (Chs, Dfr, Ans) and regulatory (Myc, Myb B , Myb C , Wd) genes in S. melongena genotypes that produce fruit with dark violet (‘Classic’) or white (‘Ghostbuster’) coloration, respectively. Transcript levels and anthocyanin content were evaluated in fruit at various stages of development ranging from small post-anthesis fruit to full-sized marketable fruit. Anthocyanin content increased 9-fold in developing violet-colored ‘Classic’ fruit, whereas low but detectable concentrations were found in white ‘Ghostbuster’ fruit. Chs, Dfr, and Ans as well as Myb C and Myc transcript levels were significantly higher in ‘Classic’ in comparison with ‘Ghostbuster’ fruit at comparable stages of fruit development with greatest differences observed for Ans transcript levels. Myb C and Myc transcript levels increased in developing ‘Classic’ fruit coincident with increasing anthocyanin content. Myb B and Wd transcript levels were not coordinated with changes in biosynthetic transcript levels or anthocyanin concentration.

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John R. Stommel and Robert J. Griesbach

Considerable diversity exists in Capsicum L. germplasm for fruit and leaf shape, size, and color as well as plant habit. Using F1, F2, and backcross generations developed from diverse parental stocks, this report describes the inheritance patterns and relationships between unique foliar characters and diverse fruit and plant habit attributes. Our results demonstrate that pepper fruit color, shape, and fruit per cluster were simply inherited with modifying gene action. Broad-sense heritability for fruit color and shape and fruit per cluster was high, whereas narrow-sense heritability for these characters was moderate to low. Although fruit clustering was simply inherited, the number of fruit per cluster exhibited a quantitative mode of inheritance. High fruit counts per cluster were linked with red fruit color and anthocyanin pigmented foliage. Fruit shape was linked with immature fruit color and inherited independently of mature fruit color. Leaf color, length, and plant height were quantitatively inherited. Leaf shape did not vary, but leaf length varied and was positively correlated with leaf width. Broad-sense heritability for leaf characters, including leaf length, leaf width, and leaf color, was high. With the exception of leaf width, which exhibited low narrow-sense heritability, high narrow-sense heritability for leaf characters denoted additive gene action. Plant height displayed high broad-sense heritability. Moderate narrow-sense heritability suggested that additive effects also influence plant height. Analysis of segregating populations demonstrated that red and orange fruit color can be combined with all possible leaf colors from green to black. These results provide new data to clarify and extend available information on the inheritance of Capsicum fruit attributes and provide new information on the genetic control of leaf characters and plant habit.

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John R. Stommel, Ruth S. Kobayashi, and Stephen L. Sinden

Somatic fusion hybrids created between tomato and Solanum ochranthum, a wild nontuber-bearing diploid species that is genetically isolated from tomato, were evaluated in an effort to introgress traits from S. ochranthum into tomato. Pollen stainability and pollen tube growth examination demonstrated that little or no viable pollen was present in tetraploid and hexaploid fusion hybrids. Aneuploidy was noted in a small percentage of these hybrids. Use of tetraploid and hexaploid fusion hybrids as female parents in backcrosses to diploid and tetraploid tomato was studied. Chemical treatments that induce either chromosomal recombination or reduction may be advantageous for overcoming difficulties in introgression of these wide hybrids into tomato.

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Ruth S. Kobayashi, Stephen L. Sinden, and John R. Stommel

Incorporation of genes from wild species has been a major contributor to tomato improvement in recent years. Solanum ochranthum, a woody non-tuber bearing species, is a potential source of resistance against tomato diseases and insect pests but is genetically isolated from tomato. Somatic hybridization methods were developed to facilitate the use of S. ochranthum for tomato germplasm improvement. Leaf mesophyll protoplasts of S. ochranthum and a Lycopersicon esculentum hybrid were chemically fused with polyethylene glycol. The protoplasts were initially cultured in Shepard's CL, a MS based medium, containing 1 mg·1-1 NAA, 0.5 mg·1-1 BAP and 0.5 mg·1-1 2,4-D. Hybrid regenerants and regenerants of the L. esculentum parent were recovered; S. ochranthum did not regenerate. Hybridity was established by morphological characters, peroxidase isozyme and RAPD markers. Use of these somatic hybrids for tomato improvement was evaluated.

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Catherine M. Ronning, Lind L. Sanford, and John R. Stommel

Colorado Potato Beetle (Leptinotarsa decemlineata Say., CPB) is a destructive pest of the cultivated potato, Solanum tuberosum L. Certain glycoalkaloids in potato leaves are effective deterrents to this insect; however, in tubers these compounds can be toxic to humans. Leptines are foliar-specific glycoalkaloids produced by the related species, S. chacoense. These compounds have been shown to confer resistance to CPB. We are studying the inheritance of leptine production in segregating F1 and F2 populations derived from two S. chacoense accessions, 55-1 and 55-3, which are (respectively) high and low leptine producers. The F1 segregates 1:1 for high (>70% of total glycoalkaloids) and low (<20% of TGA) leptine content. Segregation data from the F1 and F2 populations suggest a twogene model for leptine production: a dominant repressor and a recessive inducer. Using two bulked DNA samples composed of highand low-leptine individuals from the F1 population, we are using various types of molecular markers (RAPDs, SSRs, DS-PCR, and AFLPs) to search for markers linked to leptine production. We have identified a RAPD band that appears to be closely associated with low leptine content and supports the two-gene model. The use of such a marker in a breeding program will facilitate the development of CPB resistant potato varieties.

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John R. Stommel, Robert W. Goth, and Kathleen G. Haynes

Bacterial soft rot of bell pepper (Capsicum annuum L.), caused by Erwinia spp., is a destructive postharvest market disease of this crop. Control is presently limited to chemical treatments. Methods of inoculating pepper fruit were evaluated to develop a reliable technique for soft rot resistance screening. Erwinia carotovora subsp. atroseptica (Eca) was isolated from partially decayed field grown pepper fruit at Beltsville, MD. Fruit were inoculated with suspensions of Eca via: (a) abrasion with Carborundum, (b) hypodermic puncture, or (c) non-wounded tissue. Inoculated fruit were held under high humidity at 21-23C for two to three days prior to scoring. Degree of soft rot decay was determined via fruit weight loss from two replicates of the experiment over the course of the growing season. Significant differences were not evident among varieties or experiment dates for weight loss due to tissue decay. Hypodermic puncture inoculation was superior to other methods for inducing fruit rot.

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Gordon J. Lightbourn, Robert J. Griesbach, and John R. Stommel

Color observed in plants is due to several pigments, in particular chlorophylls, carotenoids, flavonoids, and betalains. The many hues can be attributed to a number of biochemical factors, inclusive of pigment concentration, pigment combinations and their ratios, and vacuolar pH. Shades of violet to black pigmentation in pepper (Capsicum annuum L.) are attributed to anthocyanin accumulation. The color of unripe pepper fruit varies from green and yellow to ivory, through varying shades of violet and purple to nearly black. Whereas pepper fruit color is important for culinary product quality, foliar pigmentation is also an important aspect of ornamental variety appeal. Foliage and stem color may vary from green to varying shades of green/purple to nearly black. HPLC analysis of violet and black pepper fruit revealed a single anthocyanidin that was identified as delphinidin. Black fruit contained five-fold higher chlorophyll concentrations in comparison to violet fruit, which contained relatively little chlorophyll. Differences in fruit pH were not statistically significant. Similar to fruit, black pepper leaf tissue contained delphinidin as the predominant anthocyanidin, but in higher concentration relative to that found in fruit. The results demonstrate that high concentrations of delphinidin in combination with chlorophyll account for black pigmentation. Real-time PCR analysis of tissues that varied in pigmentation intensity due to varying anthocyanin concentration revealed functional, but differentially expressed, structural genes in the anthocyanin biosynthetic pathway. Analysis of regulatory gene expression identified a MYB transcription factor that was differentially expressed in response to varying anthocyanin concentration.

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John R. Stommel, Judith A. Abbott, and Robert A. Saftner