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Dean A. Kopsell, Scott McElroy, Carl Sams, and David Kopsell

Vegetable crops can be significant sources of nutritionally important dietary carotenoids and Brassica vegetables are sources that also exhibit antioxidant and anticarcinogenic activity. The family Brassicaceae contains a diverse group of plant species commercially important in many parts of the world. The six economically important Brassica species are closely related genetically. Three diploid species (B. nigra, B. rapa, and B. oleracea) are the natural progenitors of the allotetraploid species (B. juncea, B. napus, and B. carinata). The objective of this study was to characterize the accumulation of important dietary carotenoid pigments among the genetically related Brassica species. The HPLC quantification revealed significant differences in carotenoid and chlorophyll pigment accumulation among the Brassica species. Brassica nigra accumulated the highest concentrations of lutein, 5,6-epoxy lutein, violaxanthin, and neoxanthin. The highest concentrations of beta-carotene and total chlorophyll were found in B. juncea. Brassica rapa accumulated the highest concentrations of zeaxanthin and antheraxanthin. For each of the pigments analyzed, the diploid Brassica species accumulated higher concentrations, on average, than the amphidiploid species. Brassicas convey unique health attributes when consumed in the diet. Identification of genetic relationships among the Brassica species would be beneficial information for improvement programs designed to increase carotenoid values.

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David R. Corbin, Frederick J. Perlak, David A. Fischhoff, John T. Greenplate, Zhen Shen, and John P. Purcell

Genetically modified potato and cotton crops that express insecticidal proteins from Bacillus thuringiensis (Bt) have recently been commercialized. These crops display autonomous resistance to specific insect pests, and thus offer major agricultural and environmental benefits. We have implemented a microbial screening program to discover new types of insecticidal proteins for use in transgenic crops. New proteins with diverse modes of action offer opportunities to control insect pests that are not susceptible to Bt insecticidal proteins and to delay or prevent the potential occurrence of resistance of insects to crops genetically modified with Bt genes. Cholesterol oxidase emerged from our screen as a new insecticidal protein with potent activity against the cotton boll weevil. Cholesterol oxidase was acutely toxic to boll weevil larvae, with an LC50 of 2–6 parts per million when ingested in artificial diet feeding assays, and caused marked reductions in fecundity when ingested by adult boll weevils. Cholesterol oxidase also exerted significant, though less severe, toxicity against several lepidopteran pests. The insecticidal action of cholesterol oxidase appears to be due to oxidation of midgut epithelial membrane cholesterol followed by membrane disruption. A cholesterol oxidase gene was cloned and expressed in transgenic tobacco plants to yield plant tissue that exerted potent activity against boll weevil. Expression of this cholesterol oxidase gene in cotton plants may offer significant protection against the cotton boll weevil and may also aid in the mitigation of resistance of cotton lepidopteran pests to Bt proteins.

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Marilyn Rivera-Hernández, Linda Wessel-Beaver, and José X. Chaparro

Squash and pumpkins (Cucurbita sp.) are important contributors of beta-carotene to the diet. Consumers of tropical pumpkin and butternut squash (both C. moschata Duchesne) prefer a deep orange mesocarp color. Color intensity is related to carotene content. Among the five domesticated Cucurbita species, C. moschata and C. argyrosperma Huber have a close relationship. In crosses between these two species, fertile F1 plants can be easily obtained when using C. argyrosperma as the female parent. This research studied the relationship between and within C. moschata and C. argyrosperma by sequencing three genes in the carotenoid biosynthesis pathway and generating gene trees. Genotypes used in the study differed in flesh color from very pale yellow to dark orange. In some cases, haplotypes were associated with a particular mesocarp color. Further study of these types of associations may improve our understanding of color development in Cucurbita. The frequency of single nucleotide polymorphisms (SNPs) in the sequenced fragments was low. There were more SNPs and more heterozygotes among C. moschata accessions than among C. argyrosperma accessions. Haplotypes of the outgroups (C. ficifolia C.D. Bouché and C. maxima Duchesne) were always distinct from C. moschata and C. argyrosperma. These later species had both distinct haplotypes and shared haplotypes. Haplotypes shared among species tended to be maintained in the same branch of the phylogenetic tree, suggesting either gene flow between the species or a common ancestral gene. Both explanations suggest a close genetic and evolutionary relationship between C. moschata and C. argyrosperma.

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Judith A. Abbott*

Demand for fresh fruits and vegetables is increasing worldwide in response to health concerns, wealth, and the desire for variety in the diet. However, consumption of produce is contingent on the ability of the industry to provide high quality fresh produce and on its convenience, as well as on consumer education and economics. Texture measurement is accepted by horticultural industries as a critical indicator of quality of fruits and vegetables. The fresh produce industry and, indirectly, consumers need methods for measuring produce texture to ensure the quality within a grade, and scientists need measurements to quantify the results of their treatments, whether treatments are genetic, chemical, or physical. The variety of attributes required to fully describe textural properties can only be fully measured by sensory evaluation by a panel of trained assessors. However, instrumental measurements are preferred over sensory evaluations for both commercial and research applications because instruments are more convenient, less expensive, and tend to provide consistent values when used by different people. Thus, instrumental measurements need to be developed that predict sensory evaluations of texture. Such instrumental measurements can then provide a common language among researchers, producers, packers, regulatory agencies, and customers. We compare sensory evaluations of specific critical textural attributes to instrumental force/deformation measurements on a wide variety of fruits and vegetables with relatively uniform bulk tissues, such as apples, bananas, carrots, jicama, melons, pears, potatoes, rutabagas, and several others.

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Ji Tian, Zhen-yun Han, Li-ru Zhang, Ting-Ting Song, Jie Zhang, Jin-Yan Li, and Yuncong Yao

Anthocyanins are protective pigments that accumulate in plant organs such as fruits and leaves, and are nutritionally valuable components of the human diet. There is thus considerable interest in the factors that regulate synthesis. Malus crabapple leaves are rich sources of these compounds, and in this study we analyzed leaf coloration, anthocyanin levels, and the expression levels of anthocyanin biosynthetic and regulatory genes in three crabapple cultivars (Royalty, Prairifire, and Flame) following various temperature treatments. We found that low temperatures (LTs) promoted anthocyanin accumulation in ‘Royalty’ and ‘Prairifire’, leading to red leaves, but not in ‘Flame’, which accumulated abundant colorless flavonols and retained green colored leaves. Quantitative reverse transcript PCR (RT-PCR) analyses indicated that the expression of several anthocyanin biosynthetic genes was induced by LTs, as were members of the R2R3-MYB, basic helix–loop–helix (bHLH) and WD40 transcription factor families that are thought to act in a complex. We propose that anthocyanin biosynthesis is differentially regulated in the three cultivars by LTs via the expression of members of this anthocyanin regulatory complex.

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Thomas C. Koch* and Irwin L. Goldman

Carotenoids (provitamin A) and tocopherols (vitamin E) are powerful antioxidants in plants and in the human diet. Carrot (Daucus carota) has been selected for increased levels of carotenoids, contributing to its orange color and reported health benefits. Selection for increased tocopherol has shown success in seed oils, but little progress has been made in the edible portions of most vegetable crops. HPLC measurement following a simultaneous heptane extraction of both compounds has shown a significant (P ≤ 0.001) positive correlation of α-tocopherol with α-carotene (r = 0.65) and β-carotene (r = 0.52). To increase both the tocopherols and carotenoids in plants, 3 populations have been established from select open-pollinated varieties grown in 2002. These populations consist of half-sib families with these differing selection schemes: based strictly on increased α-tocopherol levels; an index to increase α-carotene, β-carotene and α-tocopherol; and a random population in which no selection is occurring. After one cycle of selection, populations were grown on muck soil during the summer of 2003. Compared with the random population, an increase of 24.68% in α-tocopherol concentration was recorded for the population selected strictly on α-tocopherol while increases of 8.47% in α-tocopherol, 9.31% in α-carotene and 7.31% in β-carotene were recorded for the population with index selection. The continuation of these carrot populations shows promise to produce carrot germplasm with improved human nutritive value.

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Jennifer L. Waters and Stephen R. King

Carotenoids are important phytochemical components of our diet and have gained recent attention as important nutritive compounds found mainly in fruits and vegetables with red, orange, and yellow hues. Lycopene is often cited as being inversely correlated with the occurrence of various cancers, in lowering rates of cardiovascular disease, and improving other various other immune responses. Antioxidant activity, specifically oxidative radical quenching power, is the putative rationale for carotenoids' involvement in disease risk reduction. It is unlikely, however, that carotenoid content and antioxidant capacity are directly correlated in the whole food since there are other antioxidants present in watermelon, such as various free amino acids. A total measure of antioxidant potential may prove to be a useful tool for measuring watermelon nutritional value and implementing pursuant breeding goals. One assay that has gained recent popularity is the oxygen radical absorbance capacity (ORAC) assay. ORAC includes two assays that separate lipophylic and hydrophilic antioxidants. Currently, most ORAC protocols use isolated compounds or freeze-dried fruit or vegetable samples. Here, the application of a standard hexane-type extraction method, which is more amenable to whole food carotenoid-containing samples, was investigated as a candidate extraction method for the ORAC assay. Variants of this method as well as of the standard ORAC extraction were compared for extraction efficiency. Finally, ORAC values were correlated with carotenoid content and shown to hold a loose negative correlation. Possible reasons for this are considered and discussed.

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Ji Tian, Ke-ting Li, Shi-ya Zhang, Jie Zhang, Ting-ting Song, Yong-jun Zhu, and Yun-cong Yao

Anthocyanins are protective pigments that accumulate in plant organs such as fruits and leaves, and are nutritionally valuable components of the human diet. The MYB10 transcription factor (TF) plays an important role in regulating anthocyanin biosynthesis in Malus crabapple leaves. However, little is known about how the promoter regulates McMYB10 expression and influences the substantial variation in leaf anthocyanin accumulation and coloration that is observed in different crabapple cultivars. In this study, we analyzed leaf coloration, anthocyanin levels, and the expression levels of McMYB10 in the leaves of 15 crabapple cultivars with three leaf colors at various development stages, and showed that the expression of McMYB10 correlates positively with anthocyanin accumulation. We also examined the relationship between the number of R6 and R1 elements in the McMYB10 promoters of the different cultivars and the pigmentation of the new buds of spring-red cultivars, as well as the methylation level of the McMYB10 promoters at different development stages in three representative crabapple cultivars. The ratio of R6/R1 minisatellites in the promoters correlated with the color and anthocyanin accumulation in new crabapple buds, and we concluded that the differences in promoter structure and methylation level of the McMYB10 promoters coordinately affect the leaf color of crabapple cultivars.

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John A. Juvik

Extensive epidemiological evidence suggests that carotenoids (including vitamin A), ascorbate (vitamin C), tocols (including vitamin E), and glucosinolate breakdown products exert anticarcinogenic effects in a range of human tissues. Consumption of fresh and processed vegetables with enhanced levels of these phytochemicals could reduce human risk of cancer. The vitamins play a major role as antioxidants, offering protection against cancer by preventing or reversing oxidative damage to DNA and other cellular components. Cruciferous vegetables contain glucosinolates (GSs), which, during mastication, are hydrolyzed by the enzyme myrosinase into bioactive breakdown products (BBPs), including sulforaphane. BBPs appear to induce synthesis of drug metabolism enzymes resulting in increased detoxification rates of carcinogens. This paper describes an interdisciplinary investigation designed to develop vegetable cultivars that offer chemoprotection from cancer at doses commensurate with a normal American diet. Initial work has focused on surveying sweet corn and Brassicae oleraceae germplasm for variation in vitamin and glucosinolate content in conjunction with in vitro and in vivo bioassays to determine which compounds and concentrations optimize chemoprotectant activity. Segregating populations from crosses between sweet corn and Brassica lines that vary in vitamin and GS concentrations will be assayed for chemical content and chemoprotectant activity, and genetically characterized using DNA marker technology to identify and map genes controlling these traits. This information will improve selection methodology in a breeding program aimed to develop brassica and sweet corn germplasm with enhanced cancer chemoprevention.

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Manette Schönfeld and Cary A. Mitchell

CowPea (Vigna unguiculata (L.) Walp.) is a candidate species for inclusion in a space-deployed Controlled Ecological Life Support System (CELSS) because it contributes to a balanced diet with its moderate protein content, high complex carbohydrate content, and low fat content, and because leaves and unripe pods as well as dry seeds are edible. Pour harvest scenarios were compared in the experimental line IT84S-2246 under controlled conditions with and without CO2 enrichment. Plants kept vegetative by removal of flowers and periodically stripped of fully expanded leaves yielded as much as either mixed-harvest scenario in which leaves were stripped at either 1- or 2-week intervals until pods started forming. The 2-week harvest scenario outyielded the 1-week scenario by 15 to 25%. The seed-only control produced the same amount of seeds as the 2-week leaf harvest scenario, but had lower total edible biomass because leaves were not harvested. Under 1000 ppm CO2, all treatments yielded from 30 to 70% more edible biomass than under non-CO2-enriched conditions. Research sponsored by NASA Cooperative Agreement NCC 2-100.